US 20150309021A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0309021 A1 Birnbaum et al. (43) Pub. Date: Oct. 29, 2015

(54) ADVANCED DRUG DEVELOPMENT AND tion No. 10/621,825, filed on Jul. 16, 2003, now Pat. MANUFACTURING No. 6,858,148, which is a continuation-in-part of application No. 10/206,524, filed on Jul. 25, 2002, now (71) Applicant: XRpro Sciences, Inc., Cambridge, MA abandoned, which is a continuation-in-part of applica (US) tion No. 09/859,701, filed on May 16, 2001, now Pat. No. 7,858,385. (72) Inventors: Eva R. Birnbaum, Los Alamos, NM (US); Andrew T. Koppisch, Flagstaff, (60) Provisional application No. 60/850,594, filed on Oct. AZ (US); Sharon M. Baldwin, Santa Fe, 10, 2006. NM (US); Benjamin P. Warner, Los Alamos, NM (US); T. Mark Publication Classification McCleskey, Los Alamos, NM (US); Jennifer A. Berger, Los Alamos, NM (51) Int. C. (US); Jeffrey J. Stewart, Cary, NC GOIN33/566 (2006.01) (US); Michael N. Harris, Los Alamos, (52) U.S. C. NM (US); Anthony K. Burrell, CPC ...... G0IN33/566 (2013.01); G0IN 2500/04 Naperville, IL (US) (2013.01) (57) ABSTRACT (21) Appl. No.: 14/693,094 X-ray fluorescence (XRF) spectrometry has been used for Filed: Apr. 22, 2015 detecting binding events and measuring binding selectivities (22) between chemicals and receptors. XRF may also be used for estimating the therapeutic index of a chemical. Forestimating Related U.S. Application Data the binding selectivities of a chemical versus chemical ana (60) Division of application No. 1 1/974,156, filed on Oct. logs, for measuring post translational modification of pro 10, 2007, which is a continuation-in-part of applica teins, and for drug manufacturing. US 2015/0309021 A1 Oct. 29, 2015

ADVANCED DRUG DEVELOPMENT AND These residues can form weak noncovalent bonds with other MANUFACTURING molecules. An effective binding between the protein, one example of a group of materials herein referred to as “recep RELATED APPLICATIONS tors', and the material that binds to the receptor, referred to 0001. This application is a Continuation of U.S. patent herein as “chemical, generally requires that many weak application Ser. No. 1 1/974,156, filed Oct. 10, 2007, which is bonds form between the protein receptor and the chemical. a continuation-in-part of U. S. patent application Ser. No. Chemicals include organic molecules, inorganic molecules, 09/859,701, filed May 16, 2001, now U.S. Pat. No. 7,858,385 salts, metal ions, and the like. The bonds between the protein issued Dec. 28, 2010, and a continuation-in-part of U.S. and the chemical form at the “binding site' of the protein. The patent application Ser. No. 10/206,524, filed Jul. 25, 2002, binding site is usually a cavity in the protein that is formed by and a continuation-in-part of U.S. patent application Ser. No. a specific arrangement of amino acids that often belong to 10/621,825 filed Jul 16, 2003, now U.S. Pat. No. 6,858,148 widely separated regions of the polypeptide chain and repre issued Feb. 22, 2005, all hereby incorporated by reference sent only a minor fraction of the total number of amino acids herein. This application also claims the benefit of U.S. Pro present in the chain. Chemicals should fit precisely into the visional Application Ser. No. 60/850,594 filed Oct. 10, 2006, binding site for effective binding to occur. The shape of these hereby incorporated by reference herein. binding sites can differ greatly among different proteins, and even among different conformations of the same protein. STATEMENT REGARDING FEDERAL RIGHTS Even slightly different conformations of the same protein may differ greatly in their binding abilities. For further dis 0002 This invention was made with government support cussion of the structure and function of proteins, see: Bruce under Contract No. DE-AC52-06NA25396 awarded by the Alberts et al., “Molecular Biology of the Cell", 2" edition, U.S. Department of Energy. The government has certain Garland Publishing, Inc., New York, 1989; and H. Lodish et rights in the invention. al., “Molecular Cell Biology', 4' edition, W. H. Freeman and Company, 2000. FIELD OF THE INVENTION 0007. After a receptor array is prepared, it is screened to 0003. The present invention relates generally to detecting determine which members have the desirable property or binding events and more particularly to estimating binding properties. U.S. Pat. No. 5,143,854 to M. C. Pirrung et al. selectivities for chemicals, analogs, and drugs being tested entitled “Large Scale Photolithographic Solid Phase Synthe with receptors and then manufacturing those having a high sis of Polypeptides and Receptor Binding Screening binding selectivity to the receptors. Thereof, which issued Sep. 1, 1992, hereby incorporated by reference, describes one Such screening method. A polypep BACKGROUND OF THE INVENTION tide array is exposed to a ligand (an example of a chemical) to 0004. The desire to hasten the identification of potentially determine which members of the array bind to the ligand. The important drugs, catalysts, chemicals and biological sensors, ligands described are radioactive, or are "tagged', i.e. medical diagnostics, and other materials is a constant chal attached via one or more chemical bonds to a chemical por lenge that has prompted the use of combinatorial synthetic tion that fluoresces when exposed to non-ionizing, ultraviolet screening strategies for synthesizing these materials and radiation. Thus, the attached portion, i.e. the tag, makes the screening them for desirable properties. Combinatorial Syn chemical visible by interrogation with ultraviolet radiation. thesis involves assembling a “library’, i.e. a very large num Tagged molecules have also been used to aid in sequencing ber of chemically related compounds and mixtures, usually in immobilized polypeptides as described, for example, in U.S. the form of an array on a Substrate Surface. High throughput Pat. No. 5,902,723 to W. J. Dower et al. entitled “Analysis of screening of an array involves identifying which members of Surface Immobilized Polymers Utilizing Microfluorescence the array, if any, have the desirable property or properties. The Detection.” which issued May 11, 1999. Immobilized array form facilitates the identification of a particular material polypeptides are exposed to molecules labeled with fluores on the Substrate. Combinatorial arrays and high-throughput cent tags. The tagged molecules bind to the terminal mono screening techniques have been used to solve a variety of mer of a polypeptide, which is then cleaved and its identity problems related to the development of biological materials determined. The process is repeated to determine the com Such as proteins and DNA because the screening techniques plete sequence of the polypeptide. can be used to rapidly assay many biological materials. 0008. It is generally assumed that the attachment of a 0005. The binding properties of a protein largely depend fluorescent tag to a chemical only serves to make visible the on the exposed Surface amino acid residues of its polypeptide otherwise invisible chemical, and does not alter its binding chain (see, for example, Bruce Alberts et al., “Molecular properties. Since it is well known that even Small changes to Biology of the Cell", 2" edition, Garland Publishing, Inc., the structure of a molecule could affect its function, this New York, 1989; and H. Lodish et al., “Molecular Cell Biol assumption that a tagged chemical, i.e. a 'surrogate', has the ogy”, 4" edition, W. H. Freeman and Company, 2000). These same binding affinity as the untagged chemical may not be a amino acid residues can form weak noncovalent bonds with valid one. Small structural changes that accompany even a ions and molecules. Effective binding generally requires the conformational change of a receptor have been known to formation of many weak bonds at a “binding site, which is affect the binding affinity of the receptor. The tagged surro usually a cavity in the protein formed by a specific arrange gates are structurally different from their untagged counter ment of amino acids. There should be a precise fit with the parts, and these structural differences could affect their bind binding site for effective binding to occur. ing affinities. Since binding affinities derived using tagged 0006. The chemical properties and in particular, the bind Surrogates are suspect, the binding properties of receptors and ing properties of a protein depend almost entirely on the chemicals should be evaluated using the untagged chemical exposed Surfaceamino acid residues of the polypeptide chain. or receptor and not with a tagged Surrogate. US 2015/0309021 A1 Oct. 29, 2015

0009 Pharmaceutical chemicals are the active ingredients Using Red and Green Fluorescent Proteins.” Analytical Bio in drugs, and it is believed that their therapeutic properties are chemistry, vol. 306, pp. 50-54, (2002); G. Y. J. Chen et al., linked to their ability to bind to one or more binding sites. The “Developing a Strategy for Activity-Based Detection of shapes of these binding sites may differ greatly among dif Enzymes in a Protein Microarray.” ChemBioChem, pp. 336 ferent proteins, and even among different conformations of 339, (2003); K. Martin et al., “Quantitative analysis of protein the same protein. Even slightly different conformations of the phosphorylation status and protein kinase activity on same protein may differ greatly in their binding abilities. For microarrays using a novel fluorescent phosphorylation sensor these reasons, it is extremely difficult to predict which chemi dye.” Proteomics, Vol. 3, pp. 1244-1255, (2003); J. Burbaum cals will bind effectively to proteins. and G. M. Tobal, “Proteomics in drug discovery. Current 0010 Development and manufacturing for a new pharma Opinion in Chemical Biology, Vol. 6, pp. 427-433. (2002); I. ceutical chemical for a drug, i.e. drug development, generally A. Hemmilä and P. Hurskainen, “Novel detection strategies involves determining the binding affinities between a poten for drug discovery. Drug Discovery Today, Vol. 7, pp. S150 tial pharmaceutical chemical (preferably a water soluble S156, (2002), Pages S150-S156; all incorporated herein by organic chemical that can dissolve into the blood stream) and reference. a receptor (generally a biological material Such as an enzyme 0015. Some screening methods are described in the fol or non-enzyme protein, DNA, RNA, human cell, plant cell, lowing patents, all of which are hereby incorporated by ref animal cell, and the like) at many stages of the drug develop CCC. ment process. The receptor may also be a microorganism (e.g. prion, virus, bacterium, spores, and the like) in whole or in 0016 U.S. Pat. No. 6,147,344 to D. Allen Annis et al. part. The drug development process typically involves proce entitled “Method for Identifying Compounds in a Chemical dures for combining potential pharmaceutical chemicals with Mixture', which issued Nov. 14, 2000, describes a method for receptors, detecting chemical binding between the potential automatically analyzing mass spectrographic data from mix pharmaceutical chemicals and the receptors and determining tures of chemical compounds. the binding affinity and kinetics of binding of a receptor to a 0017 U.S. Pat. No. 6,344,334 to Jonathan A. Ellman et al. chemical to form a complex or the kinetics of release of a entitled “Pharmacophore Recombination for the Identifica bound chemical from a complex. The binding affinity is tion of Small Molecule Drug Lead Compounds,” which defined herein as the associative equilibrium constant Ka for issued Feb. 5, 2002, describes a method for identifying a drug the following equilibrium expression: lead compound by contacting target biological molecules m(chemical)+receptor chemical-receptor with cross-linked binding fragments. complex (0018 U.S. Pat. No. 6,395,169 to Ole Hindsgaul et al. 0011 The binding affinity, Ka, is defined by equation (1) entitled "Apparatus for Screening Compound Libraries.” below. which issued May 28, 2002, describes an apparatus that employs frontal chromatography combined with mass spec Ka=chemical-receptor complex receptorchemi trometry to identify and rank members of a library that bind to cal” (1) a target receptor. 0012. In equation (1), chemical-receptor complex is the 0019. Other current high-throughput screening methods, concentration in moles per liter of the chemical-receptor along with their associated drawbacks, are listed in TABLE 1 complex, receptor is the concentration in moles per liter of below, which is taken from the following paper. H. Zhu and the receptor, m is the number of molecules of chemical that M. Snyder, “Protein Chip Technology (Review). Current bind to each molecule of receptor, and chemical is the Opinion in Chemical Biology, Vol. 7, pp.55-63, (2003). concentration in moles per liter of the chemical. Any effects due to concentration can be simplified if the concentration of TABLE 1 chemical used were the same for all receptors. 0013 Nowadays, the drug development process may Data involve the rapid screening of hundreds or thousands of Detection Technique Probe Labeling Acquirement chemicals in order to identify a “lead compound,” which is ELISA (Enzyme-Linked Enzyme-Linked Antibodies CCD one of the many tested that binds very strongly, i.e. has a high Immunosorbent Assay) Imaging Isotopic Labeling Radioisotope Labeled X-Ray binding affinity, with a particular receptor. After Such a lead Analyte Film Or compound has been identified, then other potential pharma Phospoimager ceutical chemicals similar in structure to the lead compound, Sandwich Immunoassay Fluorescently Labeled Laser which are referred to herein as “analogs” of the lead com Antibodies Scanning Surface Plasmon Resonance Receptor must be attached Refractive pound, are synthesized and tested in order to determine which to a surface Index Change of these chemicals, if any, exhibits an even higher binding Non-Contact Atomic Force None Surface affinity. Microscopy Topological Change 0014. The preparation and high-throughput Screening of Planar Waveguide Fluorescently Labeled CCD biological arrays is exemplified by the following papers: H. Antibodies Imaging Zhu and M. Snyder, “Protein Chip Technology (Review). SELDI (Surface Enhanced None Mass Current Opinion in Chemical Biology, Vol. 7, pp. 55-63, Laser Desorption Ionization Spectrometry Mass Spectrometry) (2003); G. MacBeath and S. L. Schreiber, “Printing Proteins Electrochemical Metal-Coupled Analyte Conductivity As Microarrays For High-Throughput Function Determina Measurement tion.” Science, vol. 289, pp. 1760-1763, (2000); E.T. Funget al., “Protein biochips for differential profiling.” Analytical Biotechnology, Vol. 12, pp. 65-69, (2001); T. Kukar et al., 0020 TABLE 1 shows that most of the listed screening “Protein Microarrays to Detect Protein-Protein Interactions methods have the same drawback: they require either radio US 2015/0309021 A1 Oct. 29, 2015

labeled chemicals, chemicals that have been altered with a 0025. Effective drugs are selective drugs; they bind to a fluorescent tag, or chemicals that have been altered with a specific desired receptor, bypassing other receptors, to pro metal tag. duce a desired therapeutic effect. This way, they target a 0021 X-ray fluorescence (XRF) spectrometry is a power specific disease in the body with minimal side effects; selec ful spectroscopic technique that has been used to determine tivity provides efficacy without undesirable side effects. the elements that are present in a chemical sample, and to 0026. The therapeutic index is a measure of drug selectiv determine the quantity of those elements in the sample. The ity that is ordinarily calculated from data obtained from underlying physical principle of the method is that when an experiments with animals. The therapeutic index of the drug atom of a particular element is irradiated with X-ray radia is typically defined as the ratio of two numbers, LD50/ED50, tion, the atom ejects a core electron Such as a Kshell electron. where LD50 is the dose of a drug that is found to be lethal (i.e. The resulting atom is in then an excited State, and it can return toxic) for 50 percent of the population of animals used for to the ground State by replacing the ejected electron with an testing the drug, and ED50 is the dose of the drug that is found electron from a higher energy orbital. This is accompanied by to have a therapeutic effect for 50 percent of that population. the emission of a photon, i.e. X-ray fluorescence, and the More broadly, it is a measure of the how much of the drug is photon energy is equal to the difference in the energies of the needed to produce a harmful effect relative to the amount two electrons. Each element has a characteristic set of orbital needed to produce a beneficial effect. Theratio LD50/ED50 is energies and therefore, a characteristic X-ray fluorescence therefore, a measure of the approximate “safety factor' for a (XRF) spectrum. drug; a drug with a high therapeutic index can presumably be 0022. An XRF spectrometer is an apparatus capable of administered with greater safety than one with a low index. irradiating a sample with an X-ray beam, detecting the X-ray 0027 Estimating the therapeutic index of a chemical fluorescence from an element included in the sample, and involves measuring the binding affinity of a chemical to a first using the X-ray fluorescence to determine which elements are receptor, and measuring the binding affinity of the same present in the sample and providing the quantity of these chemical to a second receptor. After measuring these binding elements. A typical, commercially available X-ray fluores affinities, the ratio of the binding affinity of the chemical cence spectrometer is the EDAX Eagle XPL energy disper divided by the amount of first receptor versus the binding sive X-ray fluorescence spectrometer, equipped with a micro affinity of the chemical divided by the amount of the second focus X-ray tube, lithium drifted silicon solid-state detector, receptor is determined. This ratio is provides an estimate of processing electronics, and Vendor Supplied operating soft the “therapeutic index. For an example of using DNA arrays Ware. In principle, any element may be detected and quanti with optical fluorescence high-throughput screening to esti fied with XRF. mate a therapeutic index, see for example, R. Ulrich and S.H. 0023 U. S. Patent Application 20030027129 entitled Friend, “Toxicogenomics and Drug Discovery: Will New “Method for Detecting Binding Events. Using Micro-X-ray Technologies Help Us Produce Better Drugs?.” Nature Fluorescence Spectrometry, incorporated by reference Reviews Drug Discovery, v.1, pp. 84-88 (2002), incorporated herein by reference. herein, describes a method for detecting binding events using 0028. There remains a need for simpler methods for mea micro-X-ray fluorescence spectrometry. According to the Suring binding affinities and selectivities, estimating the 129 patent application, receptors are exposed to at least one therapeutic index of a chemical, and for expediting drug potential chemical and arrayed on a substrate Support. Each manufacturing, member of the array is exposed to X-ray radiation. The mag 0029. Therefore, an object of the present invention is to nitude of a detectable X-ray fluorescence signal for at least one provide a method for measuring binding affinities of chemi element can be used to determine whether a binding event cals. between a chemical and a receptor has occurred, and can 0030. Another object of the present invention is to provide provide information related to the extent of binding between a method for measuring selectivity of chemicals binding to the chemical and receptor. receptors. 0024 U. S. Patent Application 2004.0017884 entitled “Flow Method And Apparatus For Screening Chemicals 0031 Yet another object of the invention is to provide a Using Micro X-Ray Fluorescence,” which was published on method for estimating the therapeutic index of a chemical. Jan. 29, 2004, incorporated by reference herein, describes a 0032 Still another object of the invention is to provide a method for identifying binding events between potential method that employs X-ray fluorescence for drug manufac pharmaceutical chemicals and receptors (e.g. proteins). The ture. method involves modifying a mixture of potential pharma 0033. Additional objects, advantages and novel features of ceutical chemicals by adding at least one receptor to the the invention will be set forth in part in the description which mixture. After allowing sufficient time for any bound com follows, and in part will become apparent to those skilled in plex between any of the potential pharmaceutical chemicals the art upon examination of the following or may be learned and any of the receptors to form, if such a complex can form, by practice of the invention. The objects and advantages of the the resulting solution is flow separated into at least two com invention may be realized and attained by means of the instru ponents. Each component is exposed to an X-ray excitation mentalities and combinations particularly pointed out in the beam. If the exposed component emits a detectable X-ray appended claims. fluorescence signal, that component is isolated. The identity of any isolated component can be determined using one or SUMMARY OF THE INVENTION more standard analytical techniques, such as gas chromatog 0034. In accordance with the purposes of the present raphy, liquid chromatography, mass spectrometry, nuclear invention, as embodied and broadly described herein, the magnetic resonance spectroscopy, infrared spectroscopy, present invention includes a method for estimating the bind ultraviolet spectroscopy, visible spectroscopy, elemental ing selectivity of a chemical having at least one heavy element analysis, cell culturing, immunoassaying, and the like. (i.e. an element having an atomic number of nine or higher) to US 2015/0309021 A1 Oct. 29, 2015

at least two receptors. The method includes establishing a from the measured X-ray fluorescence signal of the of the first baseline X-ray fluorescence signal for a heavy element in a analog-receptor complex; calculating the net X-ray fluores portion of a first receptor and in a portion of at least one more cence signal due to the second heavy element present in the receptor that may be the same or different from the first second analog-receptor complex by Subtracting the baseline receptor, the heavy element being present in a chemical to be X-ray fluorescence signal of the second portion of the second tested for binding to the receptors; exposing the receptors to receptor from the measured X-ray fluorescence signal of the the chemical and allowing the chemical to bind to them to second analog-receptor complex; estimating the selectivity of form a first chemical-receptor complex and at least one more the chemical to binding to the receptors by dividing the net chemical-receptor complex; measuring the X-ray fluores X-ray fluorescence of the first chemical-receptor complex by cence signals due to the heavy element in the first chemical the amount of receptor in the first portion of the first receptor receptor complex and in the at least one more chemical to obtaina first quotient, dividing the net X-ray fluorescence of receptor complex; Subtracting the baseline X-ray fluorescence the second chemical-receptor complex by the amount of signal of the first receptor from the measured X-ray fluores receptor in the first portion of the second receptor to obtain a cence signal of the of the first chemical-receptor complex to second quotient; estimating the selectivity of the analog to obtain a first net X-ray fluorescence signal; Subtracting the binding to the receptors by dividing the net X-ray fluorescence baseline X-ray fluorescence signal of the at least one more of the first analog-receptor complex by the amount of receptor receptor from the measured X-ray fluorescence signal of the in the second portion of the first receptor to obtain a third of the at least one more chemical-receptor complex to obtain quotient, dividing the net X-ray fluorescence of the second at least one more net X-ray fluorescence signal; and then analog-receptor complex by the amount of receptor in the estimating the selectivity of the chemical by dividing the first second portion of the second receptor to obtain a fourth quo net X-ray fluorescence signal by the amount of receptor in the tient; and comparing first quotient to the second quotient to portion of the first receptor to obtain a first quotient, dividing the third quotient to the fourth quotient. the at least one more net X-ray fluorescence signal by the 0036. The invention also includes a method for manufac amount of receptor in the portion of the at least one more turing a drug. The method includes establishing a baseline receptor to obtain at least one more quotient, and then com X-ray fluorescence signal for a heavy element in a portion of paring the first quotient to the at least one more quotient. a first receptor and in a portion of at least one more receptor 0035. The invention also includes a method forestimating that may be the same or different from the first receptor, the the binding selectivity of a chemical versus at least one analog heavy element being present in a chemical to be tested for of the chemical to at least two receptors, the chemical and the binding to the receptors; exposing the receptors to the chemi at least one analog each having at least one heavy element. cal and allowing the chemical to bind to them to form a first The method includes establishing a baseline x-ray fluores chemical-receptor complex and at least one more chemical cence signal for a first heavy element in a first portion of a first receptor complex; measuring the X-ray fluorescence signals receptor and in a first portion of a second receptor, the first due to the heavy element in the first chemical-receptor com heavy element being present in a chemical to be tested for plex and in the at least one more chemical-receptor complex; binding to the receptors; establishing a baseline X-ray fluo Subtracting the baseline X-ray fluorescence signal of the first rescence signal for a second heavy element in a second por receptor from the measured X-ray fluorescence signal of the tion of the first receptor and in a second portion of a second of the first chemical-receptor complex to obtain a first net receptor, the second heavy element being present in an analog X-ray fluorescence signal; Subtracting the baseline X-ray fluo of the chemical to be tested for binding to the receptors: rescence signal of the at least one more receptor from the exposing the first portions of the receptors to the chemical and measured X-ray fluorescence signal of the of the at least one allowing the chemical to bind to them to forma first chemical more chemical-receptor complex to obtain at least one more receptor complex and a second chemical-receptor complex; net X-ray fluorescence signal; estimating the selectivity of the measuring the X-ray fluorescence signal due to the first heavy chemical by dividing the first net X-ray fluorescence signal by element present in the first chemical-receptor complex and in the amount of receptor in the portion of the first receptor to the second chemical-receptor complex; exposing the second obtain a first quotient, dividing the at least one more net X-ray portions of the receptors to the analog and allowing the analog fluorescence signal by the amount of receptor in the portion of to bind to them to form a first analog-receptor complex and a the at least one more receptor to obtain at least one more second analog-receptor complex; measuring the X-ray fluo quotient, and then comparing the first quotient to the at least rescence signal due to the second heavy element present in the one more quotient; and manufacturing the chemical in Suffi first analog-receptor complex and in the second analog-re cient quantity for use as a drug if the first quotient and the at ceptor complex; calculating the net X-ray fluorescence signal least one more quotient are different by at least one percent. due to the first heavy element in the first chemical-receptor 0037. The invention also includes a method for manufac complex by Subtracting the baseline X-ray fluorescence signal turing a drug. The method includes establishing a baseline of the first portion of the first receptor from the measured X-ray fluorescence signal for a first heavy element in a first X-ray fluorescence signal of the of the first chemical-receptor portion of a first receptor and in a first portion of a second complex; calculating the net X-ray fluorescence signal due to receptor, the first heavy element being present in a chemical the first heavy element present in the second chemical-recep to be tested for binding to the receptors; establishing a base tor complex by Subtracting the baseline X-ray fluorescence line X-ray fluorescence signal for a second heavy element in signal of the first portion of the second receptor from the a second portion of the first receptor and in a second portion measured X-ray fluorescence signal of the second chemical of a second receptor, the second heavy element being present receptor complex, calculating the net X-ray fluorescence sig in an analog of the chemical to be tested for binding to the nal due to the second heavy element in the first analog receptors; exposing the first portions of the receptors to the receptor complex by Subtracting the baseline X-ray chemical and allowing the chemical to bind to them to form a fluorescence signal of the second portion of the first receptor first chemical-receptor complex and a second chemical-re US 2015/0309021 A1 Oct. 29, 2015 ceptor complex; measuring the X-ray fluorescence signal due ing the net X-ray fluorescence signal due to the heavy element to the first heavy element present in the first chemical-recep in the first chemical-receptor complex by Subtracting the tor complex and in the second chemical-receptor complex; baseline X-ray fluorescence signal of the first portion of recep exposing the second portions of the receptors to the analog tor from the measured X-ray fluorescence signal of the first and allowing the analog to bind to them to form a first analog chemical-receptor complex; calculating the net X-ray fluores receptor complex and a second analog-receptor complex; cence signal due to the heavy element in the second chemical measuring the X-ray fluorescence signal due to the second receptor complex by Subtracting the baseline X-ray fluores heavy element present in the first analog-receptor complex cence signal of the separate portion of receptor from the and in the second analog-receptor complex; calculating the measured X-ray fluorescence signal of the second chemical net X-ray fluorescence signal due to the first heavy element in receptor complex; and estimating the binding selectivity of the first chemical-receptor complex by Subtracting the base the chemical in the first solution versus the second solution by line x-ray fluorescence signal of the first portion of the first dividing the net X-ray fluorescence signal of the first chemi receptor from the measured X-ray fluorescence signal of the cal-receptor complex by the amount of receptor in the first of the first chemical-receptor complex; calculating the net portion of the receptor to obtain a first quotient, dividing the X-ray fluorescence signal due to the first heavy element net X-ray fluorescence of the second chemical-receptor com present in the second chemical-receptor complex by Subtract plex by the amount of receptor in the separate portion of the ing the baseline X-ray fluorescence signal of the first portion receptor to obtain a second quotient, and then comparing the of the second receptor from the measured X-ray fluorescence first quotient to the second quotient. signal of the second chemical-receptor complex; calculating 0039. The invention also includes a method forestimating the net X-ray fluorescence signal due to the second heavy the relative effectiveness of at least two drugs, each drug element in the first analog-receptor complex by Subtracting having at least one heavy element. The method includes pro the baseline X-ray fluorescence signal of the second portion of viding a sample from a medical patient, the sample compris the first receptor from the measured X-ray fluorescence signal ing at least a first portion and a second portion of a first of the of the first analog-receptor complex, calculating the net receptor and a first portion and a second portion of a second X-ray fluorescence signal due to the second heavy element receptor; establishing a baseline X-ray fluorescence signal for present in the second analog-receptor complex by Subtracting a first heavy element in the first portion of the first receptor the baseline X-ray fluorescence signal of the second portion of and in the first portion of the second receptor, the first heavy the second receptor from the measured X-ray fluorescence element being present in a first drug to be tested for binding to signal of the second analog-receptor complex; estimating the the first and second receptors; establishing a baseline X-ray selectivity of the chemical to binding to the receptors by fluorescence signal for a second heavy element in the second dividing the net X-ray fluorescence of the first chemical portion of the first receptor and in the second portion of the receptor complex by the amount of receptor in the first portion second receptor, the second heavy element being present in a of the first receptor to obtain a first quotient, dividing the net second drug to be tested for binding to first and second recep X-ray fluorescence of the second chemical-receptor complex tors; exposing the first portion of the first receptor and the first by the amount of receptor in the first portion of the second portion of the second receptor to the first drug and allowing receptor to obtain a second quotient; estimating the selectivity the drug to bind to the first receptor to form a first drug of the analog to binding to the receptors by dividing the net receptor complex, and to bind to the second receptor to form X-ray fluorescence of the first analog-receptor complex by the a second drug-receptor complex, measuring the X-ray fluo amount of receptor in the second portion of the first receptor rescence signal due to the first heavy element present in the to obtain a third quotient, dividing the net X-ray fluorescence first drug-receptor complex and in the second drug-receptor of the second analog-receptor complex by the amount of complex; exposing the second portion of the first receptor and receptor in the second portion of the second receptor to obtain the second portion of the second receptor to the second drug a fourth quotient; and comparing first, second, third, and and allowing the second drug to bind to the first receptor to fourth quotients to determine if the chemical or analog is the form a third drug-receptor complex, and to bind to the second more selective and manufacturing the more selective one in receptor to form a fourth drug-receptor complex; measuring Sufficient quantity for use as a drug. the X-ray fluorescence signal due to the second heavy element 0038. The invention also includes a method for comparing present in the third drug-receptor complex and in the fourth the ability of at least one chemical in a first solution to bind to drug-receptor complex; calculating a first net X-ray fluores a portion of at least one receptor versus the ability of that cence signal due to the first heavy element present in the first chemical in a second solution to bind to a separate portion of drug-receptor complex by Subtracting the baseline X-ray fluo the same at least one receptor. The method includes establish rescence signal of the first portion of the first receptor from ing a baseline X-ray fluorescence signal for a heavy element the measured x-ray fluorescence signal of the of the first in a first portion of a receptor and for the heavy element in a drug-receptor complex; calculating a second net X-ray fluo separate portion of the receptor, the heavy element being rescence signal due to the first heavy element present in the present in a chemical that is being tested for binding to the second drug-receptor complex by Subtracting the baseline receptor, exposing the first portion of the receptor to a first X-ray fluorescence signal of the first portion of the second Solution that includes the chemical, and allowing the chemi receptor from the measured X-ray fluorescence signal of the cal to bind to the receptor to form a first chemical-receptor second drug-receptor complex; calculating a third net X-ray complex; exposing the separate portion of the receptor to a fluorescence signal due to the second heavy element in the second solution also includes the chemical, and allowing the third drug-receptor complex by Subtracting the baseline X-ray chemical to bind to the receptor to form a second chemical fluorescence signal of the second portion of the first receptor receptor complex; measuring the X-ray fluorescence signals from the measured x-ray fluorescence signal of the of the third due to the heavy element in the first chemical-receptor com drug-receptor complex; calculating a fourth net X-ray fluo plex and in the second chemical-receptor complex; calculat rescence signal due to the second heavy element present in the US 2015/0309021 A1 Oct. 29, 2015

fourth drug-receptor complex by Subtracting the baseline ratios. All of these methods may be used with multiple X-ray fluorescence signal of the second portion of the second samples to provide comparisons between samples, or with receptor from the measured X-ray fluorescence signal of the single samples to obtain information about the sample. fourth drug-receptor complex; calculating binding quotients for the first drug by dividing the first net X-ray fluorescence Substrates signal by the amount of receptor in the first portion of the first receptor to obtain a first quotient, dividing the second net 0043. A key improvement to US Patent Application X-ray fluorescence signal by the amount of receptor in the first 20040235059 is the use of sample substrates which minimize portion of the second receptor to obtain a second quotient; noise. Characteristics to be optimized include the thickness of calculating binding quotients for the second drug by dividing the substrate; the chemical and elemental composition of the the third net X-ray fluorescence signal by the amount of recep substrate; and the variability in the chemical composition of tor in the second portion of the first receptor to obtain a third the surface of the substrate. quotient, dividing the fourth net X-ray fluorescence signal by the amount of receptor in the second portion of the second 0044 Substrates are preferably thin enough to minimize receptor to obtain a fourth quotient; and comparing the first, the scattering of incident X-rays. Thick Substrates cause scat second, third, and fourth quotients to estimate the relative tering in the X-ray portion of the electromagnetic spectrum. effectiveness of the first drug versus the second drug. Preferably, the substrates are less than 500 microns thick, and 0040. The invention also includes a method forestimating more preferably the substrates are less than about 50 microns binding affinity. The method involves depositing a portion of thick. Most preferably, the substrates are between twenty (20) a receptor on a Substrate; establishing a baseline X-ray fluo nanometers thick and twenty-five (25) microns thick. rescence signal for a heavy element in the portion of the Examples of materials which may be conveniently used receptor, the heavy element being present in a chemical being include 0.02 micron thick, 99.99% pure gold foil, available testing for binding to the receptor, exposing the receptor to a from Lebow Company, 5960 Mandarin Ave., Goleta, Calif. Solution comprising the chemical at a first temperature, and 93.117 U.S.A.: 0.07 micron thick, 99.95% pure aluminum allowing the chemical to bind to the receptor to form a chemi foil, available from Lebow Company, 5960 Mandarin Ave., cal-receptor complex; measuring the X-ray fluorescence sig Goleta, Calif. 93117 U.S.A.: 0.1 micron through 20 micron nal due to the heavy element in the chemical-receptor com thick PARYLENE NR), available from Lebow Company, plex using excitation photons having an energy of at least 300 5960 Mandarin Ave., Goleta, Calif. 93117 U.S.A.; and 0.5 electron-volts to electronically excite the heavy element, and micron thick to 25.0 micron thick, 99.99% pure titanium foil, detecting emission photons using an X-ray detector having a available from Lebow Company, 5960 Mandarin Ave., dead time, the emission photons being generated from an Goleta, Calif. 93117 U.S.A.; and 4 micron thick to 8 micron excited state of the heavy element, the excited state of the thick polypropylene, available from Lebow Company, 5960 heavy element having a fluorescence lifetime that is less than Mandarin Ave., Goleta, Calif. 93117 U.S.A. Other substrates the dead time of the X-ray detector, calculating the net X-ray that are conveniently used are AP1, AP3, ProLINE Series 10, fluorescence signal due to the heavy element in the chemical ProLINE Series 20, DuraBeryllium substrates from Moxtek, receptor complex by Subtracting the baseline X-ray fluores 452 West 1260 North, Orem, Utah 84057. Other substrates cence signal of the portion of receptor from the measured that are conveniently used are UltraleneR), Mylar, polycar X-ray fluorescence signal of the chemical-receptor complex; bonate, Prolene, and Kapton, available from SPEX CertiPrep and estimating the binding affinity of the chemical for the Ltd, 2 Dalston Gardens, Stanmore, Middx HA7 1BQ. receptor by dividing the net X-ray fluorescence signal of the ENGLAND. Other substrates that may be conveniently used chemical-receptor complex by the amount of the receptor in are Hostaphan(R), polyester, and Etnom R available from Chemplex Industries, Inc., 2820 SW 42nd Avenue, Palm City, the portion of the receptor. Fla. 34990-5573 USA. The substrate is preferably a polymer, 0041. The invention also includes a method for detecting and more preferably is a polymer that contains carbon. Radia protein modification. The method includes establishing a tion produced by radioactive materials will produce noise in baseline X-ray fluorescence signal for a heavy element in a the measurement. The substrate is preferably substantially portion of a protein; exposing the portion of protein to con free of the elements technetium, thorium, and uranium. Most ditions that may alter the amount of the heavy element present of the chemicals to be studied using the present invention will in the portion of the protein and then measuring the X-ray contain one or more of the following elements: oxygen, fluo fluorescence signal due to the heavy element; and Subtracting rine, phosphorus, Sulfur, and chlorine. Therefore, to minimize the baseline X-ray fluorescence signal from the measured unwanted signal from these elements in the focusing chip, the X-ray fluorescence signal. focusing chip should consist of less than 50% by mass of at least one of these elements (fluorine, oxygen, phosphorus, DETAILED DESCRIPTION sulfur, and chlorine). The substrate preferably comprises less than about 50% of at least one of the elements selected from Overview the group comprising oxygen, fluorine, phosphorus, Sulfur, 0042. The present invention consists of improvements to and chlorine. Based on their structural and spectroscopic the invention described in published US Patent Application properties, elements that are useful in the window include 20040235059 “Drug Development and Manufacturing to beryllium, carbon, silicon, aluminum, iron, cobalt, and gold. Benjamin P. Warner, T. Mark McCleskey, and Anthony K. The substrate therefore preferably contains at least 10% of at Burrell, filed Jun. 29, 2004. These improvements consist of least one of the following elements: beryllium, carbon, sili the following additional steps to be used in the process that con, aluminum, iron, cobalt, and gold. The Substrate prefer improve the signal or signal to noise ratios obtained from the ably comprises greater than about 10% of at least one of the measurement, and specific requirements for preparation of elements selected from the group comprising beryllium, car samples that provide improved signals or signal to noise bon, silicon, aluminum, iron, cobalt, and gold. US 2015/0309021 A1 Oct. 29, 2015

0045. Substrates may absorb scattered X-rays that may Walker, II; who stated that: “Immobilized metal affinity contribute to noise in the measurement. Substrates that absorb chromatography (IMAC) is a powerful technique which scattered X-rays preferably have an X-ray emission peak is routinely used in the purification of biomolecules between about 2.984 KeV (e.g. silver) and about 6.947 KeV containing a hexa-histidine tag, as well as phosphory (e.g. erbium) to avoid overlap with signals of interest. X-ray lated peptides/proteins. This wide range of affinities is emission peaks that are at higher energies than about 6.947 afforded by the chelation of specific metal ions by an KeV are generally non-productive for exciting the sample if it immobilized poly-valent ligand (i.e. IDA-Ni(II) for contains Sulfur, phosphorus, or chlorine; although in cases hexa-His tags and IDA-Fe(III) for phosphorylated spe where the sample contains elements such as selenium, X-rays cies). The utility of IMAC surfaces formed from SAMs of higher energy may be useful. X-ray emission peaks from has been demonstrated with SPR. In this work, chelated the substrate that have energies between about 1.978 KeV metals allows for high affinity capture and optimal ori (e.g. iridium) and about 2.984 KeV (e.g. silver) are generally entation of tagged proteins for Subsequent protein/pro counter-productive if the sample contains Sulfur, chlorine, or tein interaction studies. We have developed a SAM phosphorus; this is the typical case for biological or pharma based biochip that takes full advantage of these ceutically-important samples. If the sample to be measured properties and allows for Volume containment, sample contains phosphorus, then the Substrate should not contain concentration and direct interface to and detection by Zirconium, phosphorus, platinum, gold, niobium, mercury, or MALDI-MS. Three distinct SAM regions were pat thallium. If the sample to be measured contains sulfur, then terned on a gold-coated biochip in a 96 well format. the Substrate should not contain thallium, molybdenum, Sul Each site was comprised of a circular capture Zone con fur, lead, bismuth, technetium, or ruthenium. If the sample to taining either NTA-Ni(II) or NTA-Fe(III) chemistry. be measured contains chlorine, then the substrate should not Each site is surrounded by a non-wettable, protein-re contain technetium, ruthenium, chlorine, rhodium, or palla sistant SAM region and is capable of retaining sample dium. X-ray emission peaks from Substrates that are less than volumes in excess of 25ul. Centered in the capture Zone 1.978 KeV are believed to be unproductive for exciting bio was a 0.6 mm analysis Zone consisting of a protein logical samples. The substrate should be substantially free of resistant SAM. After chelating the metal, samples con the elements that are not intended to be measured in the taining either His tags or phosphorylated amino acid sample if energy dispersive X-ray fluorescence (EDXRF) is to residues were applied and purified on-chip. Subsequent be used for analysis. Wavelength dispersive X-ray fluores release, concentration and detection of the captured ana cence (WDXRF) have been shown to differentiate different lytes was facilitated by focusing and the addition of chemical forms of elements, such as the Sulfur compounds matrix.’ sulfate and sulfite; if WDXRF is used, then the substrate should be substantially free of the same chemical form of the (0.048 “Effects of Nanoporous Silica Thin Film Param element to be measured in the sample. In this paragraph, eters on LDI-MS Performance' by Srinivas Iyer; Raea substantially free is defined as a substrate which produces less Hicks; Steven Madrid; and Andrew Dattelbaum; who than 10% of the signal produced by the sample, which when Stated that “The use of MALDI-TOF MS in Small mol ecule analysis has been limited due to matrix interfer measured using the same X-ray fluorescence conditions. ence. We have shown that ordered nanoporous silica thin Substrates: Focusing Chips films on silicon substrates allow for the analysis of small organic molecules without the addition of a matrix. Pat 0046. The invention also consists of preparing a protein terned nanoporous films are prepared by an evaporation sample on a focusing chip. Focusing chips typically consist of self-assembly method, followed by selective UV expo a Substrate having a pattern of hydrophobic and hydrophilic Sure, to prepare Surfaces that may be spotted with many regions. Preferably, each hydrophilic region is circumscribed different analytes. The patterned films have been shown by a hydrophobic region. When a sample of an aqueous to remain active for over a year under atmospheric con Solution is placed on a hydrophilic region of the chip, the ditions. Here we report the effect of several variables, Surrounding hydrophobic region repels the Solvent and including film thickness, film porosity, instrumental set causes the sample to dry preferentially over the hydrophilic tings and spotting technique, to optimize the mass spec region. The drying may be accomplished through any means, analysis of Small peptides and other biomolecular including simple ambient air drying or heating. In this way, samples on nanoporous silica thin films. Mesoporous larger droplet may be dried into a predefined region using a thin films are made by withdrawing an oZonated silicon focusing chip instead of a non-focusing slide. Focusing chips crystal at a steady rate from a solution of TEOS, water, have been developed for depositing proteins for Matrix ethanol, HCl, and templating Surfactant, then UV-pat Assisted Laser Desorption Ionization Mass Spectrometry terning the film to remove the template and expose ana (MALDI-MS). Some of these focusing chips were described lyte-absorbing pores. Film thickness and pore size can at the 54th ASMS Conference on Mass Spectrometry, May be controlled by withdrawal speed and choice of surfac 28-Jun. 1, 2006, Washington State Convention & Trade Cen tant, respectively. These films are mounted directly onto ter, Seattle, Wash., described on the internet at: http://www. a standard MALDI plate. Fims of varying thickness and asms.org/Desktopmodules/inmergeabstractsearch/program pore size were examined. Spotting conditions were also printView.as px?sess=WP13, and quoted below in the examined using a solution of FITC labeled angiotensin. bulleted list: Before MS, the spots were examined by fluorescent 0047. “Applications of a SAM-IMAC Biochip for the microscopy to determine spot spreading and coverage. Purification and Concentration of Bioengineered or MS analysis was conducted on a Voyager DE-STR in Phosphorylated Proteins with Detection by MALDI positive ion, reflector mode. Our data indicates that these MS” by Christopher M. Belisle: Irene Y. Chen; Julie K. films require no special storage conditions to remain Mirshad, Udo Roth; Kerstin Steinert; and John A. stable for over a year. Preliminary studies show that US 2015/0309021 A1 Oct. 29, 2015

mesoporous silica thin films yield acceptable results in and the sample desalted. Resulting mass spectra showed terms of intensity and resolution for Small molecules a clean digestion, affording a high score identification on such as peptides and C60 and its derivatives. Reliable proteomics search engine. Analysis on crude biological data was obtained for acid hydrolysates of bacterial sid samples like human plasma will be also presented. The erophores. Optimizing spotting methods improved sig different parameters, monolith porosity and thickness, nal intensity by two orders of magnitude while retaining nature of monomers, droplet handling, are currently resolution. Fluorescence microscopy data shows that under investigation. We are also testing other monolithic spotting Small amounts of analyte, followed by remov phases we also developed for nano-LC capillary column ing excess Solution after a minute gives a uniform spread for phopshopeptide (IMAC) and glycopeptide (phenyl of sample, this is further substantiated by MS data. With boronic acid) isolation.” peptides and Small molecules, we observe that thinner 0050 “The Use of Reactive Surfaces for On-Chip films perform better. Small changes in pore size do not Immobilization of Biomolecules and Subsequent Puri appear to affect desorption-ionization significantly. fication/Concentration of Analytes for MALDI-MS. by However, when compared with MALDI, higher laser Julie K Mirshad; Christopher M Belisle; Irene Y Chen: fluences are needed with these thin films. Future work Udo Roth; Kerstin Steinert; and John A Walker who will be directed towards improving sensitivity of this stated that Currently in proteomics there is an increasing approach.” demand for high affinity purification of small amounts of 0049. “Functionalized monolithic surfaces for single analytes from complex samples. This requires methods droplet sample handling by Mohammed Kaijout; Xiay that minimize sample handling and analyte loss while ang Wang: Christian Rolando: Séverine Le Gac; who being amenable to high-throughput processing. Typi stated that “We describe here functionalized monolithic cally, highly selective purifications have been achieved Surfaces for single droplethandling. In many cases mass by immobilized biomolecules such as mAb, steptavidin/ spectrometry analysts must face to single droplet han biotin or nucleic acids. The most frequently used chem dling either as the initial sample input of the analysis or istry reported in literature for such immobilizations are for sample preparation before nano-ESI or MALDI. The active esters (i.e. N-Hydroxysuccinimide-NHS). We conventional strategy consists of dissolving the droplet have developed a novel biochip platform used in in more solvent so as to perform sample preparation on MALDI-MS that consists of multiple sites with differing devices working with continuous liquid flow. At the final Zones of wettability which enable immobilization of stage liquid must be concentrated to go back to a the these biomolecules with subsequent concentration of droplet state. Therefore the development of surface the captured analytes. Three distinct SAM regions were chemistry for the purification of single liquid droplets patterned on a gold-coated biochip in a 96 well format. has a strong interest in order to minimize sample han Each site was comprised of a circular capture Zone con dling, contamination and loss. At the routine sensitivity taining the active NHS chemistry. These sites are level of mass spectrometry, monolayer Surface chemis capable of retaining sample Volumes in excess of 25 ul try does not provide enough interaction sites for real and are surrounded by a non-wettable, protein-resistant world analysis. Subsequently we decided to use mono background SAM region. Centered in the capture Zone lithic polymer chemistry that we had previously devel was a 0.6 mm analysis Zone consisting of a highly wet oped for capillary columns dedicated to proteomics table, protein-resistant SAM that facilitates MALDI per applications and that affords 1 micron-thick film. Meth formance. After protein immobilization, samples con acrylate-based monoliths were anchored on a gold Sur taining complementary antigens in complex mixtures face using a linker bearing at its two extremities a dis were applied and purified on-chip. Subsequent release, ulfide moiety and a methacrylate group respectively, and concentration and detection of the captured analytes polymerisation was done according to an original living were facilitated by focusing and the addition of matrix. method. Non reactive and hydrophobic butylmethacry In initial experiments, analytes were used in spike/re late (BMA) or reactive glycidylmethacrylate (GMA) covery experiments with various protein digests to test may be used and mixed together in any proportions. for specificity and the limits of detection (LOD) for the Either silicon wafer coated with gold or low-cost printed biochip. In Subsequent experiments, mAb were immo board circuit coated with copper or gold may be used as bilized and proteins were purified on-chip and then the starting Surface. Using printed board enables rapid digested in situ with trypsin. Sample clean-up was per prototyping of patterned surfaces. Most of the experi formed by either transferring to a reverse-phase 3 Zone ments described were done either on plain gold Surface biochip or by SPE. Results comparing the advantages of or 400 um gold spots. We first tested the desalting effi this technique are discussed.” ciency using a laurylmethacrylate phase. One microliter 0051. “Enhanced MALDI ionization efficiency at the droplets of a salted (20%) solution containing various metal-matrix interface: practical and mechanistic con peptides (1 picomolar) were placed on the Surface, and sequences of samplethickness and preparation method.” interactions were forced by several aspiration/ejection by Gregor McCombie; and Richard Knochenmuss, who cycles. Spots were washed with water, extracted with stated that "Electrosprayed spots of varying thickness acetonitrile/water and analyzed offline either by ESI or were evaluated for use as homogenous, high efficiency MALDI. In both cases, no adducts were detected on the MALDI samples. The layer at the metal/matrix interface mass spectra. The second test involved a functionalized was found to give exceptionally strong signals. Ion glycidylmethacrylate on which was first linked trypsin. yields and intensity ratios can be understood in the con After washing of the surface, one microliter of Cyto text of the previously described quantitative MALDI chrom C (1 picomolar) was deposited and let on the model including the matrix-metal interfacial ionization surface for a quarter of an hour. More water was added potential reduction effect (Knochenmuss, R. Anal. US 2015/0309021 A1 Oct. 29, 2015

Chem. 2004, 76,3179-3184). The absolute and relative erogeneous MALDI sample deposit requires searching stabilities of ion signals were found to be at least a factor for “sweet spot” to obtain the optimum mass spectrum of 2 better for the thin electrosprayed spots, compared to with a stainless steel substrate. One of the greatest spots prepared by dried droplet methods. The sample advantages of the untreated silicon Substrate is that it preparation method was used for the analysis of yeast leads to a homogeneous sample spot so that a search for lipid composition. In order to detect differences in lipid a Sweet spot is not necessary. Comparison of Small mol content a stable sample preparation is necessary. Experi ecule LDI analysis of riboflavin showed significant dif ments were performed on a commercial MALDITOF ference between silicon and stainless steel substrates. instrument (Voyager DE STR, Applied Biosystems The stainless steel target gave Small analyte peaks only Framingham, Mass.). The mass range was from m/Z 100 at high laser energy that also produced extensive back to 1500. The electrospray sample deposition was done ground due to ablation of material on the metal target. In on a home built electrospray device using a CTC Pal contrast, the untreated silicon Substrate showed intense autosampler for the sample injection. The spray Voltage analyte peaks without background noise even at full was 5 kV and the distance of the needle to the sample laser energy. It was difficult to obtain signal from the plate 6 mm. Lipid extracts from yeast were done using stainless steel target, but highly reproducible mass spec the Bligh and Dyer method. Electrospary deposition of tra could be obtained for non-porous silicon substrate. matrix (DHB) and analyte (Substance P or reserpine) on BSA, Escherichia coli and phosphorylase b MALDI a MALDI target plate of varying spray times allowed us mass spectra could also be obtained easily on the silicon to control the thickness of the sample. Spray times substrate with high spot-to-spot reproducibility. The between 2 and 30 seconds were used to demonstrate that silicon target required somewhat higher threshold the MALDI ion yield is 10 to 100 times higher at the energy compared to the metal target, but showed some metal/matrix interface. The results were consistent with, what better signal-to-noise ratio and resolution than and could be explained by a previously described stainless steel substrate for small molecule as well as MALDI ionization model (Knochenmuss, R. Anal. high molecule analyses. A Surface characteristic of non Chem. 2004, 76, 3179-3184). The formation of more porous silicon Substrate is hydrophobic which makes a primary matrix ions was demonstrated by quantifying Smaller and more concentrated sample spot after solvent the suppression of the matrix signals (MSE score; evaporation. It is possible that this produces homoge McCombie, G. Knochenmuss, R. Anal. Chem. 2004, neous analyte and matrix crystals and efficiention for 76, 4990-4997) and comparing a thin versus a thick mation. An alternate explanation is that the lower num sample. While the MSI score for the thick sample was ber of electrons produced by irradiation of the silicon 0.94 (0.06) which is almost complete matrix suppres results in less charge recombination and therefore sion—the same conditions only gave a MSE score of greater signal. Different target materials and Surface 0.57 (0.08) for a thin sample. The homogeneity of the imaging techniques are being used to explore, ion for sample preparation method was demonstrated for phos mation from Silicon targets.” pholipids samples and crude lipid extracts. It was pos sible to separate groups of mouse serum based on their 0.053 "Matrix-free Laser Desorption Ionization Mass lipid spectra.” Spectrometry using Porous Alumina Thin Film Struc tures.” by Ranu Nayak; and Daniel R Knapp' who stated 0052 “MALDI on Silicon' by Jae-Kuk Kim; and Ker that “Matrix-assisted laser desorption ionization mit K. Murray who stated that Porous silicon substrates (MALDI) mass spectrometry (MS) has been established have been used in the past for Small molecule analyses as one of the key methods for proteomic analysis. with the matrix-free soft ionization method known as MALDI is complicated by the need to have good co desorption/ionization on silicon (DIOS). Non-porous crystallization of sample and matrix. Matrix-free meth silicon targets have been used with infrared lasers for ods of laser desorption ionization (LDI) offer the poten ionization that is similar DIOS in that a matrix is not tial to eliminate this complication. MS from porous required and ionization occurs without interference alumina Surfaces has been reported (R1-Anal. Chem. from low mass background ions. In this study, we dem 77, 5364-69, 2005), however, few details were provided onstrate the benefits of using untreated silicon targets for and the report did not describe a systematic study of the matrix-assisted laser desorption ionization (MALDI). experimental parameters. We report here the LDI MS Samples and matrix are deposited along with matrix on analysis of Some peptides using anodized aluminum a silicon wafer and were irradiated with a UV laser. The (Al) surface on glass Substrates and an investigation of resulting ions are mass separated in a time-of-flight mass the experimental factors controlling the MS signal inten spectrometer. Riboflavin (MW: 376.4 Da, Formula: sity. Al thin films were grown on cleaned Pyrex glass CHNO), bovine serum albumin (BSA, MW: Substrates using a thermal evaporation technique. The as 66.430.3 Da), lyophilized Escherichia coli strain W grown films were Subjected to anodization using a dc (ATCC 9637), and phosphorylase b from rabbit muscle regulated power supply at 50V, 60V, and 90V in a 10 vol. (MW: 97 kDa) were ionized by MALDI on both non % phosphoric acid solution (at room temperature). The porous silicon Substrate and stainless steel Substrate. The resulting porous oxide layer was confirmed by scanning dried droplet method was used to sample preparation for electron microscopy. The anodized samples were coated both substrates. The Bruker Omniflex mass spectrom with gold film by magnetron sputtering. Subsequently, eter can be operated in linear and reflectron mode and different peptides (mass range-500 to 10,000 Da; con uses a 337 nm nitrogen laser (Pulse width: 3 ns, Pulse centration-0.1 to 1.0 mg/ml) were spotted on the con energy: 200 LJ max.) at a repetition rate of 2 Hz for ductive anodized aluminum Surface and air dried. Spots ionization. An accelerating Voltage of 19 kV was were analyzed by a Voyager-DE-STR-MALDI-TOF employed with delayed ion extraction. The typical het mass spectrometer. For comparison, the same peptides US 2015/0309021 A1 Oct. 29, 2015 10

were subjected to conventional MALDI analysis using matched peptides from control spectra to analyze prop C-cyano-4-hydroxycinnamic acid as a matrix. A porous erties of peptides that correlate with peak intensity, structure is necessary for ionization of analytes in including composition and N or C-terminal residues. matrix-free LDI-MS (R1). In the present work we have Since there is a bias in the spectra where smaller peptides grown some hexagonal porous alumina structures using have higher average intensities, we divided the peptide thin films of Al on glass substrates. The as grown Al film masses into four distinct mass ranges for independent thickness were 0.3, 0.5 and 0.7 Lum. Utilizing Al film analysis. In each mass range, we divided peaks into three made the anodization process trivial by not being con intensity categories (high, medium and low) based on cerned on the tedious process of mechanical and elec thresholds picked to create an equal distribution of inten trochemical polishing which is an essential pretreatment sities in each category for each mass range. Comparing step for anodizing directly on bulk Al substrates. In other the amino acid, distribution in real peptides versus ran (non-MS) work on nanostructured alumina films, multi domly-matched peptides revealed a strong negative cor step anodization processes are often used on Al Sub relation between peak intensity and the presence of both strates to obtain more uniform porous structures. How cysteine (C) and tryptophan (W). On the other hand, ever, we have shown some promising results using a valine (V) and methionine (M) tend to be favored in high single step anodization process on the Smooth surfaces intensity peaks. Though we expected to see the acidic of Al film with no pretreatment step. Thickness variation residues aspartic acid (D) and glutamic acid (E) corre and anodization at different Voltages have been exam sponding with high peak intensity due to their negative ined to see the effect of pore depth and diameter on MS charge, no such trend was observed. At the N-terminal signal intensity. Samples anodized at 50V gave no pep position, Some of the same trends held as for composi tide signals. Films anodized at 60V and 90V gave peak tion, though with significant differences. Methionine intensities comparable to those with conventional showed an opposite, negative correlation with peak MALDI, however, higher laser intensity was required. intensity when at the N-terminus. Proline (P), tryp Both 60V and 90V anodized samples showed similar tophan (W), and cysteine (C) were also disfavored, results indicating less effect of pore diameter within a whereas valine (V), threonine (T), histidine (H), pheny range of 75 to 120 nm. Thickness (depth of pores) com lalanine (F), isoleucine (I), and alanine (A) all favored parison Suggested that a 0.3 um film is not suitable for peaks of high intensity when at the N-terminus. Of the MS analysis, as the laser flux tends to burn out the two residues found at the C terminus of tryptic peptides, Surface with each shot and subsequently decreasing the arginine (R) was correlated with high intensity peaks signal intensity. Degradation of signal intensity with whereas lysine (K) was found more commonly in low time was also studied. A 15 days old alumina film did not intensity peaks. Our data indicate that composition of show any change in signal intensity and further investi peptides plays a significant role in peak intensity and gations are ongoing with older samples. These results hence the likelihood of peptide observation in MALDI Suggest that porous alumina on glass can be a potential TOFMS. These data can be used as an additional feature matrix-less approach for LDI-MS proteomic analysis.” to improve protein identification methods.” 0054 An statistical analysis of factors related to ion 0055 “Advances in MALDI of small molecules using suppression in MALDI-TOF mass spectrometry” by surfactants as matrix co-additives', by David C. Grant; Dongmei Yang; Michael S. Wisz: Kevin Ramkissoon; and Robert J. Helleur, who stated that "Matrix-assisted Morgan C. Giddings who stated that “Protein identifi laser desorption/ionization time-of-flight mass spec cation relies heavily on the measurement of peptides by trometry has been innovative in the analysis of macro a mass spectrometer, yet ion Suppression impedes pep molecules. However, it is still developing as a technique tide measurement by causing a differential in their spec for Small molecule analysis. The main reason for this is tral intensities that can result in some peptide peaks the abundance of matrix-related ions in the low mass being undetectable. The differing chemical properties of range of the spectra. The Surfactant cetyltrimethylam peptides are thought responsible for this phenomenon. monium bromide (CTAB) has previously been used by We examined the correlations between observed peptide another group as a matrix Suppressor. When added to the intensities and their amino acid sequence content for matrix C-cyano-4-hydroxycinnamic acid (CHCA) at an MALDI-TOF spectra in order to provide a predictive appropriate molar ratio it has been shown to selectively measure for the likelihood that a given peptide might be Suppress matrix ions. Our research expands on this observed, given its sequence. These statistics may be effect by examining a wider variety of surfactants for the used as an additional measure that can be factored into analysis of peptides, cyclodextrins, and Small phenolic the search engines to increase the accuracy of protein compounds such as flavonoids and flavones. Experi identification, particularly for peptide mass fingerprint ments were performed on a Voyager DE-PRO MALDI (PMF) data. We obtained TOF spectra on an ABI 4700 TOF. Analytes were dissolved in 4:1 MeOH:HO (v/v) mass spectrometer, corresponding to trypsin-digested and pre-mixed with CHCA matrix. Each analyte was spots taken from coumassie stained gels of E. coli cyto tested separately with seven surfactants: hexyltrimethy solic extract. Proteins were identified by PMF analysis lammonium bromide, dodecyltriethylammonium bro using Mascot and GFS searching tools, resulting in 350 mide, cetyltrimethylammonium bromide (CTAB), tet matched proteins with significance p-0.05. We also gen rabutylammonium bromide, decamethonium bromide, erated 30,000 control spectra by picking random sets of Triton X-100 and sodium dodecyl sulfate (SDS). Each peaks (mass, intensity) from the entire set of real spectra, Surfactant was mixed with the analyte-matrix solution at Selecting 350 whose match scores were at a significance varying mole ratios to examine the effect of Surfactant level of p-0.05. We compared peptides and their match concentration of analyte signal. The parameters moni ing sequences from the real spectra to randomly tored including analyte resolution, signal-to-noise, peak US 2015/0309021 A1 Oct. 29, 2015 11

area and intensity. Mixtures of analytes were then tested ration. For every peptide?protein tested between the with each Surfactant. Finally, analysis of phenolic com MW's 1,200 to 150,000 Da a signal enhancement of 1-2 pounds in wine, green tea, and blueberry extracts were orders of magnitude was achieved when using selected undertaken to illustrate real sample applications. Mass Sublayer compounds. The most significant enhance spectra of standard peptides, phenolic acids and other ments were observed for the higher MW proteins. Fur low mass molecules (m/z<500) demonstrate improved thermore, consistent with predictions of the equilibrium mass resolution by 15-45% using appropriate Surfac model of MALDI, the preliminary experiments also tants. Surfactant-mediated MALDI-TOF-MS has also showed a strong dependence of the Sublayer enhance been evaluated for Small molecule quantitative analysis ment on the IE of the sublayer being greater than that of using a second analyte as an internal standard. In one the MALDI matrix. This effect is consistent with the experiment, p-coumaric acid (m/z, 147.30) was analyzed possibility of the Sublayer acting as a "charge sink’ and using quercetin (m/Z303.45) as an internal standard and inhibiting protonation of the analyte i.e. if the IE of the the resulting calibration curve displayed good linearity sublayer is lower than that of the MALDI matrix then an (R=0.9872). Surfactants tested showed promising electron transfer between the photoionized matrix mol results as matrix-ion Suppressors, as spectra were mostly ecules and the sublayer will occur leading to loss of the free of all matrix ions, and displayed low background matrix ions necessary for analyte protonation. Finally, noise at the appropriate molar ratio. It was found that a preliminary experiments suggest that SA-MALDI will matrix:analyte:surfactant ratio of approximately 1000: be particulary useful for complex mixture analysis. 20:1 gave the best spectra in most cases. In some cases, Enhancements of both low MW and high MW signals of ion Suppression was observed by using even lower up to an order of magnitude are observed for both control amounts of surfactant (10000:20:1). Surfactant-medi mixtures of peptides/proteins and for the complex pro ated MALDI showed promise in analysis of analyte tein mixture derived from the cyanobacteria source.” mixtures as well as real samples containing phenolic 0.057 “Sensitive and reproducible detection of peptides compounds. Several were identified and Some quantita by Maldi-TOF MS using self-assembled monolayer tively measured by MALDI. Their identity was con (SAM)-based chips—specifications and applications.” firmed by LC-MS. These results indicate that surfactant by Udo Roth; Holger Röhl; Matthias Gluckmann; mediated MALDI may be a good technique for analysis Thorsten Jaskolla; Michael Karas; Kerstin Steinert; and of small molecules.” Karsten Reihs, who stated that “Sensitivity, mass accu 0056 “Sublayer Assisted MALDI (SA-MALDI): Prac racy, and reproducibility are prerequisites for efficient tical Applications and Fundamental Studies.” by Zaneer MALDI-MS analyses used in current quantitative pro M. Segu; and Gary R. Kinsel, who stated that “MALDI teomic work-flows. Our approach consists of a novel analyses of complex proteomic mixtures Suggest that technology for peptide sample preparation using plates desorption/ionization of high mass proteins is much less with pre-deposited matrix spots of extra-fine Sub-micron efficient than for lower mass peptides. The equilibrium size CHCA crystals prepared by vacuum sublimation on model of the MALDI process Suggests that analyte ion an ultraphobic surface. Such a design enables the reli ization proceeds via thermodynamically driven gas able application of nanoliter Volumes of analyte solu phase ion-molecule reactions within the plume of laser tions and facilitates highly reproducible signal detection desorbed material and, hence, that the primary limitation in the low attomole range. To provide a platform for to obtaining MALDI ion signals from high MW proteins high-resolution LC-MALDI-TOF, a high-density array is likely related to inefficient desorption of these com chip accommodating up to 1600 spots is Supplied. We pounds. If true, alternate approaches that lead to present the results of our investigation on the effect of improved desorption of these species within the MALDI the properties of the CHCA matrix crystals on the initial plume should yield enhanced high molecular weight Velocity of the peptide ions and their fragmentation in protein MALDI ion signals. We have explored this MS/MS experiments. Sensitivity, mass accuracy, and hypothesis through the use of various desorbable sub reproducibility were assessed using different volumes layers onto which conventional protein MALDI samples and concentrations of a peptide mixture spotted onto a are deposited. Sublayers for SA-MALDI were chosen Mass SpecTurbo Chip with 192 spots. Special attention from a group of hydrophobic polyaromatic hydrocar was paid to the flatness of the target and its proper bons. The sublayers were chosena) to be insoluble in the electrical ground connection. For LC-MALDI analysis, matrix and analyte solvents, b) to absorb at 337 nm, c) to tryptic digests were separated using a reversed phase have no easily donatable protons and d) to have a range (C18) HPLC-column. Samples were spotted (Probot, of IEs above and below those of typical MALDI matri LC Packings) on a high-density Mass SpecTurbo Chip ces. MALDI MS and SA-MALDI MS analyses were (1600 spots). Prior to MS analysis, the air-dried spots performed using various peptides and proteins ranging were treated with 15 mMammonium dihydrogen phos in MW from 1,200 to 150,000 Da. Conventional dried phate to efficiently reduce the formation of matrix clus droplet sample preparation was employed, either ter ions. The initial velocity was determined by the directly on the stainless steel MALDI target or on the delayed extraction method in a linear two-stage accel sublayer coated MALDI target. Similar experiments eration TOF system. Experiments were carried out using were performed using peptide?protein mixtures and, MALDI-TOF instruments from Bruker (Reflex III) and finally, using a crude protein extract from cyanobacteria. Applied Biosystems (DE-Pro, 4700, 4800). In the sen The preliminary studies performed clearly demonstrate sitivity trial, detection of peptide amounts, which were that a significant enhancement in the peptide?protein applied from a standard dilution series, was achieved MALDI ion signal is achieved for SA-MALDI as com down to the attomole? spot range. A mass accuracy of an pared with the conventional dried droplet sample prepa average mass deviation of less than 10 ppm for each spot US 2015/0309021 A1 Oct. 29, 2015 12

was routinely obtained by external mass calibration were monitored to avoid matrix signal interference. using the four corner spots of the array. MS/MS experi Results from the OLZ (m/z. 313->256) dosed tissue ments analyzing attomole amounts of peptides delivered showed that signal was seen with both CHCA and DHB fragment spectra that provided sufficient information for matrices; however, optimal sensitivity by an order of the unambiguous identification of peptides. Reproduc magnitude was achieved using CHCA in 50% acetoni ibility of sample preparation yielded RSD values of trile. The model predicted the same optimal matrix/sol typically less than 10-20% depending on the peptide. vent as for OLZ dosed tissue, but demonstrated a two The benefits of sensitivity, mass accuracy, and reproduc fold increase in sensitivity not seen in the dosed tissue. ibility provide an optimal combination for high through Similar drugs within the benzoazepine class. Such as put LC-MALDI experiments. We show that peptide carbamazepine (m/Z 237-> 194) and clozapine (m/z identification can be significantly increased compared to 328->272), preferred CHCA. Drugs in the f-andrener standard procedures using stainless steel Substrates. Due gic blocker class, such as PPL (m/z. 260-> 116) and to the Sublimation deposition technique used for matrix metaprolol (m/z. 268-> 116), exhibited preference for spot preparation, we obtain very small CHCA crystals of CHCA based on the model; however, no clear difference a typical crystal size of less than 1 micron. We found that in sensitivity was observed for various solvent condi these spots yield significantly higher initial Velocities of tions. This was also seen in the PPL dosed tissue. Dosed the emitted peptide ions in the range of 650 m/s com SPN tissue (m/z 396->165), indicated an affinity for pared to about 300 m/s for standard dried droplet prepa DHB independent of solvent. This preference was also rations. Higher initial velocities may significantly influ predicted by the model. Preliminary data indicates that ence fragmentation processes by more effective matrix/Solvent trends do exist within a drug class. The expansional cooling during ion formation. Results con advantage of the model allows us to effectively mimic cerning the relationship between the initial velocity and the behavior of dosed tissue, while avoiding the need to the peptide fragmentation process will be presented and dose and sacrifice animals for each determination of discussed.” optimal matrix/Solvent combination. The model is being 0.058 “Predicting matrix conditions for drug analysis in extended to butyrophenone (SPN) and B-andrenergic tissue by MALDIMS/MS” by Sara L. Frappier. Sheerin blocker (PPL) classes in addition to other drug classes, K. Shahidi; and Richard M. Caprioli' who stated that “It as well as, other tissues such as kidney and brain.” has been previously found that various drug classes have 0059 Focusing chips for MALDI-MS have different preferential matrix/solvent conditions when analyzing requirements than focusing chips for X-ray fluorescence. dosed tissue by MALDI MS/MS. Since there are no Focusing chips for X-ray fluorescence should preferably not established trends known for matrix/solvent preference interfere with XRF measurement. For example, the chips for Such analyses, time consuming optimization proce should have a limited amount of chemical elements that may dures are needed before a study can commence. The goal interfere with the sample being measured. Preferably the of this study was to determine the trends of matrix/ focusing chip for use with X-ray fluorescence should have an Solvent preference for drugs within a class. A model has amount of a chemical element that produces less than 25% of been developed that provides a method for analyzing a the signal of the sample being measured, when measured for large number of drugs within a class and across classes. at least one of the elements that are present in both the focus Using the identified trends, sample preparation of dosed ing chip and the sample. More preferably, the focusing chip tissue will have a guideline of matrix/solvent preference for X-ray fluorescence is free of elements that may interfere for analysis by MALDI MS/MS, saving time and with the sample being measured. Typical elements to be mea resources. Liver tissue was excised from control (non Sured include Sulfur, fluorine, phosphorus, chlorine, bromine, dosed) and Olanzapine (OLZ; 8 mg/kg), Spiperone iron, Zinc, magnesium, calcium, titanium, Scandium, and (SPN; 5 mg/kg) or Propranolol (PPL: 10 mg/kg) dosed platinum. The metal Substrate that the focusing chip is Sup Fischer 344 mice. Using a cryostat, tissue was sectioned ported on should not contain elements that have X-ray emis (20 um) and thaw-mounted onto gold-coated MALDI sion spectral peaks in the region of interest, and preferably do target plates. For the prediction model, 1 Jul of 1.5 g/ml not have X-ray emission spectral peaks between 2 KeV and 3 of a drug standard was spotted on to a MALDI plate, KeV. allowed to dry, and then covered with control liver tis sue. Sinapinic Acid (SA), 2,5-dihydroxybenzoic acid 0060 Focusing chips for X-ray fluorescence should pref (DHB) and C-cyano-4-hydroxycinnamic acid (CHCA) erably be thin. Thick chips or slides cause scattering in the matrices were prepared at 25 mg/ml into various binary X-ray portion of the electromagnetic spectrum. Preferably, combinations of acetonitrile, methanol, isopropanol and the chips are less than 500 microns thick, and more preferably water. Matrix combinations were spotted onto control the chips are less than about 50 microns thick. Focusing chips and dosed tissue sections for MS/MS analysis by an as described in the MALDI-MS literature, for example from oMALDI-Qq-TOF-MS. Purportedly, matrix/solvent the list of presentations at the 54th ASMS Conference on preferences rely on extraction of drug from tissue and Mass Spectrometry, May 28-Jun. 1, 2006, Washington State the drugs ability to co-crystallize with matrix. Our Convention & Trade Center, Seattle, Wash., should be re model mimics extraction of drug from dosed tissue. In manufactured on thin supports to facilitate their use with order for drug signal to be detected, the Solvent must X-ray fluorescence. extract the drug through the tissue so that it may co 0061 The size of each hydrophilic region is important. crystallize with the matrix. Dosed tissue sections were Preferably, each hydrophilic region has an area between 75 spotted with each matrix/solvent combination to deter square picometers (equivalent to a spot with a diameter of mine the optimal conditions for analysis. For each drug about 10 microns) and 3.6 square microns (equivalent to a analyzed, transitions from parent to major fragmentions spot with a diameter of about 4 millimeters). US 2015/0309021 A1 Oct. 29, 2015

0062 One convenient method for preparing focusing ment with a monocapillary focusing optic, polycapillary chips uses polymer assisted deposition (PAD), described in: focusing optic, a collimator, a microfocus X-ray tube, a syn Published Patent Application 2005.0043.184, “Polymer-as chrotron X-ray source, a linear accelerator X-ray source, a sisted deposition of films.” published Feb. 24, 2005, to Tho rhodium X-ray tube, a molybdenum X-ray tube, a chromium mas M. McCleskey et al.; Published Patent Application X-ray tube, a silver X-ray tube, a palladium X-ray tube, a 20050008777, “Polymer-assisted deposition of films,” pub monochromatic X-ray source, a polychromatic X-ray source, lished Jan. 13, 2005, to Thomas M. McCleskey et al.: “Poly a polarized X-ray Source, a confocal X-ray fluorescence spec mer-assisted deposition of metal-oxide films.” Jia, Q. X. trometer, a PIN diode detector, a semiconductor X-ray detec McCleskey, T. M.; Burrell, A. K. Lin, Y.; Collis, G. E.; Wang, tor, a germanium or doped germanium X-ray detector, a sili H. Li, A. D. Q.: Foltyn, S. R. Nature Materials 2004, 3(8), con or doped silicon X-ray detector, a wavelength dispersive 529-532.: “Epitaxial growth of Eu2O3 thin films on LaAlO3 X-ray fluorescence spectrometer, an energy dispersive X-ray substrates by polymer-assisted deposition Lin, Y.; Wang, H.; fluorescence spectrometer, total reflectance X-ray fluores Hawley, H. E.; Foltyn, S. R. Jia, Q. X. Collis, G. E.; Burrell, cence spectrometer, and the like. A. K. McCleskey, T. M. Appl. Phys. Lett., 2004, 85(16), 0067. If total reflectance x-ray fluorescence (TXRF) is 3426-3428; "Structural and dielectric properties of epitaxial employed with the present invention, the preferred substrates Ba1-xSrxTiO3 films grown on LaAlO3 substrates by poly are quartz, lucite, mylar, silica, Silicon with a smooth Surface mer-assisted deposition Lin, Y.; Lee, J-H.; Wang, H. Li, Y.: for sample deposition. Foltyn, S.R., Jia, Q.X.; Collis, G. E.; Burrell, A. K.; McCles key, T. M.: Appl. Phys. Lett., 2004, 85(21), 5007-5009: Beam Size “Green luminescent zinc oxide films prepared by polymer assisted deposition with rapid thermal process” Lin, Y.; Xie, 0068. In X-ray fluorescence microscopy, we have found J.; Wang, H. Li, Y.: Lee, S.Y.; Foltyn, S. R.: Crooker, S.A.: that the most efficient (strongest signal with lowest noise) Jia, Q. X.; Burrell, A. K. McCleskey, T. M. Thin Solid Films, X-ray beam size, ceteris paribus, is one that is essentially 2005, 492(1-2), 101-104.: "Conformal coating of nanoscale matched to the sample size. This situation has not arisen features of microporous AnodiscTM membranes with zirco previously because the samples used in the past have been nium and titanium oxides.” Shukla, P.; Minogue, E. M.: larger than the beam size, so many systems use a small beam McCleskey, T. M.; Jia, Q. X. Lin, Y.: Lu, P.; Burrell, A. K.: size and rastering or scanning across the sample. Also, many Chemical Communications, 2006, (8), 847-849.: “Polymer samples are not homogenous, so having a beam size matched assisted deposition (PAD) of thin metal films: a new tech to the sample size has not previously made much sense. nique to the preparation of metal oxides and reduced metal 0069. For the biomedical samples we have analyzed, the films.” Shukla, P.; Lin, Y.; Minogue, E. M.; Burrell, A. K.: samples are generally circular and tend to have diameters McCleskey, T. M.; Jia, Q.: Lu, Pi. Materials Research Society between 40 um and 2 mm (i.e., areas of about between 1 Symposium Proceedings 2006, 893, 183-188; all incorpo square nanometer and 3 square microns). These samples are rated herein by reference. The PAD process may be used to also homogenous or otherwise have no need to be sampled in deposit patterns of different materials, including metallic Subregions. In Such cases, we have found that matching the phase metals, metal oxides, and metal nitrides. Using PAD, beam size to the spot size provides the strongest signal to different regions of hydrophilic materials and hydrophobic noise ratio. Thus, a preferred X-ray fluorescence source for regions on a Substrate, typically a silicon or aluminum Sub measuring a sample comprises an X-ray Source, where the Strate. transmitted X-ray beam has a cross-sectional area essentially 0063 Another convenient method for preparing focusing the same surface area as the sample. Thus the beam preferably chips uses the formation of hydrophilic and hydrophilic has a cross-sectional area of between about 1 square nanom regions on a Substrate using Supported mesoporous silica. eter and 3 square microns. “Matching” would preferentially This method is described in A. M. Dattelbaum and S. lyer, be as close as match as practical, but an imperfect match "Surface-assisted laser desorption ionization mass spectrom where the beam has a cross-sectional area of between 25% etry.” Expt. Rev. Proteomics. 2006, 3(1): 153-61; and in U.S. and 250% of the surface area of the sample is also acceptable. Pat. No. 6,958,480, “Sample desorption/ionization from 0070 This area matching may be accomplished in rare mesoporous silica”, published Oct. 25, 2005 to Srinivas lyer cases by having an X-ray source whose transmitted beam and Andrew Dattelbaum; and Nancy H. Finkeletal, “Ordered cross-sectional area matches the sample area without any Silicon Nanocavity Arrays in Surface-Assisted Desorption/ modification of the beam. More typically, the beam size will Ionization Mass Spectrometry” Analytical Chemistry, 77 (4), have to be modified to match the sample size. Modification is 1088–1095, 2005 incorporated herein by reference. typically carried out by means of focusing optics or by a 0064. In each of the cases described above, the MALDI collimator. Thus, the source further comprises a focusing focusing chips is preferably modified such that it is formed on means or collimator disposed between the source and the a thin substrate, preferably less than 500 microns thick and sample. more preferably less than 50 microns thick. 0065. The XRF focusing chip is used by depositing a Sample Composition sample, which typically consists of an aqueous sample of a 0071. Many samples, such as proteins, require a buffer to protein or nucleic acid, which has been optionally modified maintain the pH within a particular range (i.e. a pH buffer), or by addition of an inhibitor, co-factor, metal, protein, Sugar, or to maintain the redox state of a chemical (i.e. a redox buffer). other chemical. The sample is allowed to dry, and then the Many buffers contain elements which might interfere with the focusing chip with the deposited Sample is irradiated with measurement of the chemical. The buffer should preferably X-rays and any fluorescence of X-rays are measured. be free of at least one chemical element having an atomic 0066. The sample on the X-ray focusing chip may be number of greater than four, where that chemical element is conveniently measured using an X-ray fluorescence instru present in the chemical. The buffer should also preferably be US 2015/0309021 A1 Oct. 29, 2015

free of at least one chemical element having an atomic num desalting well plates and DNA detaining well plates. Separa ber of greater than four, where that chemical element is tions are preferably carried out in less than 30 seconds at 4 present in the receptor. The buffer should more preferably be degrees Celsius, as described in Annis DA et al. An affinity free of at least one chemical element having an atomic num selection-mass spectrometry method for the identification of ber of greater than eight, where that chemical element is Small molecule ligands from self-encoded combinatorial present in the chemical. The buffer should also more prefer libraries: Discovery of a novel antagonist of E. coli dihydro ably be free of at least one chemical element having anatomic folate reductase. International Journal of Mass Spectrometry. number of greater than eight, where that chemical element is 2004, 238:77-83. An alternative method of separation is ultra present in the receptor. The buffer should be free of surfac filtration, Such as is described in the following examples: tants or detergents that contain Sulfur or phosphorus. The buffer should preferably be free of at least one of the follow First Example ing chemicals or functional groups: dimethylsulfoxide, thi ols, sulfate anion, Sulfonate anions, chloride anion, bromide 0074. We measured the interaction between biotin and anion, fluoride anion, iodide anion, perchlorate anion, phos avidin to demonstrate that RPM could quantify the sulfur phate anion, and phosphonate anions. The buffer preferably containing small molecule (biotin, CHNOS) over the does contain one or more of the following chemical or func Sulfur content of a protein (tetrameric avidin, Sigma-Aldrich, tional groups: amine, imine, nitrate anion, nitrite anion, 16 sulfur atoms, see Korpela J. Avidin, a high affinity biotin ammonium cation, and iminium cation; these chemicals offer binding protein, as a tool and Subject of biological research. the correct buffering properties with minimal x-ray fluores Medical Biol. 1984, 62:5-26). Previous studies have shown cence interference. Most preferably, the buffer consists of an that the ratio of sulfur in avidin-biotin:avidin is 19.5:16, or ammonium nitrate Salt Such as tris(ethanol)amine nitrate, also 1.22, see Green N. Mex. Purification of Avidin. Meth Enzy known as tris nitrate. mol. 1970, XVIII(A):414-417. We incubated 500 micro 0072 The proteins to be used with the present invention grams of avidin in 1.0 ml of 0.20 millimolar biotin at pH 8.0 are preferably purified to remove chemicals which are not for 5 minutes and then centrifuged the sample for 3 hours at chemically bound or which are loosely bound to the protein. 7000 g using a Centricon YM-3 resulting in a concentrated Loosely bound is defined in this paragraph as meaning having avidin-biotin sample of 25-50 ul. We diluted the sample to a binding affinity that is weaker than about one (1) millimolar. 0.75 ml in TRIS buffer (pH 8.0), centrifuged for 2 hours at This desalting step may be conducted conveniently using gel 7000 g, and repeated this process twice. We treated an avidin filtration chromatography or size exclusion chromatography, control sample identically. We used RPM to measure the such as using a Sephadex G25 or Sephadex G50 column. sulfur content of the avidin-biotin complex and the avidin. We Desalting may also be accomplished using dialysis, including confirmed that the amount of avidin in each sample was Vibratory Shear Enhanced Process dialysis described in identical using bicinchoninic acid protein assay kit (Pierce). http://www.vsep.com/pdf/PolymerDiafiltration.pdf. This As predicted, the avidin-biotin complex contained 1.2 times process is amenable to multiplexing using a well plate format, the sulfur than the avidin alone as measured by RPM. This set such as a 96-well, 384-well, or 1536-well plate format. Sepa of experiments demonstrated that RPM can measure strong rations systems such as Zeba 96-well plates available from non-covalent interactions and use fluorescence ratios to deter Pierce Biotechnology Inc., PO Box 117, Rockford, Ill., mine binding even when m the protein and analyte have only 61105, are particularly convenient. As can be seen in the a shared RPM atom (in this case, sulfur). following table, Zeba well-plates function across a wide range of concentrations of Bovine Serum Albumin, where the Second Example units for the numbers in the table are percent mass recovered, 0075. Our preliminary data show that RPM measures non unless otherwise noted: covalent protein-inhibitor complexes. We used di-Zinc ami nopeptidase (aAP) and its competitive inhibitor leucine phos phonic acid (LPA). LPA binds to aAP with a Ki of 6.6 uM at 10 25 micro- 50 pH 8.0, see Stamper Cetal. Inhibition of the aminopeptidase 5 micromolar micromolar molar micromolar from Aeromonas proteolytica by L-leucinephosphonic acid. 10 microliters 39 +f- 21% 48 +f- 2%. 63 +f-5%. 62 +f-3% Spectroscopic and crystallographic characterization of the 20 microliters 79 -f- 11% 70+f- 2%. 78 +f- 2%. 83 +f- 2% 30 microliters 87+f-3% 79 +f- 4% 80+f- 2% 90+f- 4% transition state of peptide hydrolysis. Biochemistry, 2001, 40 microliters 103 +f-3% 95+f- 1% 89+f- 8% 98+f-5% 40.7035-7046. We incubated 500 micrograms of aAP in 1.0 mls of 0.20 millimolar LPA (>10xKi) at pH 8.0 for 5 minutes. We centrifuged the aAP-LPA sample for 3 hours at 7000 g 0073. The single-plexed or multiplexed gel filtration chro using a Centricon YM-3 resulting in a concentrated sample of matography process may be expedited using a centrifuge, 25-50 ul. We diluted this sample to 0.75 ml in TRIS buffer (pH such as a the IEC Multi and Multi RF Centrifuges with a 8.0), centrifuged for 2 hours at 7000 g, and repeated this Microplate Rotor Available through Fisher Scientific, catalog process twice. We Subjected two negative control samples to #05-375-77, Liberty Lane, Hampton, N.H. 03842, manufac the same filtering conditions described above: The first con tured by International Equipment Company; or a vacuum trol was 1.0 ml of 0.20 millimolar LPA, and the second manifold. Such as a Vacuum apparatus Such as the Multi control was 200 micrograms of avidin and 1.0 ml of 0.20 Screen Vacuum manifold with Direct Stack from Millipore, millimolar LPA to test the possibility of electrostatic and 290 Concord Road, Billerica, Mass. 01821, attached to a hydrophobic interactions between LPA and non-specific pro standard vacuum pump (Catalog # WP61 11560) also avail tein. We measured phosphorus was measured in the aAP-LPA able from Millipore. Separation may be performed using complex in a 1:2 ratio; no phosphorus was observed in the ZipPlate(R) micro-SPE Plate and MALDIspot TM Accessories control samples. This set of experiments demonstrated that also available from Millipore, as well as similar protein RPM can measure weak non-covalent interactions (6.6 US 2015/0309021 A1 Oct. 29, 2015

micromolar) and use fluorescence ratios to determine binding DDT2005-2.ppt. While no ideal solution has been found, when the protein and analyte have different RPM atoms (in preferable solutions include the following. One preferable this case, Zinc and phosphorus). approach to protein deposition uses focusing chips (described above), which promote even drying of protein Solutions. Sample Deposition Another convenient approach to protein deposition is to 0076 Samples may be conveniently deposited on sub digest the protein using pepsin or trypsin or other proteases; strates in several ways. One very convenient manner for this approach changes the Surfactant properties of the protein sample deposition is deposition of a Volume of Solution con and promotes more even drying of samples. Other matrix taining the sample using a pipette or Syringe. Pipettes are modifying chemicals, such as acetic acid, trifluoracetic acid, especially preferred because they may be multiplexed. Such detergents, salts (especially salts that do not contain some or as an EppendorfR Research Multichannel Pipette catalog all of the chemical elements being measured in the chemical number 0224.52002, available from Eppendorf North or the protein) or Surfactants, promote the even drying of America Inc. One Cantiague Road, P.O. Box 1019, Westbury, samples; these matrix modifiers are thought to cause proteins N.Y. 11590-0207. The amount of sample to be deposited is to denature or to adopt a molten globular state by disrupting important. Large sample solution Volumes tend to dry slowly protein folding through the disruption of hydrogen bonds, salt and unevenly. Small sample Volumes contain only small bridges, disulfide bonds, and other covalent and non-covalent amounts of sample. Preferably, the sample solution is depos interactions that affect protein folding. This molten globular ited in a volume of between 1 nanoliter and 50 microliters, state must be achieved without precipitation of the protein, and more preferably the sample solution is deposited in Vol which leads to inhomogeneity in the sample. Proteins may umes of between 10 nanoliters and 10 microliters. Most pref also be denatured or rendered into their molten globular state erably, the sample solution is deposited in Volumes of by heating the solutions of the sample, preferably above 90 between about 25 nanoliters and about 8 microliters. The degrees Celsius, but at least above about 40 degrees Celsius, sample concentration is also important. The sample solution depending on the protein and its thermal stability. High qual that is deposited preferably contains between 100 picograms ity protein spots may be generated by heating under an infra of protein and 1 gram of protein. More preferably, the sample red lamp, or a Sun lamp, or a halogen lamp, or any lamp with Solution that is deposited contains between about 1 nanogram a wattage above about 55 watts. High quality protein spots of protein and 1 milligram of protein. The reason for this may also be obtained when the sample is dried while being sample mass requirement is to balance the cost of protein and centrifuged, preferably also under reduced pressure. Cen the measurement requirements of the technique. The mea trifugation under at least a partial vacuum is particularly Surement technique can measure samples containing as little convenient for samples that are multiplexed, because the as about 100 femtograms of the chemical elements to be whole set of multiplexed samples may be dried under identi measured. More conveniently, measurements are obtained on cal conditions. samples containing between ten nanograms and one micro gram of the element to be measured. Robotic pipettes or X-Ray Sources automated biological printers may be used, such as a BioDot BJQ3000 BioJet Quanti 3000 or BJP3000 BioJet Plus 3000, 0078. Many X-ray sources will function with the present available from BioDot, Inc, 17781 Sky Park Circle, Irvine, invention. Rhodium and molybdenum X-ray Sources are com Calif. 92.614. Acoustic printers may be conveniently used, monly employed. Biological samples and pharmaceutical such as the Echo 555 Liquid Handler, the Echo 550 Liquid samples frequently contain the elements phosphorus, Sulfur, Handler, and the PortraitTM 630 MALDI Reagent Multi-Spot and chlorine. It is preferable that the X-ray sources do not ter, available from Labcyte Inc., 1190 Borregas Avenue, generate noise or excessive scattered X-rays in the region of Sunnyvale, Calif. 94089. the spectrum that is of most importance. Therefore, it is 0077 Samples should be dried in a manner that promotes preferable to use a chromium, palladium or silver X-ray formation of an evenly distributed spot. Drying a sample of source. If the sample is known or believed not to contain protein into an evenly distributed spot is extremely challeng chlorine, or if the chlorine spectrum is not needed for the ing. Proteins affect the Surface tension of water, especially at analysis, then a rhodium source may be used. Molybdenum high protein concentrations and Small solvent Volumes, i.e. X-ray sources are less efficient than other X-ray tubes at excit the exact conditions for drying. This problem is well known in ing phosphorus, Sulfur, and chlorine, and are less preferred. the art, and has prompted the development of highly expen Most preferably, the X-ray tube has a characteristic K-alpha or sive solutions such as the Labcyte acoustic equipment men L-alpha line at or above about 2.838 KeV and less than about tioned above, and described in “The Echo 550tm Delivering 9.441 KeV. X-ray M lines are frequently broad and less effi Results in Screening.” by Jennifer Zewinski, available at cient. The X-ray source preferably does not have an M line http://www.labcyte.com/aboutus/publications/Zewinski above 2.120 KeV. X-rays that impinge on the sample may be (BMS)EchoPresentation.pdf: generated by X-ray tubes directly, or indirectly by the excita and in Acoustic Non-Contact Dispensing The Right tion of a target by X-rays. Choice for UHTS. By Timothy P. Spicer, available at http:// www.labcyte.com/news/events/Spicer %20Presentation Chemicals %20for%20SBS 96202004%20revised%208 19 041 ppt: and by “Pharmacological Evaluation of Different Compound 0079 While this process may be used for many chemicals, Dilution and Transfer Paradigms on an Enzyme Assay in Low it is preferable that the chemical to be measured comprises at Volume 384-well Format.” by Tim Spicer, Yvonne Fitzgerald, least one of the following chemical elements chemically Neil Burford, Sandra Matson, Moneesh Chatterjee, Mark bound to the majority of the chemical: fluorine, phosphorus, Gilchrist, Jim Myslik and Jonathan O'Connell, available at Sulfur, chlorine, bromine, iodine, platinum, copper, iron, Zinc, http://www.labcyte.com/aboutus/publications/BMSposter gallium, or gadolinium; in this context, chemically bound is US 2015/0309021 A1 Oct. 29, 2015

defined as covalent bonds or metal-ligand bonds having a mide; Tacrolimus; Salmeterol: Clofazimine: Alprazolam: bond strength of at least 30 kCal/mole. Moxifloxacin; Vigabatrin; Mitomycin; Cefuroxime; Tridi 0080 Chemicals which may be used with the present hexethyl; Tropicamide; Amiodarone; MometaSone; Thiogua invention include the following, along with any salts and free nine; Lomustine; Gemfibrozil; Bumetanide; Torsemide; bases: Chlorpheniramine; Methimazole; Nelfinavir, Zonisa Famotidine: Propylthiouracil; Isradipine: Flucytosine: mide; Naltrexone; Carmustine: Ifosfamide: Rizatriptan; Mefloquine; Nadolol; Ropinirole; Pentamidine: Tirofiban: Ramipril; Milrinone; Tenoxicam; Apraclonidine; Neomycin; Sertaconazole; Triprolidine: Clobazam; ChlorZoxazone: Gabapentin; Anastrozole; Alitretinoin. Oxytetracycline: Pra Levodopa; Olopatadine; Estradiol; Ritonavir; Triazolam: Zosin; Amifostine; Desipramine; Hydroxyurea; Naloxone; Methscopolamine; Trimethadione: Remoxipride: Quina Kanamycin; CandoXatril; Tramadol; Chlorpropamide; crine; Ethosuximide: Fenfluramine; Ampicillin; Rivastig Oxaprozin; Trichlormethiazide; Sulpiride: Cyproheptadine: mine; Thiethylperazine; Desloratadine; Piperacillin; Vitamin Brimonidine; Tamsulosin; Misoprostol; Pentolinium; Done B12: Fluconazole; Pravastatin; Aminophylline; Dacarbazine: pezil; Itraconazole; Penciclovir; Bicalutamide: Epinastine; Cinnarizine: Vidarabine; Verapamil: Cromolyn; Carbam Trimethaphan; R-mephobarbital; Clavulanate; Nitrofura azepine; Propiomazine; Prednisone; Warfarin; Methylpred Zone; Bethanechol; Losartan; Gemifloxacin; Clonazepam, nisolone; Clomocycline; Tiagabine; Dapsone; Fluvastatin: Atorvastatin: Heparin; Methohexital; Efavirenz: Duloxetine; Fentanyl; Fexofenadine: Palonosetron; Methotrexate: Cyclizine; Methoxamine; Benazepril; Amsacrine: Flutica Meclizine; Zopiclone: Promazine: Cisplatin: Dipyridamole; sone Propionate; Divalproex; Etodolac. Enoxaparin; Indi Epirubicin; Tretinoin, Esomeprazole; Paroxetine; Phenyle navir; Trihexyphenidyl; Tadalafil; Progabide; Pantoprazole; phrine; Diphenoxylate; Dofetilide; Acrosoxacin: Lovastatin: Meperidine; Guanfacine; Sulfamethoxazole; Lansoprazole; Mitoxantrone; Ibuprofen; Celecoxib; Felodipine; Zolmitrip Porfimer; Triamterene; Cocaine; Ribavirin; Theophylline: tan; Zafirlukast; Zanamivir, Sumatriptan; Pyridostigmine; Vitamin C: Dopamine; Minoxidil; Nicardipine; Phenace Glimepiride; Pilocarpine: Procyclidine: Loratadine; Dutast mide: Dexrazoxane: Carvedilol; Hydrochlorothiazide; Phen eride; Mequitazine: Oxycodone: Flupenthixol; Toremifene: termine; Rifabutin; Zolpidem; Tegaserod; Orphenadrine; Vindesine; Trospium; Pemirolast; Ceftriaxone; Conjugated Digoxin; Phenelzine; Aprepitant; Vinorelbine; Cerivastatin: Estrogens: Azithromycin; Doxepin: Oxyphencyclimine; Ral Trimethoprim; Simvastatin: Argatroban; Norgestrel; Perhex oxifene; Ketoconazole; Nefazodone; Rosiglitazone; Chloro iline; Pefloxacin; Indomethacin; Levobupivacaine; Rescin procaine; Fenofibrate; Physostigmine; Cyclophosphamide; namine; Dicyclomine; Bexarotene; Chlorambucil; Phenylbutazone: Risedronate; Zaleplon; Streptomycin; Irbe Lorazepam; Hesperetin: Melphalan: Acetazolamide: sartan; Entacapone; Carisoprodol: Domperidone: Halofan Codeine; Pentoxifylline; Dobutamine; Tamoxifen; Dactino trine; Candesartan; Nitrendipine; Triamcinolone; Penicillin mycin; Venlafaxine; Idarubicin; Chlorthalidone; Tizanidine: V: Ciprofloxacin; Fluvoxamine; Vitamin D2 (Ergocalcif Flecainide; Uracil Mustard; Dichlorphenamide: Adenosine: erol); Oxybutynin; Calcidiol; Perphenazine: Raltitrexed: Valsartan; Nandrolone Phenpropionate; Ouabain; Quinidine: Eszopiclone: Mifepristone; Testosterone; Montelukast; Methacycline; Olanzapine; Isotretinoin; Balsalazide. Amox Allopurinol; Glipizide. Sulfanilamide: Repaglinide; Stavu apine; Vitamin D4 (Dihydrotachysterol); Exemestane: Rilu dine: Protriptyline; Budesonide; Omapatrilat; Clopidogrel; Zole; Tolterodine: Citalopram; Cidofovir; Delavirdine: Tolcapone: Omeprazole; Zidovudine: Epinephrine; Sulfac Nilutamide: Rofecoxib; Sulfasalazine; Nitroglycerin; Thia etamide: Bleomycin; Cisapride: Isosorbide Dinitrate; mylal; Cilostazol; Pramipexole; Methoxsalen: Oxymor Sibutramine; Phenylpropanolamine: Mecamylamine; Gli phone. Succinylcholine; Carbidopa; Mupirocin; Remikiren: clazide; Cefprozil; Acetophenazine; Methysergide; Phytona Captopril; Levobunolol; Phenindione: Probenecid; Solifena dione; Triflupromazine; Carboplatin: Chloroquine; Nor cin Succinate; Almotriptan; Tolazoline; Arsenic Trioxide; floxacin; Clozapine; Reserpine; Diltiazem; Doxazosin; Atenolol; Trifluoperazine; Clonidine: Sertraline; Tub Brinzolamide: Dihydroergotamine; Levofloxacin; ocurarine; Propantheline; Sirolimus; Prilocaine; Clarithro Lidocaine; Amphetamine; Ondansetron; Chlorpromazine; mycin; Isoproterenol; Valdecoxib; Phenobarbitone; Doxoru Telithromycin; Methadone; VitaminA, Guanabenz; Troglita bicin: Oxaliplatin: Risperidone: Proguanil; Oxyphenonium; Zone; AlfuZosin; Sulfapyridine; Ropivacaine; Ketamine; Sulfadiazine; Nitrofurantoin: Lercanidipine: Propranolol; Mitotane; Vincristine; Phensuximide: Secobarbital; Trimi Carteolol; Cefadroxil; Prednisolone: Reboxetine; Caspofun pramine; Cytarabine; Fondaparinux; Ofloxacin; Carbima gin; Nicotine; Gemcitabine; Pentostatin: Capecitabine; Vita Zole; Idoxuridine: Felbamate; Vitamin D3 (Cholecalciferol): min B6 (Pyridoxine); Leflunomide; Galantamine; Rifampin; Disopyramide; Terbinafine: Procainamide: Enalapril; Phen Metoprolol; Streptozocin; Metaproterenol; Crotamiton; Isof formin; Mephenytoin: Betamethasone; Metaxalone: Piren lurane: Cyclobenzaprine; Gentamicin; Morphine; Abacavir; Zepine: Fluorouracil; Sulfametopyrazine; Dolasetron; Amlo Torasemide: Pimozide. Sevoflurane; Naratriptan; Meman dipine; Daunorubicin; Proparacaine; Quinapril; Selegiline; tine; Buspirone; Olmesartan Medoxomil: Cevimeline; Pip Fosinopril; Diclofenac; Isosorbide Mononitrate; Meloxicam; erazine; Emtricitabine; Amitriptyline; Phenprocoumon; Fluoxetine: Apomorphine: Trazodone: Modafinil; Profla Timolol; Suprofen; Ibandronate; Netilmicin; Glyburide: vine; Vitamin B3 (Niacin); Ipratropium; Haloperidol; Ben Enflurane; Levothyroxime; Paramethadione; Topiramate: Zocaine; Ziprasidone; Ritodrine; Voriconazole; Chlorhexi Etoposide; Didanosine; Phenytoin: Mexiletine; Cefaclor; dine: Rosuvastatin: Minocycline: Propoxyphene: Primidone: Clotrimazole; Betaxolol; Calcitriol; Bupivacaine: Amoxicil Amikacin; Baclofen; Vitamin B1 (Thiamine); Albuterol: lin; Mechlorethamine; Cephalexin; Ethacrynic acid; Metaraminol; Sildenafil; Temozolomide: Nitazoxanide; Acetaminophen; Clomipramine: Ranitidine; Orlistat; Valp Marimastat; Lisinopril; Alendronate; Zalcitabine; Quinine; roic Acid; Bisoprolol; Trimeprazine: Paclitaxel; Cladribine; Beclomethasone; Lymecycline; Clindamycin; Acyclovir, Propafenone; Phenmetrazine; Ganciclovir, Aspirin; Zileu Cimetidine, Norgestimate; Lamotrigine; Marinol; Tetracy ton; Butalbital; Tolbutamide; Trimetrexate; Picrotoxin; Fro cline; Pemetrexed: Loxapine; Sotradecol; Dorzolamide: Vatriptan; Ridogrel, Demeclocycline; Enprofylline; Lopera Dexmedetomidine; Irinotecan; Alosetron: Tobramycin; US 2015/0309021 A1 Oct. 29, 2015

Cefixime: Astemizole; Diphenhydramine; Estrone; Terbuta Chlormerodrin; Chlormezanone; Chloroxine; Chlorphen line; Nifedipine; Hydrocodone; Aminoglutethimide; Nateg esin: CiclopiroX; Cinacalcet; Cinoxacin; Cisatracurium linide; Fludarabine; Sulfisoxazole; Thioridazine; Doxycy Besylate; Clemastine; Clobetasol; Clocortolone; Clofara cline; Halothane; Pyrimethamine; Famciclovir; bine; Clofibrate; Clomifene; Clorazepate; Cloxacillin; Promethazine; Nortriptyline: Moclobemide: Primaquine; Colesevelam: Colestipol; Colistimethate: Colistin; Crypte Amprenavir, Terfenadine: Hyoscyamine; Furosemide: Flu namine; Cyclacillin; Cyclopentolate; Cycloserine; cloxacillin; Mesoridazine; Nimodipine; Encainide; Atomox Cyclothiazide: Cysteamine Bitartrate; Dantrolene; Dapipra etine; Phentolamine; Scopolamine; Nicergoline; Framycetin: Zole; Darifenacin; Deferoxamine; Demecarium bromide: EZetimibe: Sulfinpyrazone; Bupropion; Bromocriptine: Deserpidine; Desflurane; Deslanoside; Desoximetasone: Saquinavir. Prochlorperazine; Estramustine; Vitamin B2 (Ri Dextrothyroxine; Dezocine; Diatrizoate; Diazoxide: boflavin); Medroxyprogesterone; Papaverine; BenZonatate; Dibucaine; Dicloxacillin; Dienestrol; Diethylcarbamazine: Cetirizine; Metronidazole; Finasteride: Fluphenazine: Pseu Diethylpropion; Diethylstilbestrol; Diflorasone; Diflunisal; doephedrine; Nisoldipine: Lisuride: Cinalukast; Aripipra Dimenhydrinate; Dimethyl Sulfoxide: Dinoprost Zole; Clodronate; Testolactone; Formoterol: Diazepam; Cef Tromethamine; Dinoprostone; Diphemanil Methylsulfate: dinir: Tranylcypromine: Penicillin G. Mimosine: Diphenidol; Dipivefrin; Dirithromycin; Docetaxel; Dexfenfluramine; Teniposide; Procaine; Phenoxyben Docosanol; Doxacurium Chloride; Doxapram; Doxylamine: Zamine; Altretamine: Pioglitazone; Fulvestrant; Dex Dromostanolone; Droperidol; Dyclonine; Dydrogesterone: tromethorphan; Acarbose; Methylphenidate; Terconazole; Echothiophate Iodide; Econazole; Edrophonium; Eletriptan; Thiopental; Acamprosate; Valrubicin; Pergolide; Busulfan; Emedastine; Enoxacin; Entecavir, Epoprostenol; Eprosartan; Metoclopramide; Bendroflumethiazide; Terazosin; Metipra Erlotinib. Ertapenem; Erythromycin; Esmolol; Estazolam: nolol; Foscarnet; Buprenorphine; Sufentanil; Imipramine; Ethambutol; Ethchlorvynol; Ethinamate; Ethiodol; Ethiona Caffeine; Dexamethasone; Quetiapine; Temazepam, Ergota mide; Ethotoin: Etidronate; Etomidate; Etretinate; mine; Pindolol; Norethindrone; Midazolam; Lamivudine; Fenoldopam; Fenoprofen; Flavoxate: Flumazenil; Flumetha Chlordiazepoxide; Escitalopram; Sulfamethizole; Mirtazap sone Pivalate: Flunisolide; Fluocinolone Acetonide; Fluoci ine; Cetrorelix; Topotecan; Hydroxyzine; Tripelennamine: nonide: Fluorescein; Fluorometholone: Fluoxymesterone: Tacrine; Ethinyl Estradiol; Floxuridine; Pipobroman; Novo Flurandrenolide: Flurazepam; Flutamide: Fomepizole; biocin; Procarbazine; Decamethonium; Valacyclovir; Leuco Fomivirsen; Fosfomycin; Furazolidone; Gadobenate dime vorin; Vardenafil; Progesterone: Isocarboxazid; Cerulenin: glumine; Gadodiamide; Gadopentetate dimeglumine; Gadot Sulfoxone: Nevirapine: Nizatidine: Eplerenone; Vinblastine: eridol: Gadoversetamide; Galsulfase; Ganirelix; Gatifloxa Desoxycorticosterone Pivalate; Bromodiphenhydramine: cin; Gefitinib; Gentian Violet; Glatiramer Acetate; Ergoloid Mesylate: Gallamine Triethiodide; Methdilazine: Glycopyrrolate: Gonadorelin; Granisetron; Grepafloxacin: Betazole; Chlorotrianisene; Chlorprothixene; Diphenylpyra Griseofulvin; Guaifenesin; Guanadrel Sulfate; Guanethidine; line; Chlorothiazide: Hexachlorophene; Buclizine; Adina Guanidine: Halazepam; Halobetasol Propionate; Halopro Zolam; Biperiden: Alfentanil; Bepridil; Benzthiazide; Ethop gin: Hetacillin: Hexafluorenium Bromide: Hexylcaine: His ropazine; Mefenamic acid; Cycrimine; EthoXZolamide; tamine Phosphate: Homatropine Methylbromide; Hydrocor Levallorphan; Methyprylon; Minaprine; Alprenolol; Clid tamate; Hydrocortisone; Hydroflumethiazide: inium: Ethanol; Methylergonovine; Methazolamide: Anileri Hydromorphone; Hydroxocobalamin: Hydroxypropyl Cellu dine; Melatonin; Methoxyflurane; Levomethadyl Acetate; lose; Hydroxystilbamidine Isethionate; Ibutilide Fumarate; Maprotiline; Benztropine; Aminosalicylic Acid; Isoetharine; Icodextrin, Iloprost: Imatinib. Imiquimod, Indapamide; Methantheline; Butabarbital; Flurbiprofen; L-Norgestrel; Indecainide; Inulin; Iron Dextran; Isoniazid: Ivermectin; Fludrocortisone; Metharbital; BenZphetamine; Ethynodiol Ketoprofen; Ketorolac. Ketotifen Fumarate; Labetalol; Diacetate; Dicumarol: Desogestrel; Isoflurophate; Levorpha Lactulose; Latanoprost: Letrozole; Levamisole; Levetirac nol; Carbinoxamine; Etonogestrel; Disulfiram; Dyphylline; etam; Levocabastine; Levocarnitine; Lindane; Linezolid; Dexbrompheniramine; Ambenonium: Acebutolol; Aceto Liothyronine; Lomefloxacin; Loracarbef; Loteprednol Eta hexamide; Acetohydroxamic Acid, Acitretin: Adapalene; bonate; Magnesium Sulfate; Malathion; Mannitol; Masopro Adefovir Dipivoxil; Albendazole; Alclometasone; Alpros col; Mazindol; Mebendazole; Meclofenamate; Medrysone: tadil; Alseroxylon; Amantadine; Amcinonide; Amdinocillin; Megestrol; Mepivacaine; Meprobamate; Mercaptopurine; Amiloride; Aminocaproic Acid; Aminohippurate; Aminole Meropenem; Mesalamine; Metformin; Metixene; Methocar Vulinic Acid, Amlexanox: Amodiaquine; Amphotericin B; bamol; Methyclothiazide; Methyl aminolevulinate; Methyl Anagrelide; Anisindione: Anisotropine Methylbromide: Arb dopa; Metolazone; Metyrapone: Metyrosine: Mezlocillin; utamine; Ardeparin; Atazanavir, Atovaquone; Atracurium; Micafungin; Miconazole; Midodrine; Miglitol; Miglustat; Atropine; Auranofin, AZacitidine, AZatadine; Azathioprine; Mivacurium; Moexipril; Monobenzone: Moricizine: AZelaic Acid, AZelastine, AZlocillin; Aztreonam, Bacitracin; Nabilone; Nabumetone; Nafarelin; Nafcillin; Naftifine; Nal Oxybuprocaine; Bentiromide; Bentoquatam, BenZquina buphine; Nalidixic Acid; Naproxen; Natamycin; Nedocro mide: Benzyl Benzoate; Benzylpenicilloyl Polylysine: Bet mil: Nitisinone; Nitric Oxide: Nitroprusside; Nystatin: Osel anidine; Bimatoprost; Bitolterol Mesylate; Bortezomib; tamivir Phosphate: Oxacillin: Oxamniquine; Oxandrolone: Bosentan; Bretylium; Bromfenac; Brompheniramine; Buten Oxazepam. Oxiconazole Nitrate; Oxymetazoline; Pamidr afine; Butoconazole; Butorphanol Tartrate; Cabergoline; Cal onate; Paricalcitol; Pemoline: Penicillamine; Pentagastrin; cium Acetate; Calcium Chloride: Calcium Gluceptate; Can Pentazocine; Pentobarbital; Pentosan Polysulfate; Perflutren: dicidin; Capreomycin; Carbachol; Carbenicillin: Carboprost Perindopril Erbumine: Pimecrolimus: Piroxicam, Podofilox: Tromethamine; Carphenazine; Carprofen; Cefditoren Piv Polymyxin B Sulfate; Pralidoxime; Praziquantel; Prednicar oxil, Cefnmenoxime; Cefnetazole; Ceforanide; Cefotaxime; bate; Pregabalin: Propofol; Pyrazinamide: Rabeprazole; Cefotiam; Ce?piramide: Ceftazidime; Cephaloglycin; Ramelteon; Remifentanil; Rifapentine; Rifaximin; Rimanta Cephalothin; Cephapirin; Ceruletide; Chloramphenicol; dine, Rimexolone; Rocuronium; Sevelamer; Sotalol; Spar

US 2015/0309021 A1 Oct. 29, 2015 thophyll; L-Lysine; L-Methionine; N-Acetyl-D- propion; diethylstilbestrol; difenoxin; diflorasone diacetate; glucosamine; NADH: Nicotinic acid: L-Ornithine; digoxin; dinoprost tromethamine; dinoprostone; diphen L-Phenylalanine: Pyridoxal P: Aspartame; L-Proline; Phos methanil-methylsulfate/diphemanil; diphenoxylate; dipiv phatidylserine: Pyridoxal; Pyruvic acid; Retinoic acid: S-Ad efrin, docetaxel; dolfetilide; dolasetron; donepezil; dopamine; enosylmethionine: L-Serine. Succinic acid; Spermine; Tet dorzolamide: doxacurium: doxazosin, doxepin, doxercalcif rahydrofolic acid; L-Threonine: L-Tryptophan; L-Tyrosine; erol: doxorubicin\epirubicin; dromostanolone propionate; L-Valine; Vitamin E; and Vitamin K3. dronabinol; drospirenone; dulloxetine; dutasteride: dyclo 0081. Other chemicals which may be used with the present nine; dydrogesterone; echothiophate; efavirenz; eflornithine; invention include the following, along with any salts and free eletriptan; emedastine; emitricitabine; enalaprilat; enflurane; bases: abacavir, abarelix; acamprosate; acarbose; enfuVirtide; enoxacin; entacapone; entecavir, epinastine; acetophenazine; acetylcysteine; acetyldigitoxin; acitretin: eplerenone; epoprostenol; eprosartan; eptifibatide; erlotinib; acrisorcin; acyclovir, adapalene; adefovir dipivoxil; ertapenem; erythromycinestolate; esmolol; estradiol acetate; adenosine\vidarabine; alatrofloxacin; albendazole; alclom estramustine phosphate; estrone; estropipate; esZopiclone; etaSone; alendronate; alfentanil; alfuZosin; almotriptan; alos ethchlorvynol; ethopropazine; ethotoin: ethylestrenol; etron; alpha-tocopherol\vitamine; alprostadil; ambenonium; ethynodiol diacetate; etidronate; etodolac, etonogestrel; eto amcinonide; amdinocillin; amikacin; aminoglutethimide; poside phosphate, etretinate; exemestane; exenatide; aminolevulinic acid; aminosalicylic acid; amiodarone; eZetimibe; famciclovir; famotidine; felodipine; fexofena amlexanox; amlodipine; amoxapine; amoxicillin; dine; finasteride; flavoxate; floxuridine; fluconazole; amphetamine\dextroamphetamine; amplicillin; amprenavir, fludeoxyglucose f-18; flumazenil; flumethasone pivalate: amrinone\inamrinone; anagrelide; anastroZole; anidulafun flunisolide; fluocinolone acetonide; fluocinonide; fluo gin, anilleridine; anisindione; apomorphine; apraclonidine; rometholone acetate; fluoxymesterone; fluphenazine enan aprepitant; arbutamine; argatroban; aripiprazole; atazanavir, thate; fluprednisolone; flurandrenolide; flurbiprofen; fluta atomoxetine; atorvastatin; atracurium\cisatracuriurn; aura mide; fluvastatin: fluvoxamine; fondaparinux; formoterol; nofin, avobenzone; azatadine, azathioprine; azelastine; fosamprenavir, foscarnet; fosinopril; fosphenytoin: fro aZlocillin; aztreonam, bacampicillin; baclofen; balsalazide; Vatriptan; fulvestrant; furazolidone; gabapentin; gadoteridol; beclomethasone; benazepril; benoximate; bentiromide; ben gallamine triethiodide; ganciclovir Sodium; ganirelix; gati Zonatate; benzquinamide; betamethasone benzoate; floxacing gefitinib, gemcitabine; gemfibrozil; gemifloxacin; betaxolol\levobetaxolol; betazole; bethanechol; bethanidine; gemtuzumab ozogamicing glimepiride; glucagon; glucono bexarotene; bicalutamide: bimatoprost; biperiden; biso lactone; glutethimide; gonadorelin; goserelin; gramicidin; prolol; bitolterol; bivalirudin; bortezomib; bosentan; brety granisetron; grepafloxacin; guanadrel; guanethidine:guanfa lium; brimonidine: brinzolamide: bromfenac; bromocriptine: cine; halcinonide; halobetasol propionate; haloperidol brompheniramine\dexbrompheniramine; buclizine; budes decanoate; haloperidol lactate; haloprogin; hetacillin; onide; buprenorphine; bupropion; buspirone; buSulfan; hexafluorenium; hexocyclium; hydrochlorothiazide; hydro butenafine; butoconazole; butorphanol; cabergoline; calcife cortamate; hydroxyprogesterone caproate; hydroxyStilbami diol, calcipotriol/calcipotriene; calcitonin human; calcitriol: dine; hydroxy Zine pamoate; ibandronic-acid\ibandronate; candesartancilexetil, candicidin; capecitabine; capreomycin; ibuprofen lysine; ibutilide; icodextrin; idarubicin; idoxuri carbachol; carbenicillinindanyl sodium; carboprost, carmus dine; ifosfamide; iloprost; imatinib, imipenem; imipramine tine; carphenazine; carprofen; carteolol; carvedilol; caspo pamoate; imiquimod; indinavir, indocyanine green; ioben fungin, cefaclor, cefazolin, cefepime; cefotaxime; cefoxitin; guane; iocetamic acid; iodamide meglumine; iodipamide ceftazidime; cefuroxime; celecoxib; cephalexin; cephalogly meglumine; iodixanol; iodoxamate meglumine; iofetamine; cin; cephalothin; cephapirin; cephradine; cerivastatin: ceru iohexol; iopamidol; iophendylate; iopromide; iothalamate letide; cetirizine; cetrorelix; cevimeline; chenodiol\ursodiol; diagnostic; iotrolan; ioVersol; ioxaglate meglumine; ioxilan; chlorambucil; chloramphenicol palmitate; chlordiazepoxide; ipodate; irbesartan; irinotecan; isocarboxazid; isoetharine; chlorhexidine gluconate; chlorphenesin carbamate; isoflurane; isosorbide dinitrate; isosulfan blue; isradipine; chlorpheniramine\dexchlorpheniramine; chlortetracycline; ivermectin; ketotifen; labetalol; lamivudine; lamotrigine; chlorthalidone; cholecalciferol; ciclopirox: cidofovir; cilas latanoprost; leflunomide; lenalidomide; lepirudin; letrozole; tatin; cilostazol; cinacalcet; ciprofloxacin, cisapride; leuprolide; levacetylmethadol hydrochloride\levomethadyl citalopram\escitalopram; cladribine; clavulanate; clemas acetate; levallorphan; levamisole; levetiracetam; tine; clindamycin palmitate; clobetasol propionate; clocor levobunolol; levocabastine; levocarnitine; levomepromazine; tolone pivalate; clofarabine; clopidogrel; cloraZepate; cloX levonorgestrel\norgestrel; liothyronine\liotrix; lisinopril; acillin; clozapine; cobalamin\hydroxocobalamin; colfosceril lithium; lodoxamide; lomustine; lopinavir, loracarbef losar palmitate; colistimethate sodium; colistin; conivaptan; corti tan; loteprednol etabonate; lovastatin; loxapine; lubipros corelin ovine triflutate; corticotropin; cosyntropin; cyclacil tone; lypressin\vasopressin tannate; mazindol; mecamy lin; cyclobenzaprine; cyclopentolate; cyclosporine; lamine; meclocycline Sulfosalicylate; medrysone; megestrol cycrimine; cytarabine; dacarbazine, dactinomycin; dalfopris acetate; meglumine metrizoate; meloxicam, melphalan; mep tin, dantrolene; dapiprazole; daptomycin; darifenacin; eridine; mephentermine; mephenytoin: meprednisone; mer deferasiroX, delavirdine; demecarium; desirudin; deslano captopurine; meropenem; mesna; mesoridazine; metaxalone; side; desloratadine; desmopressin; desogestrel; desonide; methacycline; methanthelinium\methantheline; methaZola desoximetasone; desoxycorticosterone pivalate; dexametha mide; methdilazine; methenamine hippurate; methicillin; SO Sodium phosphate; dexmedetomidine; methimazole; methixene; methohexital; methoxamine; dexmethylphenidate\methylphenidate; dexpanthenol; dexra methoXSalen; methSuximide; methylaminolevulinate; meth ZOXane, dextromethorphan polistirex; yldopate; methylergonovine; methylprednisolone acetate; dextrothyroxine\levothyroxine; dezocine; diatrizoate meglu methyprylon; methysergide; metipranolol; metocurine; mine; dichlorphenamide; dicloxacillin; didanosine; diethyl metolaZone; metrizoate\metrizoic acid; metronidazole; US 2015/0309021 A1 Oct. 29, 2015 36 metyrapone; metyrosine; mezlocillin; micafungin; mido coxib; Valganciclovir, Valproate\valproic acid\divalproex; drine; mifepristone; miglitol; miglustat; milrinone; minoxi valrubicin; Valsartan; Vardenafil; vecuronium; Venlafaxine; dil; mirtazapine; misoprostol; mitotane; mitoxantrone; miva Verapamil: verteporfin; Vinorelbine; viomycin; vitamin a curium: moexipril; molindone; mometasone furoate; palmitate; Voriconazole; Zaleplon; Zanamivir, Ziconotide; montelukast, moricizine; moxalactam; moxifloxacin; mupi Zidovudine; Zileuton, Ziprasidone; Zoledronic acid; Zolmi rocin, nabilone; nafarelin, naftifine; nalbuphine; nalmefene; triptan; Zolpidem; Zonisamide, 1-alpha-24-dihydroxycolecal naloxone; naltrexone; naratriptan; natamycin; nateglinide; ciferol; 2',3'-dideoxythymidine; 5-hydroxyrofecoxib; nedocromil; nefazodone; nelarabine; nelfinavir, neomycin; acecainide-hydrochloride; aceclidine; aceclofenac; acegla nepafenac; nesiritide; netilmicin; nevirapine; nicardipine; tone; acenocoumarol; acepromazine-maleate; acetaminos niclosamide; nifedipine; nilutamide; nimodipine; nisol alol; acetiamine; acetosulfone sodium; acetoxolone; acetyl dipine; nitaZoxanide; initisinone; nitrofurantoin: nitrofura carnitine; acetyldigoxin-beta; acetyldihydrocodeine: Zone; nitroglycerin; nizatidine; norelgestromin; norethin acetylstrophanthidin; acetylsulfamethoxypyrazine; acipi drone acetate; norethynodrel; norgestimate; nystatin: moX, acivicin; aclarubicin; ademetionine-disulfate-ditosi octinoxate; octreotide: olanzapine; olmesartan medoxomil: late; adenine hydrochloride, adrafinil; afloqualone; ag-81: olopatadine; omega-3-acid ethyl esters; omeprazolefesome ajmalicine; aklomide; alacepril; alafosfalin; alanosine; ala prazole; ondansetron; orlistat; orphenadrine citrate; oselta proclate; alclofenac; alcloxa: alcuronium-chloride; aldes mivir; oxacillin, oxamniquine; Oxandrolone; oxaprozin, ulfone; aldioxa; alfacalcidol; alfadolone-acetate; alfaprostol; oXcarbazepine; oxiconazole; OXtriphylline; palonosetron; alfaxalone; algestone-acetophenide; alibendol; alinidine; pamidronate; pancuronium; pantoprazole; pantothenic acid; alizapride; alloclamide; alloyohimbine; allylestrenol; almi paramethadione; paramethasone acetate; pargyline; parical noprofen; almitrine; aloin, alpha-acetyldigoxin; alphaprod citol; paroxetine; pegaptanib; pemetrexed; pemirolast; penb ine-hydrochloride; alprenolol-benzoate; alrestatin-Sodium; utolol; penciclovir, penicilling procaine; pentagastrin; pen alsactide; aluminum-clofibrate; alverine-citrate; ambicromil: tamidine; pentazocine; pentetreotide; pentolinium; ambucetamide; ambuside; ambutonium-bromide; amdinocil pentostatin: pergolide; perindopril; permethrin; perphena lin-pivoxil, amezinium-metilsulfate; amfenac; amfetaminil; Zine; phensuximide; phentolamine; phenyl aminosalicylate; amifloxacin; amineptine; aminorex, aminothiazoline cam phenylbutaZone; pimecrolimus; pinacidil; pindolol; pioglita phorate; amiprilose-hydrochloride; amisulpride; amitrip Zone; pipecuronium; piperacetazine; piperazine; pipobro tyline embonate; ammonium peroxydisulfate; amorolfine man; pirbuterol, piroXicam, plicamycin; polyestradiol phos hydrochloride; amoSulalol hydrochloride; amphomycin; phate; polymyxin b; porfimer; povidone-iodine: amprolium; amylmetacresol; androsterone; anetholtrithion; pramipexole; pramlintide; pravastatin; prednicarbate; pred angiotensin-2; angiotensin-amide; antrafenine; apalcillin; nisolone acetate; pregabalin; procarbazine; prochlorperazine apaZone: aprindine-hydrochloride; arabinosyluracil-triphos edisilate; promethazine; propiolactone; propiomazine; pro phate; arbaprostil, argipressin, armodafinilvmodafinil; aroti poxyphene; propoxyphene napsylate; propylthiouracil; pro nolol, arsphenamine; aspoxicillin; atosiban; atromepine; tirelin; protriptyline; pseudoephedrine; pyrvinium pamoate; avoparcin; azamethonium-bromide; azanidazole; azaperone; quetiapine; quinapril; quinestrol; quinupristin; rabeprazole; aZapetine-phosphate; azaribine; azauridine; azidamfenicol; raloxifene; ramelteon; ramipril; ranolazine; rapacuronium; azidocillin; azolimine; aZosemide; bacitracin; bambermycin; remifentanil; repaglinide; rescinnamine; rifapentine; rifaxi bambuterol-hydrochloride; bamethan; bamifylline-hydro min, riluzole, rimantadine; rimexolone; risedronate; risperi chloride; bamipine; barbetonium iodide; barbexaclone: done; ritodrine; ritonavir, rivastigmine; rizatriptan; rocuro befunolol; befuraline hydrochloride; bekanamycin; bemetiz nium; rofecoxib; ropinirole; rosiglitaZone; rosuvastatin: ide; benapry Zine; bencyclane; bendaZac; benethamine-peni salmeterol Xinafoate, saquinavir, Saralasin; Secretin; sel cillin; benfluorex; benfotiamine; benfurodil-hemisuccinate; egiline; selenomethionine; seractide; sermorelin; Sertacona benorilate; benoxaprofen; benperidol; benproperine Zole; sertraline; sevoflurane; sibutramine; sildenafil citrate; embonate; benzalamide; benzaldehyde; benzarone; benzilo simvastatin; sincalide; Sirolimus; Solifenacin; Sorafenib; nium-bromide; benziodarone; benznidazole; benzoctamine; Sotalol, spirapril; stanozolol, stavudine; streptomycin Sulfate; benzopiperylone; benzquercin; benzydamine Salicylate; ben streptozocin, Sufentanil; Sulbactam, Sulfacytine; Sulfamera Zylhydrochlorothiazide; bergapten; betamipron; bevantolol; Zine; sulfameter, Sulfamethazine; Sulfisoxazole acetyl: bevonium-metilsulfate; bezitramide: bibrocathol; bicloty Sumatriptan; Sunitinib. Supprelin/histrelin; Suprofen; tacroli mol; bietaserpine; bifemelane hydrochloride; binifibrate; mus; tadalafil; tamoxifen, tamsulosin; tannic acid; tazaro biphenamine-hydrochloride; bisantrene; bisbentiamine; bis tene; taZobactam, tegaserod; telmisartan; temozolomide; butitiamine; bisdequalinium diacetate; bisorcic, bisoxatin teniposide; tenofovir disoproxil; terazosin; terbinafine; teri acetate; bm-23014; bolasterone; boldenome-undecylenate: paratide; testolactone; testosterone; tetrahydrozoline; thali bornaprine; brivudine: bromhexine; bromperidol-decanoate: domide; thiethylperazine; thioguanine; thiopental; thiothix bronopol; broparestrol; brotianide; broxaldine; broxuridine: ene; thonzonium; tiagabine; ticarcillin; tigecycline; bucindolol; buclosamide; bucloxic-acid; bucolome; bufe tiludronate; timolol; tinidazole; tioconazole; tiotropium; tolol; bufotenine; bufrolin; bufuralol; bumadizone; bunami tipranavir; tirofiban; tizanidine; tobramycin; tolcapone; dine; bunazosin hydrochloride; bunitrolol; bupranolol; tolterodine; topiramate; toremifene; torsemide; trandolapril; buquineran; buserelin; buserelin-acetate; butalamine; tranylcypromine; travoprost; treprostinil; butamirate; buthiazide; butibufen, butikacin; butofilolol; tretinoin\alitretinoin\isotretinoin, triamcinolone; triclofos; butopiprine; butriptyline; butropium-bromide; buzepide-me tridihexethyl; triflupromazine; trifluridine; trilostane; tri thiodide; cadralazine; cafaminol; caffeine sodium benzoate; methaphan camsylate; trimethobenzamide; trimipramine; tri camo.stat mesilate; camylofin, canrenoate-potassium; can oXSalen; triptorelin pamoate; troglitaZone; tropicamide; tro renone; canthaxanthin; capobenic-acid; captodiame-hydro spium; trovafloxacin; tubocurarine; tyloxapol; tyropanoate; chloride; carazolol, carbadox; carbazochrome; carbimazole; undecoylium; unoprostone isopropyl; Valacyclovir, Valde carbinoxamine-tannate; carbomer; carboprost-methyl; car US 2015/0309021 A1 Oct. 29, 2015 37 boduone; carbutamide; carbuterol-hydrochloride; carglumic diphenadione; diphenan; diphenhydramine acefylline; diphe acid; carnidazole; caroverine; carpipramine; carubicin; caru nylthiocarbazone; dipipanone-hydrochloride; diprenorphine; monam-Sodium; celiprolol-hydrochloride; cephacetrile dipyrocetyl; disodium adenosine-triphosphate; disulfamide; Sodium; cephaloridine, cethexonium-chloride; cetiedil; ditazole; dithranol-triacetate; dixyrazine; dodicin; domiodol; cetiedil citrate; cetostearyl-alcohol; cetotiamine; cetraxate domperidone; dopexamine, doxantrazole; doXpicomine-hy hydrochloride; cetylpyridinium-chloride; chloral-betaine; drochloride; dropropizine; drostanolone\dromoStanolone; chloramine-b; chlorbenzoxamine; chlorcyclizine; chlorhexa drotaverine; drotebanol; echothiopate-iodide; edoxudine: dol; chlormadinone-acetate; chlormethiazole; chlormethine efaproxiral; efloxate, elcatonin; eledoisin; elizabethin; ellip n-oxide hydrochloride; chlormidazole; chloroxylenol; tinium-acetate; embramine; emepronium-carrageenate; chloroxymorphamine; chlorozotocin, chlorphenoxamine endralazine-mesylate; endrysone; enilconazole; enoXimone; embonate; chlorproethazine-hydrochloride; chlorpromazine enprostil; enviomycin; enviroxime; eperisone-hydrochlo embonate; chlorpyrifos, chlorquinaldol; chlorthenoxazine; ride; epicillin; epiestriol; epimestrol; epithiazide; epi cholesterol: cholinemagnesium trisalicylate; chromocarb; tioStanol; epomediol, eprazinone; eproZinol; eptazocine; chromonar, chrysarobin; cianidanol: ciclaZindol; erythrityl-tetranitrate; erythromycin-propionate; erythromy ciclesonide; cicletanine: ciclonicate; ciclosidomine; ciclox cin-Succinate, erythrosine; eseridine; etamiphyllin camsilate; ilic-acid; cilastatin, n-acetyl; cilaZapril; cilobamine-mesy etebenecid; eterobarb; etersalate; ethadione; ethaverine; ethi late; cinametic-acid; cinchophen; cinepazet, cinepazide; cin azide; ethopabate; ethyl-biscoumacetate; ethyl-icosapentate; metacin; cinoxate, cinpropazide maleate; ciprofibrate; ethyl-loflazepate; ethyl-Orthoformate; ethynodiol; etifelm ciprostene-calcium; ciramadol-hydrochloride; citicoline ine; etifoxine; etiroXate: etodroxizine; etofamide: etofe sodium; clebopride; clefamide; clemizole sulfate; clen namate; etofibrate; etoglucid: etoperidone-hydrochloride; buterol; clidanac; clinofibrate; clioxanide; clobazam; cloben etoricoxib: etosalamide; exalamide; exifone; exiproben; Zorex; clobetasol; clobetasone-butyrate; clobutinol: exisulind; fampiridine; famprofaZone; faZadinium-bromide; clobuZarit; clocanfamide; clocapramine; clocinizine; clo febantel; febuprol; feclobuzone; fedrilate; fely pressin; fenaf dronic acid; clofarabine-triphosphate; clofenamide; clofexa tic-acid; femalamide; femalcomine; fenbendazole; fen mide; clofibrate pyridoxine; clofibride; clofilium-phosphate: butrazate; fencamfamin; fencibutirol; fenclofenac; fenclo clofoctol; cloforex; clometacin; clomethiazole edisilate: nine; fenclozic-acid; fendosal; fenfluramine hydrochloride; clometocillin; clomocycline; clonixin; clopamide; clo fenitrothion; fenmetozole; fenoterol; fenoverine; fenoxazo penthixol-decanoate; cloperastine fendizoate; clopirac, clo line-hydrochloride; fenozolone; fenpentadiol; femproporex: prednol; cloprostenol-sodium; cloranolol; clorexolone; clor fenprostalene; fenduizone; fenretinide; fenspiride; fenthion; giline; cloridarol; clorindione; clorophene; clorprenaline fentiazac, fenticonazole; fentonium-bromide; feprazone; hydrochloride; clortermine; clostebol-acetate; clothiapine; fezatione: fibracillin; flavodic-acid; floctafenine; floranty clovoxamine; cloxacillin benzathine; cobamamide; cocar rone; floseduinan; fluianisone; fluazacort; flucloronide; flu boxylase magnesium; colestyramine hydrochloride; colext darabine; fludrocortisone; flufenamate-decanoate; fluindi ran; colimycin; colistin Sulfate; colistin-b; congo-red; conor one; flumedroxone-acetate; flumequine; flumeridone; phone-hydrochloride; cortivazol; cotarnine-chloride; flumethiazide; flunixin-meglumine; flunoxaprofen; fluocor coumazoline; coumetarol; creatinolfosfate; croconazole tin-butyl; fluocortolone; fluorescein-dilaurate; fluoresone; hydrochloride; cropropamide; cyamemazine; cyclandelate; fluotracen; flupamesone; flupentixol; flupentixol-decanoate; cyclazocine; cycloartenol-ferulate; cyclobendazole; cyclofe fluperolone-acetate; flupirtine; fluprednidene-acetate; flu nil, cycloguanil-pamoate; cyclopentamine-hydrochloride; prostenol-sodium; flurothyl; fluspirilene; fluticasone propi cyclopenthiazide; cycotiamine; cynarine; cyprodenate; onate; flutroline; fominoben; fonazine-mesylate; forme cyproterone, cyproterone-acetate; cytochalasine-d; cytosine; bolone; formestane; formocortal; formosulfathiazole; danitracen; deanol-aceglumate; deanol-phosphate; debriso fosfosal; furapromidium; furonazide; fursultiamine; fusaric quin; decoquinate; deflazacort, dehydroemetine; delapril acid, gabexate mesilate; gabob; gallopamil, gamolenic-acid; hydrochloride; delmadinone-acetate; demegestone; demex ganglioside; gefarnate, gemeprost, gepefrine; gepirone-hy iptiline; demoxytocin, denatonium-benzoate; deoxyglucose: drochloride; gestodene; gestrinone; gitaloxin; gitoformate; deptropine-citrate; desaspidin; detorubicin; dexecadotril; glaucine phosphate; glibornuride; gliflumide; gliquidone; dexetozoline; dexlofexidine; dextranomer; dextrofemine; glisentide; glisolamide; glisoxepide; glucametacin; glucosul dextromoramide; diacerein; diacetolol-hydrochloride: fone; glucuronamide; glutathione; glybuzole; glyclopyra diacetylmorphine-hydrochloride; diamfenetide; dianhydro mide; glyconiazide; glycyclamide; glymidine-Sodium; galactitol; diaveridine: dibekacin: dibenzepin; dibenzoyl-dis gramicidin-S, guabenXan; guacetisal; guaiacol-phenylac ulfide; dibrompropamidine: dibromsalan: dibunate-sodium; etate; guaiapate; guaietolin; gualenate-sodium; guamecy dichloralphenazone; dichlorisone-acetate; dichlorodifluo cline; guanacline-Sulfate; guanaZodine Sulfate; guanoclor romethane; dichlorotetrafluoroethane; diclofenamide Sulfate; guanoxabenz, guanoxan; halofenate; halofluginone; sodium; diclofensine; dicolinium-iodide; dieldrin; diethyl halometaSone; halopredone; halopredone-acetate; haloxon; phthalate; diethyltoluamide; difemerine-hydrochloride; dife hematoporphyrin, hepronicate; hetacillin-potassium; namizole; difenpiramide; difetarsone; diflucortolone-valer hexachloroethane; hexacyprone; hexamethonium-tartrate; ate; difluprednate; diftalone; dihexyverine-hydrochloride: hexamidine; hexapropymate; hexestrol-dipropionate; hexeti dihydralazine; dihydroergocryptine-beta mesilate; dihydro dine; hexobendine; hexoprenaline; histapyrrodine; tachysterol; dihydroxydibutylether; diisopromine; dilazep: homidium-bromide; homofenazine; hopantenate calcium; diloxanide; dimecrotic-acid; dimefline-hydrochloride: hyaluronate-sodium; hycanthone; hydrastine-hydrochloride; dimenorfan; dimethindene maleate; dimethisterone; dim hydrobentizide, hydroxydione-sodium-Succinate; hydrox ethylacetamide; dimethyl-phthalate; dimethyltryptamine: yestrone-diacetate; hydroxyphenamate; hypoglycin-a; ibac dimetofrine; dimetridazole; dimoxyline phosphate; dini itabine; ibopamine; ibudilast; ibufenac; ibuproxam: ibuterol: profylline; dioxation; dioxybenzone; dipenine-bromide; idebenone; idrocilamide; ifenprodil; imafen-hydrochloride: US 2015/0309021 A1 Oct. 29, 2015 imaZodan; imcarbofos; imidapril; imidocarb; imipraminox oxymesterone; oxypendyl, oxypertine; padimate-a; padi ide; imolamine; impromidine-hydrochloride; indeloxazine; mate-o; palmidrol; pancreozymin, panipenem; papaveroline; indenolol; indobufen, indoramin; inosine-pranobeX; ioben parapenzolate-bromide; pararosaniline-pamoate; parathiaz Zamate; iocarmic acid; iodamide; iodopyriacet (1125); iodot ine teoclate; parecoxib; pargeverine; parsalmide; pas-hy hiouracil; iodothymol; iodoxamic-acid; iotasul; ipriflavone; drazide; pasiniazid; pecilocin, pefloxacin; penfluridol; pen iproclozide, ipronidazole; isoaminile; isobromindione; gitoxin; penimepicycline; pentaerythritol-tetranitrate; isobutiacilate; isoconazole; isoflupredone-acetate; isoleu pentagestrone-acetate; pentetate meglumine; penthienate; cine; isoly Sergide, isomalt, isonixin; isospaglumate magne pentisomicin; pentrinitrol; peplomycin-sulfate; pepstatin: sium; isosulphan-blue; josamycin-propionate; kainic-acid; perazine; perhexiline phosphate; periciazine embonate; per kawain; kebuZone; keracyanin; ketanserin; ketobemidone; lapine; peruvoside; pheneticillin; phenglutarimide: phen kitasamycin; lachesine-chloride; lacidipine; lactalfate; lacti probamate; phenyltoloxamine citrate; phenyramidol; pholco tol; laetrile; lanatoside-c; lasalocid-Sodium; laurolinium dine, phthalofyne; phytate-persodium; picloxydine; acetate; lawsone; lercanidipine; lergotrile-mesylate; leto picotamide, pifarnine; pimethixene; pinaverium-bromide; Steine; leucocianidol; levomepromazine embonate; levo pipamazine; pipamperone; pipazethate; pipebuZone; piper moprolol; levopropoxyphene dibudinate; levopropylcillin; oxan; pipethanate-ethobromide; pipotiazine-palmitate; levopropylhexedrine; levoprotiline; levosimendan; lidofla pipoxolan-hydrochloride; pipratecol; piprinhydrinate; pipro zine; lisuride; lobeline; lodoxamide, desethyl: lofentanil Zolin; pirarubicin; piraZolac, pirenoXine;piremperone; piren oxalate; lofepramine; loflucarban; lonazolac, lonidamine; Zepine; piretanide; piribedil: pirifibrate; piriMiphos-ethyl: lorajmine-hydrochloride; lornoxicam, lucanthone; lupros pirimiphos-methyl; pirisudanol; piritramide; pirlindole; pir tiol; lymecycline; lynestrenol; magaldrate; magnesium glu menol-hydrochloride; piroheptine; piromidic-acid; piroxi ceptate; malachite-green-oxalate; mandelic acid; mannomus mone: pirozadil; pirprofen; pivampicillin; pivoefalexin tine; maZaticol; mebanazine; mebeverine embonate; hydrochloride: pizotyline; plafibride; plaunotol; poldine: mebhydrolin; mebhydrolin napadisilate; mebolazine; mebu polymyxin; polynoxylin; polyoxyethylene-cetyl-ether; tizide, mecobalamin; mecrifurone; medifoxamine; medroge polyphloroglucinol-phosphate; ponalrestat; potassium poly stone; medroxalol hydrochloride; medroxyprogesterone; styrene-Sulfonate; practolol; prajmalium bitartrate; pramiver medrylamine; mefenidramium-metilsulfate; mefenorex; ine; pranoprofen; pranosal; prednazoline; prednimustine; mefruside; meglutol; melanostatin: melarsonyl-potassium; prednisolamate; prednylidene; prednylidene-diethylami melitracen; melperone; memotine-hydrochloride; menate noacetate; prenalterol-hydrochloride; prenoxdiazine; trenone; mepartricin-lauryl-sulfate sodium; mephenoxalone: prenoxdiazine hibenzate; prenylamine; pristinamycin; pro mepindolol; mepitioStane; mepixanox; meproscillarin; caterol; procodazole; profenamine hibenzate; progabide; meptazinol; mesterolone; mesulergine; mesulfen, metabro proglumetacin; proglumide; promazine embonate; pro msalan; metadoxine; metahexanamide; metallibure zinc: mazine-hydrochloride; promegestone; promestriene; propac metapramine; meteneprost, metenolone-enanthate; meter etamol; propallylonal; propamidine; propanidid; propatyl goline; metescufylline; metformin embonate; methallenes nitrate; propiram; propiverine hydrochloride; propizepine; tril; methaniazide; methenolone-acetate; methoprene; meth propyleneglycol-monostearate; propyl-gallate; propy oserpidine; methoxyphenamine; methylergometrine; phenaZone; propyromazine-bromide; produaZone; pro methylthiouracil; metiazinic-acid; metindizate; metioprim; renoate-potassium; proscillaridin; prostalene; prothipendyl; metopimazine; mevastatin; mianserin; mibefradil dihydro protionamide; protizinic-acid; protoveratrine-b; proxazole; chloride; mibolerone: mikamycin; milnacipran; miloxacin; proXicromil, prozapine; psilocybine; puromycin; pyrantel; misonidazole; mizolastine; mocimycin; moclobemide; pyrantel-pamoate; pyricarbate; pyritidium; pyritinol; pyrov mofebutaZone; molsidomine; moperone; mopidamol; mor alerone-hydrochloride; pyrrobutamine-phosphate; pyrrolni antel-tartrate; moraZone; morclofone; morinamide; morni trin; quinagolide; quinbolone; quinestradol; quingestanol-ac flumate; motretinide; moXaverine; moxestrol; moxonidine; etate; quinidine Sulfate; quinocide; quinoline-yellow: muZolimine; naboctate-hydrochloride; nadide; nadoxolol; quinupramine; quinuronium-sulfate; rafoxanide; raltitrexed: nafarelin; nafronyl: naftalofos; naftaZone; nalorphine; nan ramoplanin; ranimustine; rasagiline mesilate; reboxetine; drolone-cyclohexylpropionate; nebivolol; nequinate; neu remoxipride; reproterol: rifamide: rimazolium-metilsulfate; tral-red; niaprazine; nicametate; nicarbazin; nicergoline; nic rimiterol: rioprostil; ritanserin; rociverine; rolipram, ronida eritrol; niclofolan; nicoclonate, nicofibrate; nicofuranose; Zole; rosaprostol; rosaramicin; roSoxacin; roXarsone; roXati nicofurate; nicorandil; nifemalol; nifuratel; nifurfoline; dine acetate; roXithromycin, ruscogenin; Safrazine; salicy nifuroxazide; nifursol; nifurtimox; nifurtoinol; nifurzide; late-ethyl; salmefamol; sapropterin; so-w 44463: nikometamide; nilvadipine; nimeSulide; nimorazole; nimus secnidazole; Sefavirenz; Semustine; Sennoside; Sennoside-a; tine; niridazole; nisin; nithiamide; nitrefazole; nitrendipine; serotonin; sertindole; simfibrate; sizofiran; Sobrerol; sodium nitroscanate; nitrovin; nitroXinil; nomegestrol-acetate; cefuZonam, Sodium estriol-succinate; sodium flomoxef. nomifensine; nonivamide; norbormide; nordefrin-hydrochlo Sodium picosulfate; sodium polymetaphosphate; sodium ride; norethandrolone; norethisterone-enanthate; norg prednisolone-meta-sulfobenzoate; sodium warfarin, Sodium estrienone; noscapine; nosiheptide; noxiptiline; octafonium bensuldazate, sodium-loxoprofen; sodium-OZagrel, Solasul chloride; octanoic-acid; Octotiamine; octoxynol-9, fone; somatostatin; Sorbinil; Sorbitan-monolaurate; spaglu oleandomycin-phosphate; omapatrilat; ondaScora; mic-acid; Sparteine amp; Sparteine theophylline; opipramol hydrochloride; ornidazole; ornipressin: Oxab spiclomazine; spiperone; spiramycin adipate; stallimycin; olone-cipionate; oxafloZane; oxametacin: Oxantel-pamoate; stepronin; Stibocaptate; stilbophen; stilbazium iodide; stro oxapium-iodide; Oxatomide; oxendolone; oxetorone; phanthin-h; Succinimide; Succinylsulfathiazole bismuth, Sul oxfendazole; Oxfenicine; oxibendazole; oxidopamine; OXini benicillin; Sulbutiamine; Sulfadicramide; Sulfaguanole, Sul acate olamine; oxiperomide; oxiracetam; oxitriptan; oxybro famethoxypyridazine-acetyl: sulfametrole; monaftoic-acid; oxychlorosene; oxyclozanide; oxyfedrine; Sulfamonomethoxine, Sulfanitran; Sulfaperin; Sulfaproxy US 2015/0309021 A1 Oct. 29, 2015 39 line, Sulfaquinoxaline; sulfarsphenamine; Sulfazamet, Sul bromfenac, bromocriptine, bromodiphenhydramine, bro firam; Sulisatin: Sulmazole; Sulpiride adamantanecarboxy mpheniramine, buclizine, bumetanide, bupropion, buSulfan, late; Sulprostone; Sulthiame; Sultopride; Sultroponium; butalbital, butoconazole, capecitabine, capreomycin, capto Sunset-yellow; SuXethonium-chloride; SuXibuZone; Syrosin pril, carbenicillin, carbenicillin indanyl, carbinoxamine, car gopine; talampicillin-hydrochloride; taleranol; tamitinol; tar boplatin, carmustine, carphenazine, carprofen, cefaclor, trazine; taxol; teclothiazide, teclozan, tedelparin, tefazoline; cefadroxil, cefamandole, cefazolin, cefdinir, cefditoren, teicoplanin; temafloxacin; temocillin; temoporfin, tenamfe cefepime, cefixime, cefimenoxime, cefimetazole, cefonicid, tamine; tenidap; tenitramine; tenonitrozole; tenoxicam, cefoperaZone, ceforanide, cefotaxime, cefotetan, cefotiam, teprenone; teprotide; terizidone; terlipressin; terlipressin cefoxitin, ce?piramide, cefpodoxime proxetil, cefprozil, acetate; terodiline-hydrochloride; tertatolol; tetrabenazine: ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, tetradecylsulfate-sodium; tetrahydrozoline hydrochloride; cefuroxime, cefuroxime axetil, celecoxib, cephalexin, cepha tetramethrin, tetrydamine; tezosentan; thebacon; theobro loglycin, cephalothin, cephapirin, cephradine, cerivastatin, mine-sodium-acetate; theofibrate; theophylline-acetate-so dium; thiambutosine; thiamine-propyl-disulfide; thiam cetirizine, cetrorelix, cevimeline, chlophedianol, chloram phenicol-glycinate hydrochloride; thiamphenicol-palmitate; bucil, chloramphenicol, chlordiazepoxide, chlorhexidine, thiamylal-Sodium; thiazinamium; thiazolsulfone; thibenza chlormeZanone, chloroprocaine, chloroquine, chlorothiaz Zoline; thiocoichicoside; thiophanate; thiopropazate; thiou ide, chlorotrianisene, chloroxine, chlorphenesin, chlorphe racil; thymalfasin; thymopentin; tiamenidine; tiamulin-fu niramine, chlorphentermine, chlorpromazine, chlorpropam marate; tianeptine; tiapamil-hydrochloride; tiapride; ide, chlorprothixene, chlortetracycline, chlorthalidone, chlorZoxaZone, chromium picolinate, chromium salts, cilas tiaprofenic acid; tiaramide-hydrochloride; tibeZonium-io tatin, cimetidine, cinacalcet, ciprofloxacin, cisapride, cispl dide; tibolone; ticrynafen; tidiacic, tiemonium-iodide; tigloi atin, citalopram, cladribine, clemastine, clindamycin, clinda dine; tilidine-hydrochloride; timepidium bromide; timonacic mycin, clindamycin, clioquinol, clobetasol, clocortolone, arginine; tinoridine, tiocarlide; tioclomarol; tioXolone; tipe clofarabine, clofazimine, clofibrate, clomiphene, clomi pidine hibenzate; tiquizium-bromide; tiratricol; tirilazad; pramine, clonazepam, clonidine, clopidogrel, cloraZepate, tiropramide; tiXocortol-pivalate; tocamphyl; tocopherol-al clotrimazole, cloxacillin, clozapine, colestipol, cyclophos pha-nicotinate; tocopheryl-quinone; todralazine; tolciclate; phamide, cyclothiazide, cysteamine, cysteine, dalfopristin, tolmesoxide; tolnaftate; toloconium-metilsulfate; tolonidine; toloxatone; tolperisone; tolpropamine; tolrestat; tonazocine dapsone, delavirdine, demeclocycline, desflurane, deslorata mesylate; tosactide; tramazoline; tranilast; trenbolone; tren dine, desmopressin, dexamethasone, dexamethasone, dex bolone-cyclohexylacetate; trengestone; treosulfan; treto amethasone, dexbrompheniramine, dexchlorpheniramine, quinol; triacetin; tribenoside; tribromoethanol; tribuZone: diatrizoate, diazepam, diazoxide, dichlorphenamide, triclobisonium-chloride; triclocarban; triethylenemelamine; diclofenac, dicloxacillin, diethylstilbestrol, diflorasone, trifenmorph; trifluperidol; triflusal; triiodostearate-ethyl; tri diflunisal, diltiazem, dimenhydrinate, dimercaprol, dimethyl maZosin; trimebutine; trimetazidine; trimethidinium-metho sulfoxide, dimyristoyl, disulfiram, dofetilide, dorzolamide, sulfate; trimetrexate; trimoprostil; tripamide; tritiozine; trito droperidol, duloxetine, dutasteride, echothiophate, econa qualine; trofosfamide; trolnitrate-phosphate; tromantadine; Zole, efavirenz, eflornithine, eletriptan, emitricitabine, enflu tropatepine; tropenZiline-bromide; tropisetron hydrochlo rane, enoxacin, eprosartan, eptifibatide, ertapenem, escitalo ride; trospectomycin; troXerutin; troXipide; tulobuterol; pram, esmolol, esomeprazole, estazolam, estramustine, tybamate; tymazoline; tyrothricin, ubiquinone; udpg, ufe eSZopiclone, ethacrynate, ethacrynic acid, ethchlorVynol, namate; undecenoate calcium; unitiol; urapidil; uraZamide; ethiodized oil, ethionamide, ethopropazine, ethoXZolamide, valnoctamide: valproate-pivoxil, Valspodar, vasopressin; vel etidronate, etoposide, evans blue, eZetimibe, famotidine, nacrine; Veralipride; Vesnarinone; Vetrabutine; vigabatrin; felodipine, fenofibrate, fenoldopam, flecainide, floxuridine, Viloxazine-hydrochloride; viminol; vinburnine; Vincamine; fluconazole, flucytosine, fludarabine, fludeoxyglucose, Vincamine-teprosilate; Vindesine; Vinpocetine; vinylbital; fludrocortisone, flumazenil, flumethasone, flunisolide, fluo cinolone acetonide, fluocinonide, fluorometholone, fluo Vinzolidine Sulfate; Viprostol; vicquidil; virginiamycin; Visna rometholone, fluorouracil, fluoxetine, fluoxymesterone, dine: Xamoterol-fumarate; Xanoxic-acid; Xantofyl-palmitate; fluphenazine, fluiphenazine enanthate, fluiphenazine, flupred Xibornol; Xipamide; Xylazine; Zaprinast; Zimeldine; Zinosta nisolone, flurandrenolide, flurazepam, flurbiprofen, fluta tin, Zipeprol; Zolimidine; Zomepirac, Zorubicin; Zotepine; mide, fluticaSone, fluvastatin, fluvoxamine, fomivirsen, Zucapsaicin; and Zuclopenthixol-acetate. fondaparinux, foSamprenavir, foScarnet, fosfomycin 0082 Chemicals that are especially preferable to be used tromethamine, fosinopril, froVatriptan, fulvestrant, furo with the present invention include the following chemicals semide, gadobenate, gadodiamide, gadopentetate, gadoteri and their salts and free bases: abarelix, abciximab, acampro dol, gadoversetamide, gallium citrate, gallium salts, ganire sate, acetazolamide, acetohexamide, acetophenazine, acetri lix, gatifloxacin, gefitinib, gemcitabine, gemifloxacin, Zoate, acetylcysteine, adefovir dipivoxil, alatrofloxacin, glimepiride, glipizide, glucagon, glyburide, grepafloxacin, albendazole, alclometaSone, alendronate, almotriptan, alpra griseofulvin, guanabenz, guanfacine, halazepam, halobeta Zolam, ambenonium, amifostine, amiloride, amiodarone, Sol, halofantrine, haloperidol, haloprogin, halothane, hep amlodipine, amlodipine, amodiaquine, amoxapine, amoxicil arin, hexachlorophene, hydrochlorothiazide, hydrocortisone, lin, amplicillin, amprenavir, anagrelide, anakinra, apracloni hydroflumethiazide, hydroxocobalamin, hydroxychloro dine, aprepitant, ardeparin, argatroban, aripiprazole, artic quine, hydroxyzine, ibandronate, ibutilide, idoxuridine, ifos aine, atorvastatin, atovaquone, auranofin, azathioprine, famide, imipenem, indapamide, indium and its salts, indium aZelastine, azlocillin, aztreonam, bacampicillin, bacitracin, oxyquinoline, indium pentetate, indium, indocyanine green, baclofen, beclomethasone, bendroflumethiazide, benzthiaz indomethacin, iobenguane, iocetamic acid, iodamide, iodipa ide, betamethasone, bicalutamide, bismuth Subsalicylate, mide, iodixanol, iodohippurate, iofetamine, iohexol, iopami bleomycin, bosentan, bretylium, brimonidine, brinzolamide, dol, iopanoic acid, iopromide, iothalamate, iothalamate, US 2015/0309021 A1 Oct. 29, 2015 40 iotrolan, ioVersol, ioxaglate, ioXilan, ipodate, isoflurane, isof are especially preferable to use with the present invention lurophate, itraconazole, ketamine, ketoconazole, ketotifen, consist of the following list: Cytokines; chemokines; G pro lamivudine, lamotrigine, lanSoprazole, leflunomide, levami tein-coupled receptors; Glycosidases; glycosyltransferases; sole, levocabastine, levofloxacin, levothyroXine, lincomycin, Ion channels: Nuclear receptors: Phosphodiesterases; Pro lindane, lineZolid, liothyronine, lodoxamide, lomefloxacin, teases; Protein kinases; Protein phosphatases; Nucleic acids; lomustine, loperamide, loracarbef, loratadine, lorazepam, Transporter proteins; Serotonin receptors; Dopamine recep losartan, loteprednol, loxapine, mafenide, magnesium salts, tors; Adrenergic receptors; Histamine receptors; Muscarinic malathion, mangafodipir, manganese salts, mazindol, receptors; Nucleotide receptors; Cannabinoid receptors: mechlorethamine, meclizine, meclocycline, meclofenamate, Chemokine receptors; Opioid receptors; Tyrosine kinases; mefloquine, meloxicam, melphalan, mercaptopurine, mero Cytoplasmic serine kinases; cytoplasmic threonine kinases; penem, mersalyl, mesoridazine, methazolamide, methdila Thyroid hormone-like receptors; HNF3-like receptors; and Zine, methicillin, methimazole, methixene, methotrimepra Estrogen-like receptors. Proteins that are especially prefer Zine, methoxyflurane, methyclothiazide, metoclopramide, able to use with the present invention consist of the following metolaZone, metrizamide, mezlocillin, micafungin, micona list: 1.3.4.6-Tetrachloro-1,4-Cyclohexadiene H; 1,3,6,8-Tet Zole, midazolam, mitotane, modafinil, mometaSone, mon rahydroxynaphthalene Reductase; 1,3-1,4-Beta-Glucanase; telukast, moxalactam, moxifloxacin, nafcillin, naratriptan, 1,4-Alpha Maltotetrahydrolase; 1,4-Alpha-D-Glucan Glu nefazodone, nelfinavir, niclosamide, nilutamide, nitaZOX canohydrolase; 1,4-Beta-D-Glucan Cellobiohydrolase Cell: anide, nizatidine, nofetumomab, norfloxacin, octreotide, 1,4-Beta-D-Glucan Cellobiohydrolase I: 1,4-dihydropyri ofloxacin, olanzapine, omeprazole, oxacillin, oxaliplatin, dine Receptor on alpha1 subunit of L-type Voltage sensitive oxazepam, oxiconazole, oxytocin, pamidronate, pantopra Ca2+ channels; 10 Kda Chaperonin; 10-Formyltetrahydro Zole, paramethasone, paroxetine, pegaptainib, penicillamine, folate Dehydrogenase; 11-cis retinol dehydorgenase; penicilling benzathine, penicillin V, pentetate, pentosan 12-Oxophytodienoate Reductase; 12-Oxophytodienoate polysulfate, perflexane, perflubron, perfluoropolymethyliso Reductase 1: 12-Oxophytodienoate-10,11-Reductase: 14-3- propyl ether, perflutren, pergolide, permethrin, perphenazine, 3-Like Protein C: 15-hydroxyprostaglandin dehydrogenase phenoxybenzamine, pimecrolimus, pimozide, pioglitaZone, NAD+: 17-Beta-Hydroxysteroid Dehydrogenase; 17-Beta piperacetazine, piperacillin, pipobroman, piroxicam, polythi Hydroxysteroid Dehydrogenase 4; 17.KdFetal Brain Protein; azide, porfimer, potassium salts, povidone-iodine, pramipex 19-Mer Peptide Fragment Of Rhodopsin: 1-Aminocyclopro ole, pramlintide, prazepam, prednisolone, probenecid, probu pane-1-Carboxylate Deaminase: 1-Deoxy-D-Xylulose col, prochlorperazine, proguanil, promazine, promethazine, 5-Phosphate Reductois: 1-Pyrroline-5-Carboxylate Dehy propiomazine, propyliodone, propylthiouracil, drogenase; 1SY6:H OKT3 Heavy Chain 1; 1SY6:L OKT3 pyrimethamine, pyrithione Zinc, quazepam, quetiapine, Light Chain 1: 2,2-Dialkylglycine Decarboxylase: 2,3-Bis quinethaZone, quinupristin, rabeprazole, raloxifene, raniti phosphoglycerate-Independent Phosphog: 2,3-Dihydroxy dine, ranitidine bismuth citrate, riluzole, risedronate, risperi benzoate-Amp Ligase; 2.3-Dihydroxybiphenyl Dioxyge done, ritonavir, rofecoxib, rose bengal, rosiglitaZone, rosuv nase, 2,3-Dihydroxybiphenyl-1,2-Dioxygenase; 2,4- astatin, rubidium salts, Samarium lexidronam, Selenium Dienoyl-Coa Reductase; 2,5-Diketo-D-Gluconic Acid Sulfide; selenite salts, selenate salts, selenomethionine, Serta Reductase: 23-Kda Polypeptide Of Photosystem II Oxygen-: conazole, Sertraline, sevoflurane, Sibutramine, sildenafil, sil 23S Ribosomal; 23S Ribosomal RNA; 23S rRNA of 50S ver Sulfadiazine, simethicone-cellulose, sodium fluoride, ribosomal subunit; 25-hydroxyvitamin D-1 alpha hydroxy Sodium iodide, Sotalol, sparfloxacin, spirapril, spironolac lase, mitochondrial; 2'-5'-Oligoadenylate Synthetase 1; 26S tone, strontium chloride. Succimer, Sucralfate, Sufentanil, Sul proteasome non-ATPase regulatory subunit 1, 2-amino-3- conazole, Sulfacetamide, Sulfacytine, Sulfadiazine, Sulfadox ketobutyrate coenzyme Aligase: 2-Amino-4-Hydroxy-6-Hy ine, Sulfamerazine, Sulfameter, sulfamethizole, droxymethyldihydropte: 2-amino-4-hydroxy-6-hydroxym Sulfamethoxazole, Sulfanilamide, Sulfaphenazole, Sulfasala ethyldihydropteridine pyrophosphokinase; 2-Am Zine, Sulfinpyrazone, Sulfisoxazole, Sulfisoxazole acetyl, Sul inoethylphosphonate-Pyruvate Aminotr; 2-C-Methyl-D- foxone, Sulfur, Sulindac, Sumatriptan, Sumatriptan, Suprofen, Erythritol 2.4-Cyclodiphosphate: 2-C-Methyl-D-Erythritol tamsulosin, tazarotene, taZobactam, temazepam, teniposide, 4-Phosphate Cytidyly; 2C-Methyl-D-Erythritol-2,4-Cyclo tenofovir disoproxil, terconazole, thallous chloride, thia diphosphate; 2-Dehydro-3-Deoxyphosphooctonate Aldo bendazole, thiamine, thiamylal, thiethylperazine, thiogua lase: 2-Enoyl-Coa Hydratase: 2-Haloacid Dehalogenase; nine, thiopental, thioridazine, thiotepa, thiothixene, tiagab 2-Isopropylmalate Synthase: 2-Keto-3-Deoxy Gluconate ine, ticarcillin, ticlopidine, tiludronate, timolol, tinidazole, Aldolase: 2-Keto-3-Deoxygluconate Kinase, 2-oxoglutarate tinzaparin, tioconazole, tiopronin, tiotropium, tirofiban, tiza dehydrogenase E1 component, mitochondrial precursor; nidine, tolaZamide, tolbutamide, topiramate, toremifene, 2-Pyrone Synthase; 3 beta-hydroxysteroid dehydrogenase/ torsemide, travoprost, traZodone, triamcinolone, triamcino delta 5-; 3.2-Trans-Enoyl-Coa Isomerase, Mitochondrial; lone acetonide, triamcinolone diacetate, triamcinolone 3,4-Dihydroxy-2-Butanone 4-Phosphate Synthase; 3',5'-Cy hexacetonide, triazolam, trichlormethiazide, triclofos, tri clic Nucleotide Phosphodiesterase 2A; 30S ribosomal pro closan, trifluoperazine, triflupromazine, trifluridine, trime tein S12:30S ribosomal protein S4; 30S Ribosomal Protein prazine, trimethaphan, troglitaZone, trovafloxacin, tyro S6: 30S ribosomal protein S9; 32.1 Kda Protein. In Adh3 panoate, uracil mustard, Valdecoxib, valrubicin, Vancomycin, Rcal Intergenic; 33 Kda Chaperonin; 3'-5' Exonuclease Eri1; Vardenafil. Voriconazole, Zafirlukast, Ziconotide, Zileuton, 36 Kda Soluble Lytic Transglycosylase; 367Aa Long Hypo Ziprasidone, Zoledronic acid. The present invention is espe thetical Cytochrome P450; 385Aa Long Conserved Hypo cially useful when a solution of at least two chemicals is used, thetical Protein: 3-Alpha, 20-Beta-Hydroxysteroid Dehydro where the first chemical is selected from the above lists of genase; 3-Alpha-Hydroxysteroid Dehydrogenase; 3 Alpha chemicals and the second chemical does not have the same Hydroxysteroid Dehydrogenase Type 3: 3-Alpha empirical formula as the first chemical. Protein classes that Hydroxysteroid/Dihydrodiol Dehydrogease: 3-Carboxy-Cis, US 2015/0309021 A1 Oct. 29, 2015

Cis-Muconate Cycloisomerase; 3-Dehydroquinate Dehy drogenase; 7.8-Diamino-Pelargonic Acid Aminotransferase; dratase; 3-Dehydroquinate Dehydratase Arod; 3-Dehydro 7,8-Dihydro-6-Hydroxymethylpterin-Pyrophosp; 70 Kilo quinate Synthase; 3-Deoxy-D-Arabino-Heptulosonate-7- dalton Heat Shock Protein: 70-Kda Soluble Lytic Transgly Phosphatase: 3-Deoxy-Manno-Octulosonate cosylase; 7-dehydrocholesterol reductase; 8-Amino-7-Ox Cytidylyltransfer; 3-Hydroxy-3-Methylglutaryl-Coa Syn ononanoate Synthase; 8-Oxoguanine DNA Glycosylase; 92 thase;3-Hydroxy-3-Methylglutaryl-Coenzyme A Reductase: Kda Type IV Collagenase; A chain; A/G-Specific Adenine 3-Hydroxyacyl-Coa Dehydrogenase; 3-Hydroxyacyl-Coa Glycosylase; Aac, Aah2: Lah-Alpha-It. Abc Transporter, Dehydrogenase Type II: 3-hydroxyisobutyrate dehydroge ATP Binding Protein: Acetate Kinase; Acetoacetyl-Coa Thio nase, mitochondrial precursor, 3-Isopropylmalate Dehydro lase; Acetohydroxy-Acid lSomeroreductase; Acetohydroxy genase; 3-Ketoacetyl-Coa Thiolase: 3-Keto-L-Gulonate Acid Synthase; Acetoin Reductase; Acetolactate Synthase, 6-Phosphate Decarboxylase; 3-keto-steroid reductase: Catabolic; Acetolactate Synthase, Mitochondrial; Acetyl 3-Mercaptopyruvate Sulfurtransferase: 3-Methyl-2-Oxobu Group: Acetyl Transferase: Acetyl Xylan Esterase: Acetyl tanoate Hydroxymethyltra; 3-Methyl-2-Oxobutanoate choline Binding Protein; Acetylcholine-Binding Protein; Hydroxymethyltransfe: 3-Methyladenine DNA Glycosylase; Acetylcholinesterase: Acetylcholinesterase; Acetylcho 3-Methylaspartate Ammonia-Lyase: 3-oxo-5-alpha-steroid linesterase (AChE); Acetylcholinesterase precursor; Acetyl 4-dehydrogenase 2: 3-Oxoacyl-, 3-Oxoacyl-Acyl-Carrier Coa Acetyltransferase; Acetyl-CoA Acetyltransferase (Mito Protein Reductase; 3-Oxoacyl-Acyl-Carrier Protein Syn chondrial); Acetyl-Coa Carboxylase; Acetyl-CoA thas; 3-Oxoacyl-Acyl-Carrier-Protein Synthas; 3-oxoacyl carboxylase 2: Acetyl-Coa Synthetase; Acetyl-Coenzyme A acyl-carrier-protein synthase I, 3-oxoacyl-acyl-carrier Acetyltransferase 2: Acetyl-Coenzyme A Carboxylase; protein synthase II: 3-oxoacyl-acyl-carrier-protein Acetyl-Coenzyme A Synthetase 1: Acetyl-coenzyme A Syn synthase III: 3-Phosphoglycerate Kinase; 3-Phosphoi thetase 2-like, mitochondrial; Acetyl-coenzyme A Syn nositide Dependent Protein Kin: 3-Phosphoinositide Depen thetase, cytoplasmic; Acetylglutamate Kinase; Acidic Fibro dent Protein Kinase-, 3-Phosphoshikimate 1-Carboxyvinyl blast Growth Factor; Acidic Lectin; Acireductone transferase: 3-Phytase A; 47 Kda Membrane Antigen; Dioxygenase; Aclacinomycin Methylesterase; Aclacinomy 4-Alpha-Glucanotransferase: 4-Amino-4-Deoxychorismate cin-10-Hydroxylase; Aconitase; Aconitate Hydratase; Aconi Lyase, 4-Aminobutyrate Aminotransferase: 4-Chloroben tate Hydratase 2: Acriflavine Resistance Protein B; Actagar Zoyl Coa Ligase, 4-Chlorobenzoyl Coenzyme A Dehaloge dine; Actin; Actin Alpha 1: Actin Depolymerizing Factor; nase: 4-Cresol Dehydrogenase Hydroxylating Flavo; Actin, Alpha Skeletal Muscle; Actin-Fragmin Kinase; Acti 4-Diphosphocytidyl-2-C-Methyl-D-Erythrit: 4-Diphospho vated Cdc42 Kinase 1: Activator Of Activin receptor; Activin cytidyl-2-C-Methylerythritol Synt: 4-Hydroxybenzoyl-Coa Receptor Type II: Actva-Orf6 Monooxygenase; Acutohae Thioesterase: 4-Hydroxyphenylpyruvate Dioxygenase; monlysin; Acyl Carrier. Protein; Acyl Carrier Protein Phos 4-Hydroxyphenylpyruvic Acid Dioxygenase: 4-Hydroxyth phodiesterase; Acyl Carrier Protein Synthase; Acyl carrier reonine-4-Phosphate Dehydrogenase; 4M5.3 Anti-Fluores protein, mitochondrial precursor; Acyl-Acyl-Carrier Pro cein Single Chain Antibody: 4-Oxalocrotonate Tautomerase: tein Desaturase; Acyl-Acyl-Carrier-Protein-Udp-N- 4'-Phosphopantetheinyl Transferase Sfp: 4-trimethylami Acetylgl; Acylamino-Acid-Releasing Enzyme; Acyl-Coa nobutyraldehyde dehydrogenase; 5,10-Methenyltetrahydro Dehydrogenase; Acyl-CoA dehydrogenase family member 8; folate Synthetase; 5,10-Methylenetetrahydrofolate Dehydro Acyl-Coa Dehydrogenase Family Member 8, Mito. Acyl-Coa genase; 5,10-Methylenetetrahydrofolate Reductase; 50S Dehydrogenase, Medium-Chain Specifi; Acyl-CoA dehydro Ribosomal Protein LIP; 50S subunit of the 70S ribosome of genase, short-chain specific, mitochondrial precursor, Acyl bacteria: 5-alpha reductase 1: 5-Aminolaevulinic Acid Dehy Coa Hydrolase; Acyl-Coa Oxidase; Acyl-Coa Thioesterase I; dratase; 5-aminolevulinate synthase; 5-Aminolevulinic Acid Acyl-Coa Thioesterase II: Acyl-Coenzyme A Binding Pro Dehydratase; 5'-AMP-activated protein kinase, beta-2 sub tein; acyl-coenzyme A:cholesterol acyltransferase; unit; 5'-AMP-activated protein kinase, catalytic alpha-1 Acylphosphatase; Adam 33, Adamalysin II: Adenine Glyco chain; 5'-D: 5'-Deoxy-5'-Methylthioadenosine Phosphory sylase; Adenine Phosphoribosyltransferase: Adenine-N6 las; 5-Enolpyruvylshikimate-3-Phosphate Synthase: 5-Epi DNA-Methyltransferase Taqi; Adenosine 2B receptor; Aristolochene Synthase; 5'-Fluoro-5'-Deoxyadenosine Syn Adenosine A1 receptor, Adenosine A3 receptor, Adenosine thase; 5-HT-1A Receptor; 5-HT-1 B Receptor; 5-HT-1 D Deaminase; Adenosine Kinase; Adenosylcobinamide Receptor; 5-HT2A Receptor; 5-HT3 Receptor; 5HT4 recep Kinase; Adenosylhomocysteinase; Adenosylmethionine-8- tor; 5-hydroxytryptamine 1E receptor; 5-hydroxytryptamine Amino-7-Oxononanoat, Adenosylmethionine-8-Amino-7- 1F receptor; 5-hydroxytryptamine 2C receptor; 5-hydrox Oxononanoate Am; Adenylate Cyclase; Adenylate cyclase ytryptamine 7 receptor; 5-Methyltetrahydrofolate S-Ho (cAMP); Adenylate Kinase; Adenylate Kinase 1: Adenylate mocysteine; 5-Methyltetrahydrofolate S-Homocysteine Kinase Isoenzyme-3, AdenyloSuccinate Synthetase; Adeny Meth: 5-Methyltetrahydropteroyltriglutamate-, 5-Methyltet losuccinate synthetase isozyme 1: Adenylyl Cyclase; Adeny rahydropteroyltriglutamate-Homo; 5'-Methylthioadenosine lylsulfate Kinase; Adipocyte Lipid Binding Protein; Adipo Phosphorylase; 5'-Nucleotidase; 5'-R: 6,7-Dimethyl-8-Ribi cyte Lipid-Binding Protein; ADP Compounds Hydrolase tyllumazine Synthase; 6-5 Deoxyerythronolide B Hydroxy Nude; ADP, ATP Carrier Protein Heart Isoform T1:ADP ATP lase; 6-Hydroxymethyl-7,8-Dihydropterin Pyrophosph; carrier protein, fibroblast isoform; ADP, ATP carrier protein, 6-Oxocamphor Hydrolase; 6-Phospho-Beta-D-Galactosi heart/skeletal muscle isoform T1: ADP, ATP carrier protein, dase; 6-Phospho-Beta-Glucosidase 6-Phosphofructo-2-Ki liver isoform T2; ADP-Dependent Glucokinase; ADP-L- nase/Fructose-2,6-Bisp; 6-Phosphofructo-2-Kinase/Fruc Glycero-D-Mannoheptose 6-Epimerase: Adpr Pyrophos tose-2,6-Bipho; 6-Phosphofructo-2-Kinase/Fructose-2,6- phatase; ADP-Ribose Pyrophosphatase; ADP-Ribosyl Bisph; 6-Phosphofructokinase; 6-Phosphogluconate Cyclase; ADP-Ribosylation Factor 1; ADP-Ribosylation Dehydrogenase; 6-Phosphogluconolactonase; 6-Pyruvoyl Factor 2; ADP-Ribosylation Factor 4: ADP-Ribosylation Tetrahydropterin Synthase; 7 Alpha-Hydroxysteroid Dehy Factor 6: ADP-Ribosylation Factor-Like 8: ADP-Ribosyla US 2015/0309021 A1 Oct. 29, 2015 42 tion Factor-Like Protein 1; ADP-Ribosylation Factor-Like charin Complexed With Adenine: Alpha-Momorcharin Com Protein 3; ADP-Ribosylation Factor-Like Protein 5; ADP plexed With Formycin; Alpha-N-Acetylgalactosaminidase; Ribosyltransferase; Adrenodoxin; Adrenodoxin Reductase: Alpha-N-acetylgalactosaminidase precursor, Alpha-N- Adsorption Protein P2: Aequorin; Aequorin 2: Aeromonas acetylglucosaminidase precursor; Alpha-Neurotoxin Tx12; caviae: AFG3-like protein 1: AFG3-like protein 2: Aggluti Alpha-Tocopherol Transfer Protein; Alpha-Trehalose-Phos nin; Agglutinin; Agglutinin Isolectin 3: Agglutinin Isolectin phate Synthase; Alpha-Trichosanthin Complexed With I/Agglutinin Isolect, Agglutinin Isolectin Vi; Agglutinin Adenine; Amic; Amicyanin; Amidase Operon; Amidophos Isolectin Vi/Agglutinin Isolec.; Agmatinase; Agmatine Imino phoribosyltransferase: Amidophosphoribosyltransferase hydrolase; Agrobacterium Tumefaciens Dps; Ahplaao; Aicar precursor; Amidophosphoribosyltransferase precursor; Transformylase-Imp Cyclohydrolase; Alamethicin; Alanine Amidotransferase Hish; Amiloride-sensitive amine oxidase aminotransferase; Alanine Dehydrogenase; Alanine Race copper-containing precursor; Amiloride-sensitive cation mase; Alanine racemase, biosynthetic; Alanine-Glyoxylate channel 1, neuronal; Amiloride-sensitive cation channel 2, Aminotransferase; Alanine-glyoxylate aminotransferase 2; neuronal; Amiloride-sensitive Sodium channel alpha-Subunit; Alanyl-tRNA synthetase; Alcohol Dehydrogenase; Alcohol Amiloride-sensitive sodium channel beta-subunit; Dehydrogenase; Alcohol dehydrogenase NADP+: Alcohol Amiloride-sensitive Sodium channel delta-subunit; dehydrogenase 6: Alcohol Dehydrogenase Alpha Chain; Amiloride-sensitive sodium channel gamma-Subunit; Amine Alcohol Dehydrogenase Beta Chain; Alcoholdehydrogenase Oxidase; Amine Oxidase Flavin-Containing A; Amine Oxi class II pi chain precursor; Alcohol Dehydrogenase Class III dase Flavin-Containing. B; Amine Oxidase, Copper Con Chi Chain; Alcohol dehydrogenase class IV mu?sigma chain; taining; Amine Oxidase, Flavin-Containing; Aminoacylase Alcohol Dehydrogenase Class IV Sigma Chain; Alcohol 1; Aminoglycoside 2'-N-acetyltransferase; Aminoglycoside Dehydrogenase E Chain; Alcohol Dehydrogenase Gamma 3'-Phosphotransferase; Aminoglycoside 6'-N-Acetyltrans Chain; Alcohol Dehydrogenase, Class II; Alcohol Dehydro ferase; Aminomethyltransferase; Aminopeptidase; Ami genase, Class II; Alcohol Dehydrogenase, Iron-Containing; nopeptidase P, Aminotransferase; Aminotransferase, Puta Alcohol Sulfotransferase; Aldehyde Dehydrogenase; Alde tive; Amnionless protein; AMP deaminase 1: Amp hyde Dehydrogenase; Aldehyde dehydrogenase 1A3; Alde Nucleosidase; AMP-activated protein kinase (AMPK); AMT hyde dehydrogenase 3B1; Aldehyde dehydrogenase 3B2: protein; Amyloid Beta-Peptide; Amyloid protein-binding Aldehyde dehydrogenase family 7 member A1: Aldehyde protein 1; Amylomaltase; Amylosucrase; Anabaena Sensory dehydrogenase X, mitochondrial precursor; Aldehyde Dehy Rhodopsin; Anaerobic Ribonucleotide-Triphosphate Reduct; drogenase, Cytosolic 1: Aldehyde dehydrogenase, dimeric Androgen Receptor; Angiogenin; Angiostatin; Angiotensin NADP-preferring; Aldehyde Dehydrogenase, Mitochondrial Converting Enzyme; Angiotensin-converting enzyme (ACE); Precur; Aldehyde dehydrogenase, mitochondrial precursor; Annexin A1, Annexin III; Annexin V: Anthocyanidin Syn Aldehyde Ferredoxin Oxidoreductase Protein C: Aldehyde thase; Anthranilate Phosphoribosyltransferase; Anthranilate oxidase; Aldehyde Oxidoreductase; Aldehyde Reductase: Phosphoribosyltransferase 2: Anthrax Toxin Receptor 2: Anti Aldo-Keto Reductase Family 1 Member C1; Aldo-keto IgE antibody VH domain chain 1: Anti IgE antibody VL reductase family 1 member C2; Aldo-Keto Reductase Family domain chain 1; Antibody Vhh Lama Domain; Antifungal 1 Member C3; Aldo-keto reductase family 1 member C4; Protein 1: Antigen 85B; Antigen 85-C; Anti-H; Anti-Muelle Aldolase; Aldolase Protein; Aldose 1-Epimerase; Aldose rian hormone type II receptor; Anti-Platelet Protein; Anti Reductase; Alginate Lyase; Algal; Algo2; ALK tyrosine Sigma F Factor Antagonist; Antithrombin III (ATM); Anti kinase receptor; Alkaline Phosphatase: Alkaline Phos thrombin-III; Antiviral Protein 3; Antiviral Protein S. Apag phatase: Alkyl Hydroperoxide Reductase Subunit F: Allan Protein; Apc35852; Apc35880; Apical Membrane Antigen 1: toate Amidohydrolase; Allene Oxide Synthase-Lipoxyge Apocarotenoid-Cleaving Oxygenase; Apoferritin Co-Crys nase Protein; Alliin Lyase; Alpha Amylase; Alpha tallized With Sn-Protopor; Apollipoprotein: Apolipoprotein Glutathione S-Transferase: Alpha, Alpha-Trehalose-Phos A-II; Apoptosis regulator Bcl-2: Apoptotic Protease Activat phate Synthase; Alpha-1 Catenin; Alpha1.2-Mannosidase; ing Factor 1; Apyrase; Aquaporin 1: Aquaporin Z: Arabinan Alpha-1,2-Mannosidase; Alpha-1,3-Mannosyl-Glycoprotein Endo-1,5-Alpha-L-Arabinosidase A: Arabinan-Endo 1.5-Al Beta-1,2-N : Alpha-1,4-Galactosyl Transferase: Alpha-1,4- pha-L-Arabinase; Arabinose Operon Regulatory Protein; Glucan-4-Glucanohydrolase; Alpha-1,4-N-Acetylhexosami Arac, Arachidonate 15-lipoxygenase; Arachidonate 5-li nyltransferase Ext; Alpha-1A Adrenergic Receptor; Alpha1 poxygenase; A-Rafproto-oncogene serine/threonine-protein Antitrypsin; Alpha-1-Antitrypsin, Alpha-1B adrenergic kinase; Arcelin-1; Arcelin-5A; Archaeal Sm-Like Protein Af receptor; Alpha-1D adrenergic receptor; Alpha-1-Purothio Sm2; Archaeosine tRNA-Guanine Transglycosylase; Argin nin; Alpha-2.3/8-Sialyltransferase: Alpha-2A Adrenergic ase; Arginase 1: Arginase I; Arginase II; Arginase II, Mito Receptor; Alpha-2B adrenergic receptor; Alpha-2C adrener chondrial Precursor, arginine decarboxylase; Arginine gic receptor; Alpha-2-Macroglobulin; Alpha-2U-Globulin; Kinase; Arginine N-Succinyltransferase, Alpha Ch; Argini Alpha-aminoadipic semialdehyde synthase, mitochondrial noSuccinate Synthase; Argininosuccinate synthase Frag precursor, Alpha-Amylase; Alpha-Amylase I: Alpha-Amy ment; Argininosuccinate Synthetase; Arginosuccinate lyase; lase II: Alpha-Amylase Isozyme 1: Alpha-Amylase, Pancre Arginosuccinate synthase; Aristolochene Synthase; Arnb atic; Alpha-Amylase, Salivary; AlphaChain (LH); Alpha Aminotransferase; Arno; Aromatase; Aromatic Amino Acid Conotoxin Gid: Alpha-D-Glucose 1,6-Bisphosphate Aminotransferase; Arpg836; Arsenate Reductase; Arsenical Phosphotran; Alpha-D-Glucuronidase; Alpha-Galactosi pump-driving ATPase; Arsenical Resistance Operon Repres dase; Alpha-Galactosidase A, Alpha-Glucosidase; Alpha Sor, Pu; Arsenite-Translocating Atpase; Arthropodan Glucosidase; Alpha-Glucuronidase; Alpha-Ketoglutarate Hemocyanin; Artificial Nucleotide Binding Protein; Arto Dependent Taurine Dioxyg: Alpha-L-Arabinofuranosidase; carpin; Aryl Sulfotransferase; Arylamine N-Acetyltrans Alpha-L-Arabinofuranosidase B: Alpha-Lytic Protease: ferase; Arylsulfatase; Arylsulfatase A: Arylsulfatase A: Alpha-Mannosidase II: Alpha-Momorcharin: Alpha-Momor Ascorbate Oxidase; Ascorbate Peroxidase; Asparagine Syn US 2015/0309021 A1 Oct. 29, 2015

thetase; Asparagine Synthetase B; Asparaginyl-tRNA syn thetase; Beta-Lactamase; Beta-Lactamase CtX-M-14; Beta thetase; Aspartate 1-Decarboxylase; Aspartate 1-Decarboxy Lactamase CtX-M-27; Beta-Lactamase CtX-M-9; Beta-Lac lase Precursor, Aspartate Aminotransferase; Aspartate tamase II: Beta-Lactamase Imp-1: Beta-Lactamase Oxa-1: Aminotransferase; Aspartate aminotransferase, cytoplasmic; Beta-Lactamase Oxa-10; Beta-Lactamase Oxa-2: Beta-Lac Aspartate aminotransferase, mitochondrial; Aspartate Dehy tamase Pse-2. Beta-Lactamase Shy-1: Beta-Lactamase Shv drogenase; Aspartate Receptor, Aspartate Transcarbamoy 2. Beta-Lactamase Tem; Beta-Lactamase. Type II: Beta-Lac lase; Aspartate-Semialdehyde Dehydrogenase; Aspartic Pro toglobulin; Beta-Mannanase; Beta-Mannosidase; Beta tease Bla G. 2; Aspartic Proteinase; Aspartoacylase; Asparty1 Methylaspartase: Beta-Momorcharin; Beta-N- aminopeptidase; Asparty1/asparaginyl beta-hydroxylase; Acetylhexosaminidase; Beta-N-Acetylhexosaminidase; Aspartyl-tRNA Synthetase: Aspergillopepsin; Assemblin; Beta-Phosphoglucomutase; Beta-Purothionin; Beta-Secre Astacin: At5G 11950; ATP Phosphoribosyltransferase: ATP tase 1: Beta-Spectrin; Beta-Trypsin; Beta-Tryptase: Bh()236 Sulfurylase: ATP Synthase; ATP synthase delta chain, mito Protein; Bifunctional 3'-Phosphoadenosine 5'-Phospho: chondrial Precursor; ATP-binding cassette: ATP-Depen Bifunctional adenosylcobalamin biosynthesis protein cobU: dent Clp Protease ATP-Binding Subun; ATP-Dependent Bifunctional aminoacyl-tRNA synthetase: Bifunctional DNA Helicase: ATP-Dependent Hsl Protease ATP-Binding Deaminase/Diphosphatase: Bifunctional Dihydrofolate S; ATP-Dependent Metalloprotease Ftsh; ATP-Dependent Reductase-Thymidy; Bifunctional dihydrofolate reductase RNA Helicase P54: ATP-Phosphoribosyltransferase: ATP thymidylate synthase: Bifunctional Histidine Biosynthesis sensitive inward rectifier potassium channel 1: ATP-sensitive Prot; Bifunctional methylenetetrahydrofolate dehydroge inward rectifier potassium channel 11; Atrial Natriuretic Pep nase/cyclohydrolase; Bifunctional P-450: Nadph-P450 tide Clearance Recepto; Atrial natriuretic peptide receptor A: Reductase: Bifunctional PGK/TIM Includes: Phosphoglyc Atrial Natriuretic Peptide Receptor A; Atrolysin C; Aug erate kinase, EC 2.7.2.3, Triosephosphate isomerase, EC5.3. menter Of Liver Regeneration: Aurora-Related Kinase 1: 1.1, TIM, Triose-phosphate isomerase: Bifunctional Purine Autocrine Motility Factor; Autoinducer-2 Production Protein Biosynthesis Protein Pur: Bifunctional purine biosynthesis Luxs; Autolysin; Auxin Binding Protein 1: Avermectin-Sen protein PURH Includes: Phosphoribosylaminoimidazole sitive Chloride Channel GI; Avian Sarcoma Virus Integrase: carboxamide formyltransferase, EC 2.1.2.3, AICAR trans Avidin; Axin; AZurin; B chain; B Lymphocyte Stimulator; formylase, IMP cyclohydrolase, EC 3.5.4.10, Inosinicase, B4Dimer; B9340; Bacterial azoreductase (Bacillus sp); Bac IMP synthetase, ATIC: Bifunctional Puta Protein: Bifunc terial isoleucyl-tRNA synthetase; Bacterial Leucyl Ami tional Rela/Spot; Bikunin; Bile Acid Receptor; Bile salt nopeptidase; Bacterial membranes; Bacterial Sialidase; Bac export pump; Bile-Salt Activated Lipase; Biliary Glycopro terial Sulfite Oxidase; Bactericidal/Permeability-Increasing tein C: Bilin Binding Protein; Bilin-Binding Protein; Biliver Protein; Bacteriochlorophyll A Protein; Bacterioferritin: din IX Beta Reductase; Biliverdin Reductase A; Biliverdin Bacteriophage Lambda Lysozyme; Bacteriophage T4 Short reductase A precursor: Bioh Protein; Biosynthetic Thiolase; Tail Fibre; Bacteriorhodopsin; Baculoviral lap Repeat-Con Biotin Synthase; biotinidase; biotin-protein ligase; Biphenyl taining Protein 4; Bap1; Basement Membrane Protein 2,3-Diol 1,2-Dioxygenase; Bleomycin Resistance Determi Bm-40; Basic Fibroblast Growth Factor; Basic Phospholi nant; Bleomycin Resistance Protein; Bleomycin-Binding pase A2: Bba1; Bba5: B-cell receptor; Benzoate 1,2-Dioxy Protein; Blood Coagulation Factor Vii; Blue Fluorescent Pro genase Reductase; BenZodiazepine Receptor, Benzoylfor tein; B-Luffin; Bm-40; Bone Marrow Stromal Cell Antigen 1: mate Decarboxylase; Benzyl Alcohol Dehydrogenase; Beta 1 Bone Morphogenetic Protein 2: Bone Morphogenetic Protein adrenergic receptor, Beta 1.4 Galactosyltransferase; Beta 2 7; Botulinum Neurotoxin Type B; Bovine Beta-Lactoglobu adrenergic receptor; beta chain (FSH); Beta Crystallin B1; lin A; Bowman-Birk Inhibitor Derived Peptide; Bp40; Brady Beta Lactamase; Beta platelet-derived growth factor receptor kinin; B-Raf proto-oncogene serine/threonine-protein precursor, Beta Trypsin; Beta-(1,3)-glucan synthase Frag kinase; Branched-chain alpha-ketoacid dehydrogenase ment: Beta-1,4-D-Glycanase Cex-Cd; Beta-1,4-Galacta kinase; Branched-Chain Amino Acid Aminotransferase; nase; Beta-1,4-Galactosyltransferase, Beta-1,4-Galactosyl Branched-Chain Amino Acid Aminotransferase; Branched transferase 1: Beta-1,4-Mannanase; Beta2-Glycoprotein-1; chain-amino-acid aminotransferase, cytosolic; Branched Beta3 Alcohol Dehydrogenase; Beta-adrenergic receptor chain-amino-acid aminotransferase, mitochondrial; kinase 1: Beta-adrenergic receptor kinase 2: Beta-Agarase A: Brazzein; Bruton S Tyrosine Kinase; Bseó341 Restriction Beta-Alanine Synthase: Beta-Amylase; Beta-B2-Crystallin; Endonuclease; Butyryl-Coa Dehydrogenase; C Protein Beta-Carbonic Anhydrase: Beta-Catenin; BetaChain; Alpha-Antigen; C. pasteurianum Hydrogenase I: C-1027 BetaChain (LH); Beta-Conglycinin, Beta Chain; Beta-Cryp Apoprotein; C1R Complement Serine Protease: C-1-tetrahy togein; Beta-D-Glucan Exohydrolase Isoenzyme Exo 1: drofolate synthase: C-4 methylsterol oxidase; C4-Dicarboxy Beta-D-Glucan Glucohydrolase Isoenzyme Exo1; Beta-Elic late Transport Transcriptional R; Caffeic Acid 3-O-Methyl itin Cryptogein; Beta-Fructosidase; Beta-Galactosidase; transferase; Caffeoyl-Coa O-Methyltransferase: Cag-Z: Beta-Galactoside-Binding Lectin; Beta-Glucosidase; Beta Calcineurin B subunit isoform 1: Calcineurin B-Like Protein Glucosidase A. Beta-Glucosyltransferase: Beta-Glucu 4, Calcitonin, Calcitonin Analogue; Calcitonin receptor, Cal ronidase; Beta-Hexosaminidase; Beta-Hexosaminidase Beta cium/Calmodulin-Dependent 3',5'-Cycli; Calcium/Calm Chain; Beta-Hordothionin; Beta-Hydroxydecanoyl Thiol odulin-Dependent Protein Kinase; calcium-activated potas Ester Dehydrase; Betaine-Homocysteine Methyltransferase: sium channel; Calcium-binding mitochondrial carrier protein Betaine-Homocysteine S-Methyltransferase; Beta-Keto Acyl Aralarl; Calcium-binding mitochondrial carrier protein Carrier Protein Reductase; Beta-Ketoacyl Acyl Carrier Pro Aralar2: Calcium-Dependent Protease, Small Subunit; Cal tein Synthas; Beta-Ketoacyl-Acyl-Carrier-Protein Syn cium-transporting ATPase type 2C member 1, CalgizZarin; thas; Beta-Ketoacyl-Acp Synthase III: Beta-Ketoacyl-Acyl Calmodulin, Calpain 1, Large Catalytic Subunit; Camp Carrier Protein Synth; Beta-Ketoacyl-Acyl Carrier Protein Dependent Protein Kinase; cAMP Phosphodiesterase: Synthase; Beta-Ketoacylsynthase III: Beta-Lactam Syn Camp-Dependent Protein Kinase; Camp-Dependent Protein US 2015/0309021 A1 Oct. 29, 2015 44

Kinase Regulatory Sub: Camp-Dependent Protein Kinase Chalcone-Flavonone Isomerase 1: Chaperone Protein Htpg: Type 1; Camp-Dependent Protein Kinase Type I-Alpha R: C-Ha-Ras; Charybdotoxin; Chemosensory Protein A6; Camp-Dependent Protein Kinase, Alpha-Cat; Camp-Specific Chemotactic Peptide; Chemotaxis Protein Chea; Chemotaxis 3',5'-Cyclic Phosphodieste; Camp-Specific 3',5'-Cyclic Phos Protein Chey; Chemotaxis Receptor Methyltransferase Cher; phodiesterase; cAMP-specific 3',5'-cyclic phosphodiesterase Chimera Of Glutathione S-Transferase-Synthet; Chimera Of 7B; Camp-Specific Phosphodiesterase Pde4D2: Cannabinoid Maltose-Binding Periplasmic Prote: Chitinase; Chitinase; receptor 2; Cannabinoid Receptors; Capsid Protein C; Capsid Chitinase 1: Chitinase A; Chitinase A; Chitinase B: Chitinase Protein P40; Capsid Protein P40: Assemblin Protease; Cara: B: Chitinase-3 Like Protein 1; Chitobiase; Chitobiose Phos Carbapenem Synthase; Carbohydrate-Phosphate Isomerase: phorylase; Chitosanase; Chitotriosidase; Chitotriosidase 1: Carbon Monoxide Oxidation System Transcripti; Carbonic Chitotriosidase Synonym: Chitinase; Chloramphenicol Anhydrase; Carbonic Anhydrase I; Carbonic Anhydrase II; Acetyltransferase; Chloramphenicol Phosphotransferase: Carbonic Anhydrase III; Carbonic Anhydrase IV: Carbonic Chlorella Virus DNA Ligase-Adenylate; Chloride channel Anhydrase Xii; Carbonic Anhydrase Xiv; Carbonyl Reduc protein 2: Chloride Intracellular Channel Protein 1: Chloro tase; Carbonyl Reductase Nadph 1; Carboxy Methyl Trans catechol 1,2-Dioxygenase; Chloroperoxidase; Chloroperoxi ferase For Protein Phosp; Carboxy-Cis, Cis-Muconate dase F. Chlorophyll A-B Binding Protein Ab80; Chlorophyll Cyclase; Carboxyethylarginine Synthase; Carboxylesterase: A-B Binding Protein, Chloroplast; Chloroplast Ferredoxin Carboxylesterase Est2: Carboxylesterase Precursor; Car Nadp+Oxidoreductase; Chloroplast Outer Envelope Protein boxymethylated Rhodanese: Carboxymuconolactone Decar Oep34; Chloroplastic Ascorbate Peroxidase; Cho Reductase: boxylase Family Pr; Carboxypeptidase A; Carboxypeptidase Cholecystokinin type A receptor; Cholera Toxin: Cholera Gp180 Residues 503-882: Carboxypeptidase M: Carboxy Toxin B Subunit; Cholesterol Esterase; Cholesterol Oxidase; Terminal Domain RNA Polymerase I; Carminomycin 4-O- Choline dehydrogenase; Choline kinase alpha; Choline Methyltransferase; Carnitine Acetyltransferase; Carnitine acetyltransferase; Choline/ethanolamine kinase; Choline Acetyltransferase Isoform 2; Carnitine O-acetyltransferase: phosphate cytidylyltransferase A. Choline-phosphate cytidy Carnitine O-palmitoyltransferase I, mitochondrial liver iso lyltransferase B: Cholinesterase; Choloylglycine Hydrolase; form (CPT-1); Carnitine O-palmitoyltransferase II, mito Chondroitin Ac Lyase; Chondroitinase Ac; Chondroitinase chondrial (CPT-2); Casein Kinase II, Alpha Chain; Casein B: Chorionic Gonadotropin: Chorismate Lyase; Chorismate Kinase-1, Caspase-1 precursor; Caspase-3; Caspase-7: Cata Mutase; Chorismate Synthase; Chromosomal Replication bolic Alanine Racemase Dadx; Catabolite Gene Activator; Initiator Protei: Chymase; Chymase; Chymotrypsin; chymot Catabolite Gene Activator Protein: Catalase; Catalase 1: rypsin B: Chymotrypsinogen A; Circadian Clock Protein Catalase A: Catalase Hpii; Catalase-Peroxidase; Catalase Kaib; Cis-Biphenyl-2,3-Dihydrodiol-2,3-Dehydrogena; Peroxidase Protein Katg; Catechol 1,2-Dioxygenase; Cat Cite: Citrate Lyase, Beta Subunit; Citrate Synthase: Citrate echol Oxidase; Catechol-O-Methyltransferase: Catechol-O- Synthase: C-Kit Protein; Claret Segregational Protein; Class methyltransferase (COMT): Cathepsin B: Cathepsin F; BAcid Phosphatase; Class B Carbapenemase Blab-1; Class C Cathepsin G: Cathepsin K; Cathepsin S: Cathepsin V: Cation Beta-Lactamase; Class I Alpha-1,2-Mannosidase; Class-Mu Dependent Mannose-6-Phosphate Recepto: Cation-Depen Glutathione S-Transferase; Clavaminate Synthase 1: Clpb dent Mannose-6-Phosphate Recepto: Cationic amino acid Protein; Cmp-N-Acetylneuraminic Acid Synthetase; Coagul transporter-3; Cationic amino acid transporter-4; Cation-In lation Factor Ix; Coagulation Factor V: Coagulation factor dependent Mannose 6-Phosphate Recep: Cbp21; Cd2; CD2 VII: Coagulation Factor VIII; Coagulation Factor Viii Precur lymphocyte antigen; CD20 B-lymphocyte antigen; CD33 sor; Coagulation Factor X: Coagulation Factor Xiii; Coagul antigen; Cd46; CD52 Antigen; Cd58; Cd59 Complexed With lation Factor XiiiA Chain; Coagulation factor XIII A chain Glcnac-Beta-1,4-Glcnac-Be; Cdc25 B-Type Tyrosine Phos precursor; Coat Protein; Coatomer Gamma Subunit; Cob: phatase; Cdc42Hs-GDP; Cdp-Glucose 4,6-Dehydratase: Cob(I)yrinic acid a,c-diamide adenosyltransferase; Cobal Cdp-Glucose-4,6-Dehydratase; Cdp-Tyvelose-2-Epimerase: amin-Dependent Methionine Synthase; Cobalt-Precorrin-4 CEA or carcinoembryonic antigen-multiple sequences; Cell Transmethylase; Cocaine Esterase: Coenzyme A Biosynthe Cycle Arrest Protein Bub3; Cell Division Control Protein 2 sis Bifunctional Protein: Coenzyme F420-Dependent N5, Homolog: Cell Division Control Protein 6: Cell Division N10-Methylenete: Coenzyme F420H2: Nadp+ Oxidoreduc Inhibitor; Cell Division Protein Ftsy: Cell division protein tase; Coenzyme Paq Synthesis Protein C: Cog4826: Serine ftsZ: Cell Division Protein FtsZ. Homolog 1: Cell Division Protease Inhibitor; Collagen; Collagenase 3: Collagenase-3: Protein Kinase 2: Cell Division Protein Kinase 7: Cell Divi Collagen-Like Peptide; Complement C1R Component; sion Protein Zipa: Cell Wall Pentapeptide Ala-: Cell Wall Complement C1S Component; Complement C3Dg; Comple Peptide In Complex With Deglucobalhi: Cellobiohydrolase; ment Control Protein; Complement Decay-Accelerating Fac Cellobiohydrolase CeléA; Cellobiohydrolase I: Cellobiohy tor; Complement Factor B: Complement Protein C8Gamma; drolase I Catalytic Domain: Cellobiohydrolase II: Cellobiose Complement Receptor Type 2: COMT (catecol-O-methyl Dehydrogenase; Cellular retinaldehyde-binding protein: Cel transferase); Conantokin-T: Conantoxin G. Congerin I; Con lular retinoic acid binding protein 1: Cellular retinoic acid gerin II: Conjugal Transfer Protein Trwb; Connective Tissue binding protein 2: Cellulase; Cellulase B: Cellulase Cel48F: Activating Peptide-III: Conotoxin Gs; Conserved Hypotheti Cellulase Cel9M; Cellulase Celc: Cellulomonas Fimi Family cal Protein: Conserved Hypothetical Protein Af2008: Con 10 Beta-1,4-Glycana; Cellulosomal Scaffoldin; Cephaibol B; served Hypothetical Protein Mth 1747; Conserved Hypotheti Cephalosporin C Deacetylase; Cephalosporinase; Ceramide cal Protein Tim1158: Conserved Hypothetical Protein glucosyltransferase: Ceruloplasmin; Cg 14704 Protein; Tm 1464; Conserved Hypothetical Protein Ydce; Conserved cGMP Phosphodiesterase: Cgmp Phosphodiesterase A2; Hypothetical Protein Yffb; Conserved Hypothetical Protein Cgmp-Inhibited 3',5'-Cyclic Phosphodiesteras; Cgmp-Spe Yuaa; Conserved Protein Mth 1675; Constitutive Androstane cific 3',5'-Cyclic Phosphodiesterase; Chalcone Synthase: Receptor; Contryphan-R: Contryphan-Sm; Contryphan-Vn, Chalcone Synthase 2: Chalcone-Flavonone Isomerase 1: Major Form: Copper Amine Oxidase; Copper Amine Oxi US 2015/0309021 A1 Oct. 29, 2015 dase; Copper Amine Oxidase, Liver Isozyme; Copper Trans Cytochrome P450 Cyp119; Cytochrome P450 CYP11B1 port Protein Atox1; Copper-Containing Nitrite Reductase: (steroid hydroxylase); Cytochrome P450Cam; Cytochrome Core Protein; Corticosteroid 11-Beta-Dehydrogenase P450-Cam; Cytochrome P450Epok; Cytochrome P450Eryf: Isozyme; Corticosteroid 11-Beta-Dehydrogenase, Isozym; Cytochrome P450Nor; Cytochrome Rc557: Cytoglobin: Corticosteroid 11-beta-dehydrogenase, isozyme 1: Corticos Cytohesin 2: Cytokine Receptor Common Beta Chain; Cyto teroid 11-beta-dehydrogenase, isozyme 2: Corticosteroid kinin Dehydrogenase 1: Cytosine Deaminase; cytosolic Receptor; Corticotropin Releasing Hormone: COX-1: COX 5-nucleotidase II; Cytosolic phospholipase A2; Cytotoxic T 2: Crabp-I: Crca Protein: C-Reactive Protein; Creatine kinase Lymphocyte Associated Antigen 4; Cytotoxic Tcell Valpha B-type: Creatine kinase M-type; Creatine Kinase, B Chain; Domain; Cytotoxin 3: D (1B) dopamine receptor; D (2) Creatine kinase, sarcomeric; Creatine kinase, ubiquitous; Dopamine Receptor; D (3) Dopamine Receptor; D (4) Crk-Associated Substrate; Crotonobetainyl-Coa: Carnitine dopamine receptor, D1 dopamine receptor-interacting pro Coa-Trans; Crotonobetainyl-Coa: Carnitine Coa-Transfera; tein calcyon; D199S Mutant Of Bovine 70 Kilodalton Heat Crustacyanin; Crustacyanin Al Subunit; Crustacyanin C2 Sh; D206S Mutant Of Bovine 70 Kilodalton Heat Sh; D-2- Subunit; Cruzipain; Cryptochrome 1 Apoprotein: Csdb: Csdb Hydroxyisocaproate Dehydrogenase; D3, D2-Enoyl Coa Protein; C-Terminal Analogue Of Neuropeptide Y. A; C-Ter Isomerase Ecil; D-3-Phosphoglycerate Dehydrogenase; minal Binding Protein 1: C-Terminal Binding Protein 3: D-3-Phosphoglycerate Dehydrogenase; Dabd; Dahp Syn C-Terminal Src Kinase; Ctla-4: Ctp Synthase; Ctp: Phospho thetase: D-Ala Ligase: D-Alanine Aminotransferase, D-Ala choline Cytidylyltransferase: C-Type Lectin Dc-Signr, Culti nine: D-Lactate Ligase: D-alanine-D-alanine ligase A: nase; Cyan Fluorescent Protein Cfp; Cyanate Lyase: Cyano D-Alanyl-D-Alanine Carboxypeptidase; D-Amino Acid globin: Cyanovirin-N; Cyclic Alpha Melanocyte Stimulating Aminotransferase: D-Amino Acid Oxidase, D-amino-acid Hormo: Cyclic Hexapeptide Rr; Cyclic Parathyroid Hor oxidase; D-Aminoacylase; Dape; Daptomycin; D-beta-hy mone; Cyclic Phosphodiesterase: Cyclin Dependent Kinase droxybutyrate dehydrogenase, mitochondrial precursor; Subunit, Type 1: Cyclin-Dependent Protein Kinase 2: Cyclo; Dcoh: Ddab: De Novo Designed 21 Residue Peptide; De Cyclodextrin. Glycosyltransferase: Cyclodextrin-Glycosyl Novo Designed Cyclic Peptide; Deacetoxycephalosporin C transferase; Cyclomaltodextrin Glucanotransferase; Synthase: Death-Associated Protein Kinase; Death-Associ Cyclooxygenase-2; Cyclophilin; Cyclopropane-Fatty-Acyl ated Protein Kinase 1: Decorin: Dehaloperoxidase; Dehydro Phospholipid Syn: Cyclopropane-Fatty-Acyl-Phospholipid genase/reductase SDR family member 4: Delta 2 Crystallin; Synthas; Cyclopropane-fatty-acyl-phospholipid synthase 1: Delta Crystallin I; Delta Crystallin II; Delta-1-pyrroline-5- Cyclosporin A. As Bound To Cyclophilin: Cyp175A1: carboxylate dehydrogenase, mitochondrial precursor; Delta CYP1A2: Cystalysin; Cystathionine Beta-Synthase: Cys Aminolevulinic Acid Dehydratase; Delta-Conotoxin Evia; tathionine gamma-lyase, CyStathionine Gamma-Synthase; Deoxycytidine Kinase; Deoxycytidine Triphosphate Deami Cysteine desulfurase: Cysteine dioxygenase fragment; nase; Deoxycytidylate Deaminase; Deoxycytidylate Cysteine dioxygenase type I: Cysteine Sulfinic acid decar Hydroxymethylase; Deoxy-D-Mannose-Octulosonate boxylase: Cysteine-Rich Domain Of Mannose Receptor; 8-Phosphate Pho; Deoxyhypusine Synthase; Deoxyhypusine Cysteinyl leukotriene Receptor 1: Cysteinyl leukotriene Synthase: Deoxynucleoside Monophosphate Kinase; Deox receptor 2: Cysteinyl-tRNA Synthetase: Cystic Fibrosis yribodipyrimidine Photolyase; Deoxyribonuclease I: Deox Transmembrane Conductanc: Cystic Fibrosis Transmem yribonucleoside Kinase; Deoxyribose-Phosphate Aldolase; brane Conductance Re; Cystine/glutamate transporter; Cys Deoxyuridine 5'-Triphosphate Nucleoditohydro; Deoxyuri tinosin; Cytidine Deaminase; Cytidine Monophospho-N- dine 5'-Triphosphate Nucleotide Hydr; Deoxyuridine Acetylneuraminic Acid; Cytidylate Kinase; Cytochrome 5'-Triphosphate Nucleotidohydro: Deoxyuridine Triphos B=5=Reductase; Cytochrome B2; Cytochrome B2, Mito phatase; Dephospho-Coa Kinase; Der F II: DesGlyl chondrial; Cytochrome B5; Cytochrome B5 Outer Mitochon Contryphan-R: Designed Protein Ctpr2: Designed Protein drial Membrane Is; Cytochrome B562; Cytochrome C: Cyto Ctpr3: Dethiobiotin Synthetase; Devb Protein: Dextranase; chrome C'; Cytochrome C"; Cytochrome C Family Protein; D-Galactose)-GLUCOSE BINDING PROTEIN; D-Glu Cytochrome C Nitrite Reductase; Cytochrome c oxidase sub cose 6-Phosphotransferase: D-Glyceraldehyde-3-Phosphate unit 1; Cytochrome CPeroxidase; Cytochrome CPeroxidase, Dehydrogena: D-Glyceraldehyde-3-Phosphate Dehydroge Mitochondrial; Cytochrome C. Iso-1; Cytochrome C. Puta nase, D-Glyceraldehyde-3-Phosphate-Dehydrogena; tive: Cytochrome C2; Cytochrome C2, Iso-2; Cytochrome D-Glyceraldehyde-3-Phosphate-Dehydrogenase; Dhps, C3: Cytochrome C3, A Dimeric Class III C-Type Cy; Cyto Dihydropteroate Synthase: D-Hydantoinase; Diacylglycerol chrome C4; Cytochrome C549; Cytochrome C550; Cyto kinase; Diadenosine Tetraphosphate Hydrolase; Diamine chrome C551; Cytochrome C-551; Cytochrome C551 Per acetyltransferase 1; Diamine acetyltransferase 2: Diami oxidase; Cytochrome C552; Cytochrome C-552; nopimelate Decarboxylase; Dianthin 30; Dienelactone Cytochrome C-553; Cytochrome C-554; Cytochrome C-556; Hydrolase; Dienoyl-Coa Isomerase; Diga16: Di-Haem Cyto Cytochrome C6: Cytochrome C7: Cytochrome Cd1 Nitrite chrome C Peroxidase; Diheme Cytochrome C Napb: Di Reductase; Cytochrome Cl; Cytochrome F: Cytochrome Heme Peroxidase; Dihydrodipicolinate Reductase; Dihy Oxidase Subunit II: Cytochrome P450; Cytochrome P450 drodipicolinate Reductase; Dihydrodipicolinate Synthase: 107A1; Cytochrome P450 1 19; Cytochrome P450 121; Cyto Dihydrofolate Reductase; Dihydrofolate Reductase (ma chrome P450 152A1; Cytochrome P450 154C1; Cytochrome larial); Dihydrolipoamide Dehydrogenase; Dihydrolipoyl P450 17A1; Cytochrome P450 19 (Aromatase); Cytochrome dehydrogenase, mitochondrial precursor, Dihydrolipoyl P450 27: Cytochrome P450 2B4; Cytochrome P450 2C5; Transacetylase; Dihydrolipoyllysine-residue acetyltrans Cytochrome P4502C8: Cytochrome P450 2C9; Cytochrome ferase component of pyruvate dehydrogenase complex, mito P450 2R1; Cytochrome P450 3A4; Cytochrome P450 4A11 chondrial precursor; Dihydroneopterin Aldolase; Dihydro precursor; Cytochrome P450 51; Cytochrome P450 51 orotase; Dihydroorotate Dehydrogenase; Dihydroorotate (Yeast); Cytochrome P450 55A1; Cytochrome P450 Bm-3: Dehydrogenase A; Dihydroorotate Dehydrogenase, mito US 2015/0309021 A1 Oct. 29, 2015 46 chondrial Precursor: dihydropterate synthase (bacterial); Beta-Xylanase; Endo-1,4-beta-Xylanase 2 precursor, Endo Dihydropteridine Reductase: Dihydropteridine Reductase: 1,4-Beta-Xylanase A: Endo-1,4-Beta-Xylanase A Precursor: Dihydropteroate synthase: Dihydropteroate Synthase (ma Endo-1,4-Beta-Xylanase II: Endo-1,4-Beta-Xylanase Y: larial); Dihydropteroate synthase (Pneumocystis carinii); Endo-Alpha-Sialidase; Endo-Beta-1,4-Glucanase; Endo Dihydropteroate Synthase I: Dihydropyridine calcium chan Beta-N-Acetylglucosaminidase F3; Endocellulase E 1: Endo nel; Dihydropyridine-sensitive L-type, calcium channel glucanase; Endoglucanase 5A: Endoglucanase 9G. Endoglu alpha-2/ delta Subunits; Dihydropyrimidine dehydrogenase; canase A: Endoglucanase B; Endoglucanase C. Dihydroxyacetone Kinase; Diisopropylfluorophosphatase; Endoglucanase I: Endoglucanase I: Endoglucanase V Cello Dimeric Hemoglobin: Dimethyl Sulfoxide Reductase: biose Complex: Endoplasmic Reticulum Mannosyl-Oli Dipeptidyl Aminopeptidase-Like Protein 6: Dipeptidyl Pep gosacchari; Endoplasmin, Endopolygalacturonase; Endopo tidase I: Dipeptidyl Peptidase IV: Dipeptidyl Peptidase IV: lygalacturonase; EndoSome-Associated Protein, Endothelial Dipeptidyl Peptidase IV Soluble Form; Diphtheria Toxin; Nitric-Oxide Synthase: Endothelial Protein C Receptor; Diphtheria Toxin Repressor; Diphthine Synthase: Dissimila Endothelin B receptor precursor; Endothelin-1; Endothelin-1 tory Copper-Containing Nitrite: Dissimilatory Copper-Con receptor precursor, Endothiapepsin, Endothiapepsin precur taining Nitrite Redu; D-lactate dehydrogenase: D-Lactate sor; Endoxylanase; Endoxylanase 1 1A: Engrailed Home Dehydrogenase; Dip-1: D-Maltodextrin-Binding Protein; odomain; Enhancing Lycopene Biosynthesis Protein; Dmso Reductase; DNA Adenine Methylase; DNA Beta-Glu Enkephalinase; Enolase; Enolase 1: Enoyl Acp Reductase: cosyltranferase: DNA Cytosine Methyltransferase Dnmt2: Enoyl Acyl Carrier Protein Reductase; Enoyl-Acyl-Carrier DNA Double-Strand Break Repair RadS0 Atpase; DNA Protein Reductase: Enoyl-Acyl-Carrier-Protein Reduc Gyrase B; DNA gyrase subunit A; DNA Gyrase Subunit B; tase; Enoyl-Acyl-Carrier-Protein Reductase: Enoyl-Acyl DNA Helicase; DNA Ligase; DNA Ligase I: DNA Ligase III; Carrier-Protein Reductase Nadh: Enoyl-acyl-carrier-pro DNA Ligase, Nad-Dependent; DNA Ligase, Nad-Depen tein reductase NADH: Enoyl-Acp Reductase: Enoyl-Acp dent; DNA Mismatch Repair Protein Mutl; DNA Nucleotide Reductase: Enoyl-Acyl Carrier Protein: Enoyl-Coa Excision Repair Enzyme Uvrb; DNA Photolyase; DNA Poly Hydratase: Enoyl-Coa Hydratase, Mitochondrial; Envelope merase; DNA Polymerase (human cytomegalovirus); DNA Glycoprotein; Envelope Glycoprotein; Envelope glycopro polymerase (human herpes simplex virus 1); DNA Poly tein GP340; Envelope glycoprotein GP340/GP220; Eosino merase (human); DNA polymerase (Varicella-zoster virus); phil Cationic Protein; Eosinophil Lysophospholipase Chain: DNA Polymerase alpha (human); DNA Polymerase Beta; A: Eosinophil-Derived Neurotoxin; Ep-Cadherin; Ephrin DNA Polymerase I: DNA Polymerase III Subunit Gamma; Type-A Receptor 2; Ephrin-A5; Ephrin-B2: Epidermal DNA Polymerase III, Epsilon Chain; DNA Primase; DNA Growth Factor: Epidermal Growth Factor Receptor; Epider Primase Small Subunit; DNA Primase/Helicase; DNA Repair mal Growth Factor Receptor 2: Epidermal growth factor and Recombination Protein Rad; DNA Repair and Recombi receptor precursor: Epidermin Modifying Enzyme Epid; Epi nation Protein Rada; DNA Repair Protein Xrcc4; DNA Rep didymal Secretory Protein E1; Epilancin 15X; Epoxide lication Protein; DNA Topoisomerase I: DNA Topoisomerase Hydrolase: Epoxide Hydrolase 2, Cytoplasmic; Epsin; Epsp II; DNA Topoisomerase II (bacterial); DNA-3-Methylad Synthase: Ergosterol biosynthetic protein 28; Ermc' Methyl enine Glycosylase I: DNA-3-Methyladenine Glycosylase II; transferase: Erv2 Protein, Mitochondrial; Erythrina Crista DNA-Binding Response Regulator; DNA-directed RNA Galli Lectin; Erythropoietin receptor, Esal Histone Acetyl polymerase (E. coli); DNA-directed RNA polymerase beta transferase; Esal Protein: E-Selectin; Esta; Esterase; chain; DNA-directed RNA polymerase beta' chain; DNA Estradiol 17 Beta-Dehydrogenase 1: Estradiol 17 Beta-De Directed RNA Polymerase Subunit L: Dodecin. DOPA hydrogenase 4: Estradiol 17-beta-dehydrogenase 1: Estradiol Decarboxylase; Dopamine beta hydroxylase; Dopamine D1 17-beta-dehydrogenase 2: Estradiol 17-beta-dehydrogenase Receptor, Dopamine reuptake pump; Dp-Tt2; Dr Hemagglu 3: Estradiol 17-beta-dehydrogenase 8: Estrogen Receptor; tinin Structural Subunit; D-Ribose-5-Phosphate Isomerase: Estrogen Receptor (ER); Estrogen Receptor Alpha; Estrogen D-Ribose-Binding Protein Complexed With Beta; D-Ribose Receptor Beta; Estrogen Sulfotransferase; Estrogen-Related Binding Protein Mutant With Gly 134; D-Ribose-Binding Receptor Gamma; Eukaryotic Peptide Chain Release Factor Protein Mutant With IIe 132: Drosophila Neuroglian; Dtdp GTP-, Eukaryotic Translation Initiation Factor 4E: Evolved 4-Dehydrorhamnose 3.5-Epimerase: Dtdp-4-Dehydrorham beta-galactosidase alpha-Subunit; Excinuclease Abc Subunit nose Reductase, Rfbd 0; Dtdp-6-Deoxy-D-Xylo-4-Hexulose B; Exo-; Exocellobiohydrolase I: Exoglucanase I: Exo-lnuli 3.5-Epimerase: Dtdp-D-Glucose 4,6-Dehydratase; Dtdp nase; Exo-Maltotetraohydrolase; Exopolyphosphatase; Exo Glucose Oxidoreductase: Dual Adaptor Of Phosphotyrosine toxin A: Expressed Protein; Exterior Membrane Glycopro and 3-Phosp: Dual Specificity Mitogen-Activated Protein K: tein Gp120; Extracellular calcium-sensing receptor Dual Specificity Protein Kinase Clk1; Duck Ovotransferrin; precursor: Extracellular Regulated Kinase 2: Extracellular Duodenase; Dutp Pyrophosphatase: D-Xylose Isomerase: E. Signal-Regulated Kinase 2: Extracellular Subtilisin-Like coli citrate synthetase; E. coli glutathione reductase; E. coli Serine Protein: Extrahepatic lipoprotein lipase (LL); F17-Ag malate dehydrogenase; E. Coli Maltodextrin Phosphorylase; Lectin; F1-Gramicidin A; F1-Gramicidin C: F420-Depen E. coli pyruvate dehydrogenase, E. coli ribosomal proteins; dent Alcohol Dehydrogenase; F65A/Y131C-Mi Carbonic Eafp 2: Early Endosomal Autoantigen 1; Ecotin: Eh Domain Anhydrase V: Factor D: Factor IIIFactor IX Factor VIIIFactor Binding Protein Epsin; Eif2Gamma; Elastase; Elastase 1: X; Factor Inhibiting Hifl; Farnesyl Diphosphate Synthase: Eledoisin; Elongation Factor: Elongation Factor 1-Alpha; Farnesyl pyrophosphate synthetase; Fasciclin I; Fatty Acid Elongation Factor 2: Elongation Factor G; Elongation Factor Binding Protein; Fatty Acid Binding Protein Homolog: Fatty G Complexed With Guanosine; Elongation Factor Tu; Endo/ Acid Metabolism Regulator Protein; Fatty Acid/Phospho Exocellulase E4, Endo-1,4-B-D-Mannanase; Endo-1,4-Beta lipid Synthesis Protei: Fatty Acid-Binding Protein; Fatty Glucanase Engf; Endo-1,4-Beta-D-Xylanase; Endo-1,4- Acid-Binding Protein, Adipocyte; Fatty Acid-Binding Pro Beta-Glucanase; Endo-1,4-Beta-Glucanase F. Endo-1,4- tein, Brain; Fatty acid-binding protein, liver; Fatty aldehyde US 2015/0309021 A1 Oct. 29, 2015 47 dehydrogenase; Fatty-Acid Amide Hydrolase. F-Box Only aminobutyric-acid receptor alpha-3 Subunit precursor; Protein 2: Fc Fragment; Fc Gamma Receptor FCGR1 HU Gamma-aminobutyric-acid receptor alpha-4 Subunit precur MAN: Feglymycin; Feline Immunodeficiency Virus Pro sor; Gamma-aminobutyric-acid receptor alpha-5 Subunit pre tease: Feline Leukemia Virus Receptor-Binding Domai; cursor; Gamma-aminobutyric-acid receptor alpha-6 Subunit Ferredoxin; Ferredoxin II; Ferredoxin Reductase; Ferre precursor; Gamma-aminobutyric-acid receptorrho-1 Subunit doxin: Nadp+Oxidoreductase: Ferredoxin: Nadp+Reduc Precursor: Gamma-Glutamyl Hydrolase; Ganglioside Gm2 tase; Ferredoxin-Dependent Glutamate Synthase: Ferre Activator; Gastrotropin; GDH/6PGL endoplasmic bifunc doxin-Nadp Reductase: Ferredoxin-Nadip Reductase: tional protein precursor Includes: Glucose 1-dehydroge Ferredoxin-Nadp+Reductase; Ferredoxin-Nadp+Reductase: nase; Gdnf Family Receptor Alpha 1: GDP-D-Mannose-4,6- Ferric Hydroxamate Receptor; Ferric Hydroxamate Uptake Dehydratase; GDP-Fucose Synthetase; GDP-L-fucose Receptor, Ferrichrome-Binding Periplasmic Protein; Fer synthetase; GDP-Mannose 4.6 Dehydratase; GDP-Mannose richrome-lron Receptor; Ferrichrome-lron Receptor Precur 4,6-Dehydratase; GDP-Mannose 6-Dehydrogenase; GDP sor; Ferripyochelin Binding Protein: Ferripyoverdine Recep Mannose Mannosyl Hydrolase; General Control Protein tor; Ferritin heavy chain; Ferritin light chain; Ferrochelatase: Gcn4; General Secretion Pathway Protein E: General Stress Feruloyl Esterase A; Fez-1 Beta-Lactamase: Fgf Receptor 1: Protein 69; Genome Polyprotein; Gephyrin; Geranyltran Fiber Protein: Fibrin; Fibrinogen-420; Fibroblast Activation stransferase; Gfp-Like Chromoprotein Fp595; Gial; Giding Protein, Alpha Subunit; Fibroblast Growth Factor 9: Fibro Protein-Mglb: Glandular Kallikrein-13; Glcnac1P Uridyl blast Growth Factor Receptor 2: Fibroblast growth factor-4 transferase Isoform 1: Agx1; Globin; Globin I; Globin Li637; precursor: Fibronectin: Fimbrial Lectin; Fk506 Binding Pro Globin-3; Glpe Protein; Glucagon receptor; Glucan 1,3-Beta tein; Fk506-Binding Protein; FK506-binding protein 1A: Glucosidase I/II: Glucan 1,4-Alpha-Maltohexaosidase; Glu Fk506-Binding Protein 4; Fkbp12.6; FKBP12-rapamycin carate Dehydratase; Glucoamylase; Glucoamylase-471; Glu complex-associated protein; Fkbp25; Fkbp-Type Peptidyl cocorticoid Receptor, Glucokinase; Glucokinase Isoform 2: Prolyl Cis-Trans Isom; Fksg76; FL cytokine receptor precur Gluconate 5-Dehydrogenase; Gluconate Kinase; Glu sor: Flavin reductase: Flavocytochrome B2: Flavocyto cosamine 6-Phosphate Synthase; Glucosamine-6-Phosphate chrome C: Flavocytochrome C Fumarate Reductase: Deaminase; Glucosamine-6-Phosphate Isomerase: Glu Flavocytochrome C3; Flavocytochrome C3 Fumarate Reduc cosamine-Fructose-6-Phosphate Aminotrans; Glucosamine tase: Flavodoxin; Flavodoxin Reductase: Flavohemoprotein; Phosphate N-Acetyltransferase: Glucose 1-Dehydrogenase; Flavoprotein; Fluorescent Protein Fp538; Fmn-Binding Pro Glucose 6-Phosphate 1-Dehydrogenase; Glucose 6-Phos tein: Fms1 Protein: Focal Adhesion Kinase 1: Folate receptor phate 1-Dehydrogenase; Glucose 6-Phosphate Dehydroge alpha; Folate receptor beta; Folate receptor gamma; Folate nase; Glucose Dehydrogenase; Glucose Oxidase; Glucose-1- transporter 1: Folc Bifunctional Protein; Follicle Stimulating Phosphatase: Glucose-1-Phosphate Adenylyltransferase Hormone Receptor; Follistatin: Folylpolyglutamate syn Smal; Glucose-1-Phosphate Cytidylyltransferase: Glucose thase; Folylpolyglutamate synthase, mitochondrial; 1-Phosphate Thymidylyltransferas: Glucose-1-Phosphate Folylpolyglutamate Synthetase: Formaldehyde Dehydroge Thymidylyltransferase: Glucose-6-Phosphate 1-Dehydroge nase; Formaldehyde Ferredoxin Oxidoreductase: Formalde nase; Glucose-6-Phosphate Isomerase: Glucose-Fructose hyde-Activating Enzyme Fae; Formate Acetyltransferase 1: Oxidoreductase; Glucose-Fructose Oxidoreductase; Glu Formate Dehydrogenase H; Formiminotransferase-Cy cose-Resistance Amylase Regulator, Glucosylceramidase; clodeaminase; Formyl-Coenzyme A Transferase: Formylme Glucuronyltransferase I: Glutaconyl-Coa Decarboxylase A thionine Deformylase: Four-Helix Bundle Model; Fpra; Fr-1 Subunit; Glutamate NMDA receptor subunit epsilon 1 pre Protein; Fragile Histidine Protein; Fragile Histidine Triad cursor; Glutamate NMDA receptor subunit epsilon 2 pre Protein; Fructan 1-Exohydrolase Iia; Fructose 1,6-Bisphos cursor; Glutamate NMDA receptor subunit epsilon 3 pre phatase. Fructose 1,6-Bisphosphatase/Inositol Monopho: cursor; Glutamate Carboxypeptidase II: Glutamate Fructose 1,6-Bisphosphate Aldolase; Fructose-1,6-Bis: Fruc decarboxylase 1: Glutamate decarboxylase 2: Glutamate tose-1,6-Bisphosphatase; Fructose-2,6-Bisphosphatase: Decarboxylase Alpha; Glutamate Decarboxylase Beta; Fructose-Bisphosphate Aldolase; Fructose-Bisphosphate Glutamate Dehydrogenase; Glutamate Dehydrogenase; Aldolase A. Fructose-Bisphosphate Aldolase Class I; Fruc Glutamate Dehydrogenase 1: Glutamate dehydrogenase 1, tose-Bisphosphate Aldolase II; FtsZ. Fucose-Specific Lectin; mitochondrial precursor; Glutamate dehydrogenase 2, mito Fumarase C; Fumarate Hydratase Class II; Fumarate reduc chondrial precursor, Glutamate Racemase; Glutamate recep tase flavoprotein subunit; Fumarylacetoacetate Hydrolase; tor 1 Precursor: Glutamate Receptor 2: Glutamate Receptor Fusion Protein; Fusion Protein Consisting Of Kinesin-Like 2 Precursor; Glutamate receptor 3: Glutamate receptor 4: Pr; Fusion Protein Consisting Of Staphylococcus: Fv Frag Glutamate Receptor 6: Glutamate Receptor Subunit 2: ment; G Protein Gi Alpha 1: G15-Gramicidin A; G25K GTP Glutamate Receptor, Ionotropic Kainate 1: Glutamate recep Binding Protein; GABA Transaminase; GABAA Receptor; tor, ionotropic kainate 1 precursor; Glutamate Receptor, GABA-B Receptor; Gag Polyprotein; Ga110 Bifunctional Ionotropic Kainate 2: Glutamate receptor, ionotropic kainate Protein; Galactanase; Galactokinase; Galactose Mutarotase; 3: Glutamate receptor, ionotropic kainate 5: Glutamate Semi Galactose Oxidase; Galactose Oxidase Precursor; Galactose aldehyde Aminotransferase; Glutamate-Cysteine Ligase; 1-Phosphate Uridyl Transferase-Lik; Galactose-1-Phosphate Glutamate-cysteine ligase catalytic subunit: Glutamate-cys Uridylyltransferase; Galactose-Binding Protein Complex teine ligase regulatory subunit; Glutaminase, kidney isoform; With Gluco; Galactose-Specific Lectin; Galactoside Glutaminase, liver isoform; Glutaminase-Asparaginase; O-Acetyltransferase; Galactosylgalactosylxylosylprotein Glutamine Aminotransferase: Glutamine Phosphoribosylpy 3-Beta-G: Galectin-1; Galectin-1; Galectin-2: Galectin-3, rophosphate: Glutamine Phosphoribosylpyrophosphate Ami Galectin-3; Galectin-7: Gamma Chymotrypsin; Gamma dot; Glutamine Receptor 2: Glutamine Synthetase: Aminobutyrate Metabolism Dehydratase/I: Gamma-ami Glutamyl-Endopeptidase; Glutamyl-tRNA Reductase: nobutyric-acid receptor alpha-2 Subunit precursor; Gamma Glutamyl-tRNA Synthetase: Glutaredoxin 3: Glutaryl-Coa US 2015/0309021 A1 Oct. 29, 2015 48

Dehydrogenase; Glutathine Synthetase; Glutathione Reduc ing Nuclear Protein Ran; GTP-Binding Protein Ran; GTP tase; Glutathione reductase (mitochondrial); Glutathione Binding Protein Rheb; GTP-Binding Protein YptS1; GTP S-Transferase; Glutathione S-Transferase; Glutathione Binding Protein Ypt7P; GTP-Binding Protein Ysxc: S-Transferase 1-6: Glutathione S-Transferase 2: Glutathione Guanidinoacetate Methyltransferase; Guanidinoacetate S-Transferase 26 Kda; Glutathione S-Transferase A1: Glu N-Methyltransferase; Guanine Deaminase; Guanine Nucle tathione S-Transferase A1-1; Glutathione S-Transferase otide Exchange Factor and I; Guanine Nucleotide Exchange A3-3; Glutathione S-Transferase Class Pi Chimaera; Glu Factor and Integ; Guanine Nucleotide-Binding Protein G: tathione S-Transferase Gita-1A; Glutathione S-Transferase Guanine Phosphoribosyltransferase; Guanylate Kinase; Gua Mu 1. Glutathione S-Transferase Mu 2: Glutathione S-Trans nyl-Specific Ribonuclease T1: Guanyl-Specific Ribonu ferase P. Glutathione S-transferase pi; Glutathione S-Trans clease T1 Precursor; Gurmarin; H-; H. Pylorirdx A: H+/K+ ferase Tsi-1, Glutathione S-Transferase Ya Chain; Glu ATPase (Proton pump); Hal: Haematopoetic Cell Kinase; tathione S-Transferase Yb 1: Glutathione S-Transferase Yfyf: Haloalkane Dehalogenase; Halohydrin Dehalogenase; Glutathione S-Transferase, Mitochondrial; Glutathione Syn Halorhodopsin; Halotolerance Protein Hal2: Halotolerance thetase; Glutathione Transferase; Glutathione Transferase; Protein Hal 3: Hev NS5B Polymerase: Heys Beta3-Cys Ana Glutathione Transferase Gst 1-3. Glutathione Transferase logue Of Hiv Gp41; Halp; Head Decoration Protein: Heat Gst1-6: Glutathione Transferase Zeta; Glutathione-Depen Shock 70 Kda Protein 1; Heat Shock 90Kda Protein 1, Alpha; dent Formaldehyde Dehydroge: Glutathione-Dependent Heat Shock Cognate Kda70; Heat Shock Locus U; Heat Formaldehyde-Activatin: Glutathione-Requiring Prostaglan Shock Protein 90; Heat Shock Protein Hslu; Heat Shock din D Syntha; Glutathione-S-Transferase; Glyceraldehyde Protein Hsp90-Alpha; Heat Shock Protein Hsp90-Beta; Heat 3-Phosphate Dehydrogenase; Glyceraldehyde 3-Phosphate Shock Protein Hsp33; Heat Shock Transcription Factor; Heat Dehydrogenase; Glyceraldehyde 3-Phosphate Dehydroge Shock-Like Protein 1: Heat-labile enterotoxin B chain pre nase A; Glyceraldehyde-3-Phosphate Dehydrogenase; Glyc cursor; Heat-Labile Enterotoxin B Subunit; Heat-Shock 70 eraldehyde-3-Phosphate Dehydrogenase; Glyceraldehyde-3- Kd Protein; Heat-Shock Cognate 70 Kd Protein: Heavy Phosphate Dehydrogenase A: Glyceraldehyde-3-phosphate chain; Heavy chain 1 B72.3 (murine); Hemagglutinin Precur dehydrogenase, liver; Glyceraldehyde-3-phosphate dehydro Sor, Hemagglutinin-Neuraminidase; Hemagglutinin genase, testis-specific; Glycerol Dehydratase; Glycerol Neuraminidase Glycoprotein; Hematopoetic Cell Kinase Dehydrogenase; Glycerol Kinase; Glycerol Uptake Facilita Hck; Hematopoietic Prostagladin D Synthase: Heme Oxyge tor Protein; Glycerol Uptake Operon Antiterminator-Re: nase; Heme Oxygenase 1; Heme Oxygenase 2: Heme Oxy Glycerol-3-Phosphate Cytidylyltransferase: Glycerol-3- genase-1; Heme Pas Sensor Protein: Heme-Based Aerotactic Phosphate Dehydrogenase; Glycerol-3-phosphate dehydro Transducer Hemat; Heme-Based Methyl-Accepting Chemo genase NAD+, cytoplasmic; Glycinamide ribonucleotide taxis P: Heme-Based Methyl-Accepting Chemotaxis Prote: formyltransferase (fragment); Glycinamide Ribonucleotide Heme-Binding Protein A: Hemerythrin; Hemk Protein; Transformylase; GLycine alpha 2 receptor, Glycine amidi Hemo; Hemocyanin; Hemocyanin; Hemoglobin: Hemoglo notransferase; Glycine Betaine-Binding Periplasmic Protein; bin beta chain; Hemoglobin Gamma Chains; Hemoglobin V: Glycine dehydrogenase; Glycine N-Methyltransferase: Gly Hemoglobin-Like Protein Hbn; Hemoglobin-Like Protein cine Oxidase; Glycine receptor alpha-1 chain Precursor: Hbo: Hemolytic Lectin Cel-III: Hemolytic Lectin Lsla: Glycine receptor alpha-3 chain; Glycine receptor beta chain; Hemopexin; Heparan Sulfate; Heparan Sulfate D-Glu Glycogen phosphorylase; Glycogen Phosphorylase b: Gly cosaminyl 3-O-Sulfotra: Heparan Sulfate N-Deacetylase/N- cogen Phosphorylase, Liver Form; Glycogen phosphorylase, Sulfotransfe; Heparin Binding Protein; Heparin Cofactor II; muscle form; Glycogen Synthase 1: Glycogen Synthase Heparin cofactor II precursor; Heparin-Binding Growth Fac Kinase-3 Beta; Glycogenin-1; Glycolate Oxidase; Glycolipid tor 1; Heparin-binding growth factor 1 precursor; Heparin 2-Alpha-Mannosyltransferase: Glycolipid Transfer Protein; Binding Protein: Hepatitis C Virus Ns5B RNA Polymerase: Glycoprotein D: Glycoprotein-Fucosylgalactoside Alpha Hepatitis C Virus Ns5B RNA-Dependent RNA Pol; Hepato Galac; Glycoprotein-Fucosylgalactoside Alpha-N-Ace; Gly cyte Growth Factor; Hepatocyte Growth Factor Activator Pre cosyl Transferase: Glycosylase; Glycosyltransferase A: Gly cursor; Hepatocyte Growth Factor Receptor; Hepatocyte cosyltransferase B; Glycosyltransferase Gtfa; Growth Factor-Regulated Tyrosine; Hepatocyte Nuclear Fac Glycosyltransferase Gitfl; Glyoxalase Family Protein; Gly tor 1-Alpha; Hepatocyte Nuclear Factor 4-Alpha; Hepatocyte oxalase II: Glyoxylate reductase/hydroxypyruvate reductase; Nuclear Factor 4-Gamma; Her-1 Protein; Heroin Esterase; Gmp Reductase I: GMP synthase glutamine-hydrolyzing: Herpes Thymidine Kinase; Hevamine: Hevamine A: Hevea Gmp Synthetase; Gomesin; Gonadotropin-releasing hor brasiliensis; Hevein; Hevein: Hexokinase: Hexokinase Type mone II receptor, Gonadotropin-releasing hormone receptor, I: Hexon Protein: Hexose-1-Phosphate Uridylyltransferase: GP41 envelope protein (first heptad repeat); Gp70; GPIIb Hi0065; Hil317; High Affinity Immunoglobulin Epsilon Receptor; GPIIIa Receptor; Gramicidin; Gramicidin A; Recepto: High affinity immunoglobulin epsilon receptor Gramicidin B; Gramicidin C. Gramicidin D: Gramicidin alpha-Subunit precursor, High affinity immunoglobulin epsi Synthetase 1: Granulocyte colony stimulating factor receptor lon receptor gamma-Subunit precursor, High Affinity Ribose (CD114 antigen); Granulocyte-macrophage colony Stimulat Transport Protein Rbsd; High Mobility Group Protein 1: ing factor receptor (GM-CSF-R-alpha or CSF2R); Granul High-Affinity Branched Chain Amino Acid Tran; High-Af ysin; Granzyme B; Green Fluorescent Protein; Green-Fluo finity Branched-Chain Amino Acid Tran; High-affinity rescent Protein; Griffonia Simplicifolia Lectin 4: Group X cAMP-specific 3',5'-cyclic phosphodiesterase 7A: High-af Secretory Phospholipase A2: Growth Differentiation Factor finity cationic amino acid transporter-1; High-affinity choline 5: Growth Factor Receptor-Bound Protein 2: Growth hor transporter 1; High-Molecular-Weight Cytochrome C. Hista mone releasing hormone receptor (GRF receptor); Growth mine H1 Receptor; Histamine H2 Receptor; Histamine H4 arrest-specific protein 6: Grp 1: Gst2 Gene Product; GTP Receptor. Histamine N-Methyltransferase: Histidine ammo Cyclohydrolase I: Gtpase-Activating Protein 1: GTP-Bind nia-lyase. Histidine Containing Protein; Histidine decar US 2015/0309021 A1 Oct. 29, 2015 49 boxylase: Histidine Triad Nucleotide-Binding Protein: Histi Rv1170; Hypothetical Protein Rv1347C/Mt1389: Hypotheti dine triad nucleotide-binding protein 1: Histidinol cal Protein Rv2238C/Mt2298: Hypothetical Protein Rv2991; Dehydrogenase. Histidinol Phosphate Aminotransferase: Hypothetical Protein Slr1257: Hypothetical Protein Smu. Histidinol-Phosphate Aminotransferase: Histidyl-tRNA syn 260: Hypothetical Protein Sso2532: Hypothetical Protein thetase: Histo-Blood Group Abo System Transferase: His Ta0175: Hypothetical Protein Tal320: Hypothetical Protein tone Acetyltransferase Gcn5; Histone Deacetylase 8: Histone Tm()021: Hypothetical Protein Tm()449; Hypothetical Pro H3; Histone Methyltransferase Doti L; Histone-Lysine tein Tm1070; Hypothetical Protein Tm1380: Hypothetical N-Methyltransferase, H3 Lysin; Hiv 1 Gp41 Hser Analogue Protein Tm 1457: Hypothetical Protein Tm 1553: Hypotheti Peptide Ace-IIe-T: Hiv-1 Integrase: Hiv-1 Protease: HIV-1 cal Protein Tm 1643; Hypothetical Protein Tm841; Hypo Reverse Transcriptase; HIV-2 Protease; Hmg-Coa Reduc thetical Protein Tt0907: Hypothetical Protein Tt 1426; Hypo tase; Holliday Junction DNA Helicase Ruvb; Holliday-Junc thetical Protein Vc1899: Hypothetical Protein Vca0042: tion Resolvase: Holo-; Holo-D-Glyceraldehyde-3-Phosphate Hypothetical Protein Ycdx: Hypothetical Protein Ycfc: Dehydrogen; Homo Sapiens V-Kit Hardy-Zuckerman 4 Hypothetical Protein Ydce: Hypothetical Protein Yddu: Feline; Homoprotocatechuate 2,3-Dioxygenase; Hypothetical Protein Yese: Hypothetical Protein Yfdw: Homoserine Dehydrogenase; Homoserine Kinase; Hormone Hypothetical Protein Ygbm; Hypothetical Protein Yhai; Receptor Alpha 1. Thra1; Horseradish Peroxidase C1A: Host Hypothetical Protein Yhda: Hypothetical Protein Yhfp: Factor For Q Beta; Hpha; H-Protein; HpV11 Regulatory Pro Hypothetical Protein, Similar To Potassium C. Hypothetical tein E2: Hst2 Protein: HTH-type transcriptional regulator Protein, Similar To Strepto: Hypothetical Shikimate 5-Dehy malT: Htiv-1 Gp21 Ectodomain/Maltose-Binding Prote: drogenase-L; Hypothetical Transcriptional Regulator I; Human Beta2-Glycoprotein I; Human Cd2; Human Cyto Hypothetical Transcriptional Regulator In Qa: Hypothetical chrome P450 CYP2D6; Human Cytochrome P450 CYP3A4; tRNA/Rrna Methyltransferase HiO: Hypothetical Upf)124 Human Growth Hormone Receptor; Human Immunodefi Protein Yfilh; Hypothetical Upfol31 Protein pH0828: Hypo ciency Virus Type 2: Human keratinocyte growth factor thetical Upfo204 Protein Af0625; Hypothetical Zinc-Type receptor; Human Neutrophil Gelatinase: Human Ornithine Alcohol Dehydrogenase; Hypoxanthine Phosphoribosyl Decarboxylase; Human Procathepsin L. Human Protective transferase: Hypoxanthine-Guanine Phosphoribosyltrans Protein; Human RSV Fusion glycoprotein; Human Sigma fera; Hypoxanthine-Guanine Phosphoribosyltransfera; Alcohol Dehydrogenase; Human Sorbitol Dehydrogenase; hypoxanthine-guanine phosphoribosyltransferase; Hypox Human Thioltransferase: Human Thioredoxin Peroxidase-B; anthine-Guanine-Xanthine Phosphoribosyl: lag-Nucleoside Hut Operon Positive Regulatory Protein: Hyaluronate Lyase: Hydrolase; IcIr Transcriptional Regulator: Ig Gamma-1 Hyaluronate lyase precursor, Hyaluronidase; Hyaluronoglu Chain C Region; Ig Gamma-2A Chain C Region, Ig kappa cosaminidase: Hybrid; Hybrid Cluster Protein: Hydantoin chain V-III region GOL: IgA2: Igf-1 Receptor Kinase; IGG1; ase; Hydrogen peroxide-inducible genes activator; Hydro II Purple Acid Phosphatase; 11-6 Receptor Alpha Chain; IIeal lase; Hydrolase Angiogenin, Hydroxyacid Oxidase 3: Lipid Binding Protein: Imaginal Disc Growth Factor-2: Imi Hydroxyacylglutathione Hydrolase; Hydroxyethylthiazole dazole Glycerol Phosphate Dehydratase: Imidazole Glycerol Kinase; Hydroxylamine Oxidoreductase; Hydroxylamine Phosphate Synthase Hishf Immunoglobulin Alpha Fc Reductase: Hydroxymethylglutaryl-CoA lyase; Hydroxyni Receptor. Immunoglobulin Heavy Chain Epsilon-1. Immu trile Lyase, Hydroxyquinol 1,2-Dioxygenase, HydroxyS noglobulin Lambda Light Chain; Immunoglobulin Lambda teroid Sulfotransferase: Hypothetical 22.5 Kda Protein. In Light Chain Dimer, Immunoglobulin Vh Domain; Immuno Tub1-Cp; Hypothetical 22.5 Kda Protein. In Tub1-Cpr3 I: globulin VI Domain: IMP dehydrogenase: Imp-1 Metallo Hypothetical 28.8 Kda Protein In Psd1-Sk: Hypothetical 32.1 Beta-Lactamase; Indole-3-Glycerol Phosphate Synthase: Kda Protein. In Adha-Rcal I: Hypothetical 65.0 Kda Protein Indole-3-Glycerol-Phosphate Synthase; Indole-3-Pyruvate In Cox 14-C: Hypothetical 65.0 Kda Protein In Cox 14-Cos 3: Decarboxylase; Indoleamine 2,3-dioxygenase; Inducible Hypothetical Abc Transporter ATP-Binding; Hypothetical Nitric Oxide Synthase; Influenza A Subtype N2 Neuramini Abc Transporter ATP-Binding Pro; Hypothetical Isochoris dase; Influenza A Subtype N9 Neuraminidase; Influenza matase Family Protein: Hypothetical Oxidoreductase Yahf: Virus B/Lee/40 Neuraminidase; Inorganic Polyphosphate/ Hypothetical Oxidoreductase Yiak; Hypothetical Oxi ATP-Glucomannokinase; Inorganic Polyphosphate/ATP doreductase Yghd: Hypothetical Protein: Hypothetical Pro Nad Kinase; Inorganic Pyrophosphatase; Inosine Monophos tein Af0103: Hypothetical Protein Af1403; Hypothetical Pro phate Dehydrogenase 2: Inosine Monophosphate tein Af1521: Hypothetical Protein Af1796: Hypothetical Dehydrogenase I: Inosine-5'-Monophosphate Dehydroge Protein Af2198: Hypothetical Protein Agr L 1239; Hypo nase; Inosine-5'-Monophosphate Dehydrogenase 2: Inosine thetical Protein Alriš027; Hypothetical Protein Apc35924; Adenosine-Guanosine Preferring Nucle; Inosine-Adenosine Hypothetical Protein Apc36103: Hypothetical Protein Guanosine-Preferring Nucle; Inositol 1,3,4-Trisphosphate Aq 328: Hypothetical Protein Bsu33890: Hypothetical Pro 5/6-Kinase; Inositol 1,4,5-Trisphosphate Receptor Type 1: tein EgcO68; Hypothetical Protein Fli11149; Hypothetical Inositol Monophosphatase; Inositol-3-Phosphate Synthase: Protein HiO828: Hypothetical Protein Hil388.1: Hypotheti Inositol-3-Phosphate Synthase; Inositol-Trisphosphate 3-Ki cal Protein Lech: Hypothetical Protein Mds018: Hypotheti nase A, Insecticyanin A Form: Insulin; Insulin receptor, Insu cal Protein M1247; Hypothetical Protein PaO094; Hypo lin-Like Growth Factor I; Insulin-Like Growth Factor Recep thetical Protein Pa2260: Hypothetical Protein Pa3270; tor 1: Intact Lactose Operon Repressor With Gratuit; Hypothetical Protein Pa3967: Hypothetical Protein Pa-Ho: Integrase; Integrin alpha-4: Integrin Alpha-L: Integrin Beta-4 Hypothetical Protein pH0236: Hypothetical Protein pH0642: Subunit; Intercellular Adhesion Molecule-1, Intercellular Hypothetical Protein pH1313; Hypothetical Protein pH1602; Adhesion Molecule-2; Interferon gamma receptor including Hypothetical Protein pH1897: Hypothetical Protein pH1917: IFNGR1 and IFNGR2 (IFN-gamma binds directly to Hypothetical Protein Rbstp0775: Hypothetical Protein IFNGR1 and indirectly to IFNGR2); Interferon receptor Rv0793; Hypothetical Protein Rv0819; Hypothetical Protein IFNAR1; Interferon receptor IFNAR2c; Interferon Stimu US 2015/0309021 A1 Oct. 29, 2015 50 lated Gene 20Kda; Interferon-Beta; Interferon-Inducible Reductase: L-Fucose Isomerase: L-Fuculose 1-Phosphate Gtpase: Interleukin 17F: Interleukin-1 type I receptor (IL-1 Aldolase; L-Fuculose-1-Phosphate Aldolase; L-Histidinol RI); Interleukin-11 receptor alpha chain (IL-11 R-alpha); Dehydrogenase; light chain; Light chain 1 B72.3 (murine); Interleukin-12 Beta Chain; Interleukin-19: Interleukin-2: Lignin Peroxidase; Limonene-1,2-Epoxide Hydrolase; Interleukin-2 receptor alpha chain (IL-2-RA); Interleukin-2 Lipase; Lipase; Lipase 2: Lipase 3: Lipase, Gastric: Lipid receptor beta chain (IL-2-RB); Interleukin-3 precursor; Inter Transfer Protein; Lipi; Lipoate-Protein Ligase, Putative: mediate conductance Ca(2+)-activated K(+) channel, hIK1; Lipoprotein Mxim; Lipoprotein Nlpi; Lipoxygenase-3; Internalin A: Interphotoreceptor retinoid-binding protein; Lipoyltransferase 1: Lithostathine: Liver Alcohol Dehydro Intestinal Fatty Acid-Binding Protein; Intramolecular Trans genase; Liver Carboxylesterase: Liver Carboxylesterase I; Sialidase; Intron-Associated Endonuclease 1: Invasin; Inver Liver Fatty Acid Binding Protein: Liver Glycogen Phospho tase Inhibitor; Inward rectifier potassium channel 2: Iols Pro rylase; L-Lactate Dehydrogenase; L-Lactate Dehydroge tein; Ionotropic Glutamate Receptor 5; Iota Toxin nase; L-lactate dehydrogenase A chain; L-lactate dehydroge Component Ia, Iota-Carrageenase; Iron; Iron Binding Protein nase A-like 6A: L-lactate dehydrogenase A-like 6B; L-lactate Fbpa; Iron-Utilization Periplasmic Protein; Isoasparty1 dehydrogenase B chain; L-Lactate Dehydrogenase B Chain; Dipeptidase; Isocitrate Dehydrogenase; Isocitrate Dehydro L-lactate dehydrogenase C chain; L-Lactate Dehydrogenase genase; Isocitrate dehydrogenase NAD subunit alpha, mito H Chain; L-Lactate Dehydrogenase M Chain; L-Lactate/ chondrial precursor: Isocitrate dehydrogenase NAD sub Malate Dehydrogenase; Lmaj004.091Aaa; Lmaj004144Aaa unit beta, mitochondrial precursor, Isocitrate dehydrogenase Protein; Long-Chain Fatty Acid Transport Protein; Long NAD Subunit gamma, mitochondrial precursor; Isocitrate chain-fatty-acid-CoA ligase 1: Long-chain-fatty-acid-CoA Dehydrogenase Nadip Cytoplasmic; Isocitrate Lyase; ligase 3; Long-chain-fatty-acid-CoA ligase 4: Long-Chain Isoflavone 0-Methyltransferase: Isolectin B4: Isoleucine Fatty-Acid-CoaSynthetase; Low density lipoproteins (LDL): tRNA synthetase; Isoleucyl-tRNA Synthetase; Isoliquiritige Low-affinity cationic amino acid transporter-2: Low-Density nin 2'-O-Methyltransferase: Isomerase: Isopenicillin N Syn Lipoprotein Receptor, Lc.2: L-Rhamnose Isomerase; thase; Isopentenyl Diphosphate Delta-Isomerase; L-serine dehydratase; L-Sulfolactate Dehydrogenase; Isopentenyl-Diphosphate Delta-Isomerase: Isovaleryl-Coa L-Threonine-O-3-Phosphate Decarboxylase; L-type amino Dehydrogenase; Ispa/Ispf Bifunctional Enzyme: Iswi Pro acid transporter 1 (LAT1); L-type amino acid transporter 2; tein; Kallikrein; Kallikrein 1: Kallikrein 6: Kanamycin nucle Luciferase; Lumazine Synthase; Luteinizing Hormone otidyltransferase; Kappa-4 Immunoglobulin; Kata Catalase; Releasing Hormone (LHRH) Receptor; L-Xylulose Reduc Kdo-8-Phosphate Synthetase; Kdpg Aldolase; Ketoacyl tase; Lymphocyte function-associated antigen 1 (CD11a anti Reductase; Kex1; Killer Cell Immunoglobulin-Like Recep gen); Lysine Biosynthesis Enzyme: Lysine hydroxylase; tor 2Ds; Kindling Fluorescent Protein; Kinesin; Kinesin Lysozyme: Lysozyme C; Lysozyme Insertion Mutant With Heavy Chain; Kinesin Heavy Chain-Like Protein; Kinesin Ala Inserted; Lysozyme: Lysozyme Mutant With Cys 54 Motor Ncd; Kinesin-Like Protein Kar3; Kinesin-Like Protein Replaced By Thr; Lysyl Oxidase; Lysyl Oxidase; Lysyl Kifl 1; Kinesin-Like Protein Kifl A: Kinesin-Like Protein tRNA synthetase; Lytic Murein Transglycosylase B; Kif2C; Kinesin-Related Motor Protein Eg5; Kynureninase; M=4=Lactate Dehydrogenase; M1 muscarinic acetylcholine Kynurenine/alpha-aminoadipate aminotransferase mito receptor; M2 muscarinic acetylcholine receptor; Mac-2 Bind chondrial; Kynurenine-Oxoglutarate Transaminase I, L., L-2- ing Protein; Macromomycin; Macrophage Metalloelastase; Haloacid Dehalogenase; L-2-Hydroxyisocaproate Dehydro Macrophage Migration Inhibitory Factor; Magnesium-De genase; L-3-Hydroxyacyl Coa Dehydrogenase; L-3- pendent Phosphatase-1; Major Allergen I Polypeptide, Fused Hydroxyacyl-Coa Dehydrogenase; L-3-Phosphoserine Chain 2: Major Autolysin; Major Capsid Protein; Major Phosphatase; Laccase; Laccase; Laccase 1: Laccase 2: Lac Envelope Protein E: Major Nad; Major Pollen Allergen BetV tadherin: Lactaldehyde Reductase; Lactate Dehydrogenase; 1-L; Major Urinary Protein; Major Urinary Protein 2: Major Lactate Dehydrogenase; Lactoferrin; Lactoferrin; Lactose Urinary Protein I; Malate Dehydrogenase; Malate Dehydro Permease; Lactotransferrin; Lactoylglutathione Lyase; genase; Malate dehydrogenase, cytoplasmic; Malate Dehy L-Alanine Dehydrogenase; L-Allo-Threonine Aldolase; drogenase, Glyoxysomal; Malate dehydrogenase, mitochon Lambda Exonuclease; Laminarinase 16A: L-Amino Acid drial precursor; Malate Synthase G. Male-B363; Malic Oxidase; Lanosterol Synthase; Lantibiotic Mersacidin; Enzyme: Malic Enzyme 2: Malonamidase E2: Malonyl Coa: L-Arabinose-Binding Protein: L-Arabinose-Binding Protein Acyl Carrier Protein Malonyltra; Maltodextrin Glycosyl Complex With L-A; Large T Antigen; L-Arginine: Glycine transferase: Maltodextrin Phosphorylase; Maltodextrin Amidinotransferase; L-Arginine\: Glycine Amidinotrans Binding Protein; Maltodextrin-Binding Protein Male-B133; ferase; L-Asparaginase; L-Asparagine Amidohydrolase; Maltogenic Amylase; Maltooligosyl Trehalose Synthase: L-Aspartate Ammonia-Lyase, L-Aspartate Oxidase; Lck Maltooligosyltrehalose Trehalohydrolase; Maltoporin; Mal Kinase; L-Cysteine/L-Cystine C-S Lyase; Lectin; Lectin; tose Binding Protein Fused With Designed; Maltose Trans Lectin Cel-I, N-Acetyl-D-Galactosamine-Speci; Lectin port Protein Malk; Maltose-6'-Phosphate Glucosidase; Mal Complex With Lactose; Lectin Pal; Lectin, Isoform 1: Lectin tose-Binding Periplasmic Protein; Maltose-Binding Protein; D2: Leghemoglobin: Leghemoglobin A; Lens Fiber Major Maltose-Binding Protein Mutant Male31; Maltotetraose Intrinsic Protein; Lethal; Lethal Factor; Leucine Aminopep Forming Amylase; Mandelate Racemase; Manganese Peroxi tidase; Leucine carboxyl methyltransferase 1: Leucine car dase; Manganese-Dependent Inorganic Pyrophosphatas; boxyl methyltransferase 2: Leucoagglutinating Phytohemag Mannan Endo-1,4-Beta-Mannosidase; Mannanase A. Man glutinin; Leucoanthocyanidin Dioxygenase; Leucyl-tRNA nitol Dehydrogenase; Mannose Receptor; Mannose-6-Phos Synthetase; Leucyl-tRNA synthetase, cytoplasmic; Leukoag phate Isomerase: Mannose-Binding Protein A: Mannose glutinin; Leukocidin F Subunit; Leukocyte Elastase; Leuko Binding Protein Associated Serin; Mannose-Binding Protein sialin; Leukotriene A-4 Hydrolase; Leukotriene B4 12-Hy C; Mannose-Binding Protein-A; Mannose-Binding Protein droxydehydrogenase/Pros; Levansucrase; Levodione C; Mannosyl-Oligosaccharide 1.2-Alpha-Mannosida; Man US 2015/0309021 A1 Oct. 29, 2015 nosyl-Oligosaccharide Alpha-1,2-Mannosida; Map Kinase boxylate transporter 1; Monocarboxylate transporter 2: P38; Map Kinase-Activated Protein Kinase 2: Maspin Pre Monocarboxylate transporter 3: Monocarboxylate trans cursor, Mast/stem cell growth factor receptor precursor, porter 4: Monocarboxylate transporter 5; Monocarboxylate Matrix Gla-protein; Matrix metalloprotease 2 (MMP-2); transporter 6: Monocarboxylate transporter 7: Monocarboxy matrix metalloprotease 9 (MMP-9); Matrix Metalloprotein late transporter 8: Monocyte Chemotactic Protein 2: Mono ase 3: Matrix Metalloproteinase-16; Matrix Metalloprotein cyte Differentiation Antigen Cd14; Mono-Heme C-Type ase-2: Matrix Metalloproteinase-8; Matrix Porin; Matrix Cytochrome Scya; Monomer Hemoglobin Component III; Porin Outer Membrane Protein F; Matrix protein M2: Mdc Monomer Hemoglobin Component IV: Monomeric Sar Sign1B Type I Isoform; Mdc-Sign2 Type I Isoform; Medium cosine Oxidase; Monomethylamine Methyltransferase Chain Acyl-Coa Dehydrogenase; Melatonin Receptor; Mela Mtmb1; Morphinone Reductase: Motuporin; M-Phase tonin receptor type 1B: Membrane Copper Amine Oxidase; Inducer Phosphatase 2: Mre 11 Nuclease; Mrna Capping Menb: Merozoite Surface Protein-1; Meso-Diaminopimelate Enzyme; Mrna Decapping Enzyme; Mrsd Protein; Mta/Sah D-Dehydrogenase; Metabotropic glutamate receptor 1; Nucleosidase; Mu Class Glutathione S-Transferase Of Metabotropic Glutamate Receptor Subtype 1; Meta-Cleav Isoenz; Mu Class Tetradeca-; Mu-Conotoxin Piiia; age Product Hydrolase; Metallo Beta-Lactamase II; Metal Mu-Conotoxin Smilia; Mucosal Addressin Cell Adhesion lochaperone Atx1; Methionine adenosyltransferase: Molecule-1; Mu-crystallin homolog; Multidrug Resistance Methionine Aminopeptidase; Methionine Aminopeptidase 2: Abc Transporter ATP-Bin: Multidrug Resistance Protein; Methionine Gamma-Lyase; Methionine Synthase; Methion Multidrug Resistance Protein Mexa; Multidrug-Efflux Trans ine synthase reductase; Methionine-R-sulfoxide reductase: porter 1 Regulator Bmr: Multiple. Antibiotic Resistance Pro Methionine-R-sulfoxide reductase B2: Methionyl Ami tein Marr; Multiple T-cell antigens (CD1, CD2, CD3, CD5, nopeptidase; Methionyl-tRNA synthetase; Methoxy Mycolic CD5, CD7. CD8); multivitamin transporter; Muscarinic ace Acid Synthase 2: Methuselah Ectodomain; Methyl-Accept tylcholine receptor M3; Muscarinic acetylcholine receptor ing Chemotaxis Protein; Methylaspartate Mutase S Chain; M4; Muscarinic acetylcholine receptor M5; Mutator Mutt Methylated-DNA-protein-cysteine methyltransferase: Meth Protein; Muth; Mycotic Acid Synthase: Myoglobin: Myohe ylcrotonoyl-CoA: Methylcrotonoyl-CoA 2; Methylene Tet merythrin: Myo-Inositol Hexaphosphate Phosphohydrol; rahydromethanopterin Dehydrogen; Methylenetetrahydro Myo-Inositol Hexaphosphate Phosphohydrolase; Myo folate Dehydrogenase/Cy; Methylenetetrahydrofolate Inositol Monophosphatase: Myo-Inositol-1-Phosphate Syn reductase; Methylglyoxal Synthase; Methylmalonate-semi thase: Myo-Inositol-1-Phosphate Synthase; Myo-Inositol-1- aldehyde dehydrogenase; Methylmalonic aciduria protein; Phosphate Synthase-Related Pr: Myosin: Myosin le Heavy Methylmalonyl Coa Decarboxylase; Methylmalonyl-Coa Chain; Myosin II Heavy Chain; Myosin II Heavy Chain Carboxyltransferase 12S Su: Methylmalonyl-CoA mutase, Fused To Alpha-Actini; Myosin-3 Isoform; Myristoyl-Coa: mitochondrial precursor: Mevalonate Kinase; Mgp-40; Mhc Protein N-Myristoyltransferas; Myrosinase; N Utilization Class I Homolog Mic-A; Microcystin-Lr: microsomal trig Substance Protein B Homolog: N,N-Dimethylglycine Oxi lyceride transfer protein; Microtubule Motor Protein Ncd; dase; N-4 Cytosine-Specific Methyltransferase Pvu; N5-Car Mimochrome IV. Miniaturized Metalloprotei; Mineralocor boxyaminoimidazole Ribonucleotide Synt; Na+/K+/2C1-co ticoid Receptor; Minor Core Protein Lambda 3: Mitchondrial transporter, N-Acetylgalactosamine-4-Sulfatase; carnitine/acylcarnitine carrier protein CACL; Mitochondrial N-Acetylglucosamine-1-Phosphate Uridyltransf: N-Acetyl Aconitase; Mitochondrial Aldehyde Dehydrogenase; Mito glucosamine-6-Phosphate Deacetylase; N-Acetyllac chondrial aspartate-glutamate carrier protein; Mitochondrial tosaminide Alpha-1,3-Galactosylt; N-Acetylactosaminide carnitine/acylcarnitine carrier protein; Mitochondrial folate Alpha-1,3-Galactosyltr; N-Acetyl-L-Ornithine Carbamoyl transporter/carrier; Mitochondrial glutamate carrier 1; Mito transferase; N-Acetylneuraminate Lyase; N-Acetylneurami chondrial glutamate carrier 2: Mitochondrial ornithine trans nate Lyase Subunit; N-Acylamino Acid Racemase: N-acyl porter 1; Mitochondrial ornithine transporter 2: Mitogen glucosamine 2-epimerase; Nad; NAD(P) transhydrogenase, Activated Protein Kinase 10; Mitogen-Activated Protein mitochondrial precursor; Nad-Dependent Alcohol Dehydro Kinase 14; Moad Related Protein; Modification Methylase genase; Nad-Dependent Deacetylase 2; Nad-Dependent For Hhai; Modification Methylase Rsri; Module-Substituted Chi mate Dehydrogenase; Nad-Dependent Malic Enzyme: Nad mera Hemoglobin Beta-A; Mol Id: 1; Molecule: 6-Phospho Dependent Malic Enzyme: Nad-Dependent Malic Enzyme, Beta-Glucosid; Mol Id: 1; Molecule: Adenylate Kinase; Mitochondria; NAD-dependent malic enzyme, mitochon Chain; Mol Id: 1; Molecule: Bacteriorhodopsin; Mol Id: 1; drial precursor; Nadh Oxidase; Nadh Oxidase/Nitrite Reduc Molecule: Beta-Lactamase II: Syno; Mol Id: 1; Molecule: tase; Nadh Peroxidase; Nadh Pyrophosphatase; Nadh-Azore Carbamate Kinase-Like C; Mol Id: 1; Molecule: Cyto ductase, Fmn-Dependent: NADH-cytochrome b5 reductase: chrome C2; Chain: A Mol Id: 1; Molecule: Endo-1,4-Beta Nadh-Dependent Butanol Dehydrogenase; NADH Xylanase; Mol Id: 1; Molecule: Glutaredoxin; Engineered; ubiquinone oxidoreductase 13 kDa-A subunit, mitochondrial Mol Id: 1; Molecule: Maltodextrin Phosphoryl; Mol Id: 1: precursor, NADH-ubiquinone oxidoreductase 13 kDa-B sub Molecule: Progesterone Receptor; Molecule: Apomyoglo unit; NADH-ubiquinone oxidoreductase 15 kDa subunit; bin; Other Details: Cryst; Molecule: Cathepsin B; Ec: 3.4. NADH-ubiquinone oxidoreductase 18 kDa subunit, mito 22.1; Mutatio; Molecule: Human ADP-Ribosylation Factor chondrial precursor, NADH-ubiquinone oxidoreductase 19 1; S: Molecule: Protein Kinase C Delta Type; Domain; kDa subunit; NADH-ubiquinone oxidoreductase 20 kDa sub Molybdenum Cofactor Biosynthesis Protein; Molybdenum unit, mitochondrial precursor, NADH-ubiquinone oxi Cofactor Biosynthesis Protein A; Molybdopterin Biosynthe doreductase 23 kDa subunit, mitochondrial precursor: sis Cnx1; Molybdopterin Converting Factor Subunit 2: NADH-ubiquinone oxidoreductase 24 kDa subunit, mito Molybdopterin-Guanine Dinucleotide Biosynthe: Momor chondrial precursor, NADH-ubiquinone oxidoreductase 30 din; Mono-ADP-Ribosyltransferase C3; Monoamine oxidase kDa subunit, mitochondrial precursor, NADH-ubiquinone A (MAO-A); Monoamine oxidase B (MAO-B); Monocar oxidoreductase 39 kDa subunit, mitochondrial precursor: US 2015/0309021 A1 Oct. 29, 2015 52

NADH-ubiquinone oxidoreductase 42 kDa subunit, mito Transhydrogenase; Nicotinamide Nucleotide Transhydroge chondrial precursor, NADH-ubiquinone oxidoreductase 49 nase, Su; Nicotinamide-Nucleotide Adenylyltransferase: kDa subunit, mitochondrial precursor, NADH-ubiquinone Nicotinate D-ribonucleotide phyrophsopate phosphoribosyl oxidoreductase 51 kDa subunit, mitochondrial precursor: transferase: Nicotinate Mononucleotide:5,6-Dimethylbenzi; NADH-ubiquinone oxidoreductase 75 kDa subunit, mito Nicotinate N-methyltransferase: Nicotinate Phosphoribosyl chondrial precursor, NADH-ubiquinone oxidoreductase 9 transferase: Nicotinate Phosphoribosyltransferase From Th; kDa subunit, mitochondrial precursor, NADH-ubiquinone Nicotinate-Nucleotide Adenylyltransferase; Nicotinate oxidoreductase AGGG subunit, mitochondrial precursor: Nucleotide-Dimethylbenzimidazole; nicotinic acetylcholine NADH-ubiquinone oxidoreductase ASHI subunit, mitochon (ganglion) receptor, Nicotinic acetylcholine Receptor drial precursor, NADH-ubiquinone oxidoreductase B12 sub alpha2/alpha3; Nima-Related Protein; Nine-Haem Cyto unit; NADH-ubiquinone oxidoreductase B14 subunit; chrome C; Nine-Heme Cytochrome C; Nit-Fragile Histidine NADH-ubiquinone oxidoreductase B15 subunit; NADH Triad Fusion Protein; Nitrate Reductase; Nitric Oxide Reduc ubiquinone oxidoreductase B16.6 subunit; NADH tase; Nitric Oxide Synthase: Nitric oxide synthase IIB: Nitric ubiquinone oxidoreductase B17 subunit; NADH-ubiquinone Oxide Synthase, Inducible; Nitric-Oxide Reductase Cyto oxidoreductase B18 subunit; NADH-ubiquinone oxi chrome P450 55A1: Nitric-Oxide Synthase: Nitric-oxide doreductase B22 subunit; NADH-ubiquinone oxidoreductase synthase brain; Nitric-Oxide Synthase Homolog; Nitric-ox B8 subunit; NADH-ubiquinone oxidoreductase B9 subunit; ide synthase IIC; Nitric-Oxide Synthase, Brain; Nitric-Oxide NADH-ubiquinone oxidoreductase chain 1: NADH Synthase, Endothelial; Nitrite Reductase: Nitrogen Fixation ubiquinone oxidoreductase chain 2: NADH-ubiquinone oxi Regulatory Protein Fixl; Nitrogen Regulation Protein; Nitro doreductase chain 3: NADH-ubiquinone oxidoreductase gen Regulatory Iia Protein; Nitrogen Regulatory Protein Pii; chain 4: NADH-ubiquinone oxidoreductase chain 4L: Nitrogenase Iron Protein; Nitrogenase Iron Protein 1: Nitro NADH-ubiquinone oxidoreductase chain 5; NADH phorin 1: Nitrophorin 2: Nitrophorin 4: Nitroreductase: ubiquinone oxidoreductase chain 6: NADH-ubiquinone oxi Nitroreductase Family Protein; Nitrosocyanin; Nitrous Oxide doreductase KFYI subunit, mitochondrial precursor, NADH Reductase: Nitrous-Oxide Reductase; Nk Receptor; NMDA ubiquinone oxidoreductase MLRQ subunit: NADH receptor; N-Methyl-D-Aspartate Receptor Subunit 1: Nimra; ubiquinone oxidoreductase MLRQ subunit homolog: Nogalonic Acid Methyl Ester Cyclase. Non Catalytic Protein NADH-ubiquinone oxidoreductase MNLL subunit; NADH 1: Nonaheme Cytochrome C. Non-ATP Dependent L-Selec ubiquinone oxidoreductase MWFE subunit: NADH tive Hydantoina; Non-Catalytic Protein 1: Non-heme chlo ubiquinone oxidoreductase PDSW subunit: NADH roperoxidase; Nonspecific Lipid Transfer Protein: Nonspe ubiquinone oxidoreductase SGDH subunit, mitochondrial cific Lipid-Transfer Protein: Nonstructural Polyprotein precursor, NADH-ubiquinone oxidoreductase subunit B14. Pp 1A: Non-Symbiotic Hemoglobin: Not Applicable: 5a; NADH-ubiquinone oxidoreductase subunit B14.5b: NPC1L1 (Niemann-Pick C1 Like 1) Protein transporter; NADH-ubiquinone oxidoreductase subunit B14.7; NADH Npqtn Specific Sortase B: Nrdi Protein; NRH dehydrogenase ubiquinone oxidoreductase subunit B17.2: Nadp: Nadp quinone 2: Nitpase P4: Nuclear Inclusion Protein A; Nuclear Dependent Non Phosphorylating Glycerald; Nadp-Depen receptor OB 1: Nuclear Receptor Ror-Alpha; Nucleoside dent Alcohol Dehydrogenase; NADP-dependent malic 2-Deoxyribosyltransferase: Nucleoside Diphosphate Kinase; enzyme; NADP-dependent malic enzyme, mitochondrial Nucleoside Diphosphate Kinase A. Nucleoside Diphosphate precursor; Nadp-Dependent Mannitol Dehydrogenase; Kinase II: Nucleoside Diphosphate Kinase, Cytosolic; Nadp-Dependent Nonphosphorylating Glyceralde; Nadph Nucleoside Diphosphate Transferase: Nucleoside-Specific Dehydrogenase 1: Nadph Dependent Thioredoxin Reduc Channel-Forming Protein; Nudix Homolog; 06-Alkylgua tase; Nadph\: Ferredoxin Oxidoreductase; Nadph-Cyto nine-DNA Alkyltransferase: Obelin; Odorant Binding Pro chrome P450 Reductase; Nadph-Flavin Oxidoreductase: tein; Odorant Binding Protein Lush; Odorant-Binding Pro Nadp-Malate Dehydrogenase; Nagd Protein, Putative: Naip: tein; Old Yellow Enzyme: Olfactory Marker Protein: Namn Adenylyltransferase; Nbla; N-Carbamyl-D-Amino Oligopeptide Abc Transporter, Periplasmi; Omegac-Txix: Acid Amidohydrolase; Ndx1; Nei Endonuclease Viii-Like 1: Omp Synthase; Ompk36; Opioid delta Receptor (OP1); Neocarzinostatin; Neopullulanase; Neopullulanase 2: Opioid kappa Receptor (OP2); Opioid mu Receptor (OP3); Neprilysin; N-Ethylmaleimide Sensitive Fusion Protein; Opioid sigma 1 Receptor; Opioid sigma Receptor (OP4); Neural Globin: Neural Hemoglobin; Neural Kinase, Orange Carotenoid Protein; Orc2: Orexin-A.; Orf, Conserved Nuk=Eph/Elk/Eck Family Recept; Neuraminidase; Hypothetical Protein; Orf3; Organic acid transporter 3 Neuraminidase; Neuraminidase N2; Neuraminidase N9; (OAT3); organic anion transporter (MRP1, MRP2 and Neuroglobin; Neuronal Calcium Sensor 1: Neuropeptide Y: MRP3, MRP4 and MRP5); Organic cation/carnitine trans Neuropsin; Neurotensin receptor type 2: Neurotoxin Bmk porter 1; Organic cation/carnitine transporter 2: Organic M4; Neurotoxin Bmk37; Neutral amino acid transporter A: Hydroperoxide Resistance Protein: Ornithine Aminotrans Neutral amino acid transporter B: m Neutral Protease II; ferase: Ornithineaminotransferase, mitochondrial; Ornithine Neutrophil Collagenase; Neutrophil elastase; Neutrophil Carbamoyltransferase: Ornithine carbamoyltransferase, Gelatinase; Neutrophil Gelatinase-Associated Lipocalin; mitochondrial; Ornithine Cyclodeaminase; Ornithine Decar Neutrophil-Activating Protein A; NG.NG-dimethylarginine boxylase: Ornithine decarboxylase antizyme: Ornithine dimethylaminohydrolase 1: NG.NG-dimethylarginine dim Transcarbamoylase; Orotidine 5'-Monophosphate Decar ethylaminohydrolase 2: Nh; Niacin receptor HM74A: Nickel boxylase: Orotidine 5'-Phosphate Decarboxylase; Orotidine Responsive Regulator; Nicotinamide Mononucleotide Ade Monophosphate Decarboxylase; Orphan Nuclear Receptor nylyl Tra; Nicotinamide Mononucleotide Adenylyl Transfe; PXr: Osmolarity Sensor Protein; Osmoprotection Protein; Nicotinamide Mononucleotide Adenylyltransfer; Nicotina Osmotically Inducible Protein C; Osteocalcin; O-Succinyl mide Mononucleotide Adenylyltransfer; Nicotinamide benzoate Synthase: Outer Membrane Lipoprotein Lolb: Nucleotide Transhydrogenase; Nicotinamide Nucleotide Outer Membrane Phospholipase A: Outer Membrane Protein US 2015/0309021 A1 Oct. 29, 2015

A: Outer Membrane Protein F: Outer Membrane Protein Lysozyme Insertion Mutant With Ser; Phaseolin; Phenazine Nspa: Outer Membrane Protein Tolc; Outer Membrane Pro Biosynthesis Protein Phzd; Phenazine Biosynthesis Protein tein X: Outer Protein Yopm: Ovalbumin; Ovocleidin; Ovo Phzf: Phenazine Biosynthesis Protein Phzg; Phenol 2-Hy mucoid: Ovotransferrin; Ovotransferrin; Oxygen-Insensitive droxylase Component B: Phenol 2-Monooxygenase; Phenol Nad: Oxygen-insensitive NAD(P)H nitroreductase; Oxygen Hydroxylase; Phenylacetone Monooxygenase; Phenylala Insensitive Nadph Nitroreductase; Oxysterols Receptor LXr nine Ammonia-Lyase; Phenylalanine Ammonia-Lyase 1: Beta: Oxytocin receptor (Neurophysin 1); P protein In Phenylalanine Hydroxylase; Phenylalanine-4-Hydroxylase; cludes: DNA-directed DNA polymerase: P protein Includes: Phenylalanine-Regulated 3-Deoxy-D-Arabino-H; Phenyla DNA-directed DNA polymerase, RNA-directed DNA poly lanyl-tRNA synthetase; Phenylalanyl-tRNA synthetase alpha merase, Ribonuclease H; P/neurokinin 1 Receptor; P1 chain; Phenylalanyl-tRNA synthetase beta chain; Phenyle Nuclease; P2 Myelin Protein; P2 Protein; P2Y12 platelet thanolamine N-Methyltransferase; Phenylethylamine Oxi ADP Receptor; P-30 Protein; P300/Cbp Associating Factor; dase; Pheromone Binding Protein; Pheromone-Binding Pro P35; P450 Epoxidase; P450 Monooxygenase; P450Cin: tein; Pheromone-Binding Protein Asp1; Phoq Histidine P64K; P97; Palmitoyl Protein Thioesterase 1: Palmitoyl-Pro Kinase; Phosphatase; Phosphatase; Phosphate Acetyltrans tein Thioesterase 2 Precursor P-Aminobenzoate Synthase ferase; Phosphate-Binding Protein: Phosphatidylcholine Component I: pancreatic alpha amylase; Pancreatic alpha Transfer Protein: Phosphatidylethanolamine Binding Pro amylase precursor, Pancreatic Hormone; Pancreatic Lipase tein: Phosphatidylethanolamine-Binding Protein: Phosphati Related Protein 1: Pancreatic Lipase Related Protein 2; pan dylinositol 3-Kinase Catalytic Subu: Phosphatidylinositol creatic triacylglycerol lipase; Pancreatic triacylglycerol Phosphate Phosphatase; Phosphatidylinositol Transfer Pro lipase Precursor; Pancreatic Trypsin Inhibitor; Pantoate tein A1: Phosphatidylinositol Transfer Protein Alpha; Phos Beta-Alanine Ligase; Pantoate-Beta-Alanine Ligase; Pan phatidylinositol Transfer Protein Sec14P: Phosphatidylinosi tothenate kinase; Pantothenate Synthetase; Papain; Para-ami tol-Specific Phospholipase: Phosphatidylserine nobenzoate synthase component I; Para-aminobenzoic acid decarboxylase proenzyme; Phosphatidylserine receptor; (PABA); Parathion Hydrolase; Parm; Parvalbumin; Parvalbu Phospho-2-Dehydro-3-Deoxyheptonate Aldolase; Phospho min Alpha; Pbcv-1 DNA Ligase; Pcza361.16; P-D-Man 2-Dehydro-3-Deoxyheptonate Aldolase; Phosphocarrier nopyranosyl Ester), GDP-Mannose 6-Dehydrogenase; Pea Protein Hpr: Phosphodiesterase 4D: Phosphoenolpyruvate Lectin: Peanut Lectin: Peanut Peroxidase, Major Cationic Carboxykinase; Phosphoenolpyruvate Carboxykinase, Isozym; Pectate Lyase; Pectin Methylesterase; Penicillin V Cytos; Phosphoenolpyruvate Carboxykinase, Cytosolic; Acylase; Penicillin-binding protein 1A: Penicillin-binding Phosphoenolpyruvate Carboxylase; Phosphoenolpyruvate protein 1B: Penicillin-binding protein 2: Penicillin-Binding Mutase; Phosphoenolpyruvate Phosphomutase; Phosphoen Protein 2A; Penicillin-binding protein 2B: Penicillin-Bind Zyme Intermediate Of Fru-2,6-Bi; Phosphofructokinase; ing Protein 2X; Penicillin-binding protein 3: Penicillin-bind Phosphoglucose Isomerase; Phosphoglycerate kinase; Phos ing protein 4 precursor; Penicillin-Binding Protein 5: Peni phoglycerate kinase 1: Phosphoglycerate kinase, glycoso cillin-binding protein 5 precursor; Penicillin-binding mal; Phosphoglycerate Mutase; Phosphoglycerate Mutase 1: proteins 1A/1B: Penicillin-Insensitive Murein Endopepti Phosphoglycolate Phosphatase; Phosphoheptose Isomerase: dase; Penicillopepsin; Pentaerythritol Tetranitrate Reductase: Phosphoinositide-Specific Phospholipase C, I; Phospholi Penton Protein; Pentosyltransferase: Pepp 1: Peptaibol; Pep pase A=2=; Phospholipase A2; Phospholipase A2 Homolog: tide; Peptide Amidase; Peptide Deformylase; Peptide Phospholipase A2 Homolog 2: Phospholipase A2 inhibitory Deformylase 2: Peptide Deformylase Defb; Peptide protein; Phospholipase A2 Isoform 2: Phospholipase A2 Iso Deformylase Pdfl; Peptide Methionine Sulfoxide Reductase: form 3: Phospholipase A2, Major Isoenzyme: Phospholipase Peptide N-Myristoyltransferase: Peptide Transporter Tap1; C; Phospholipase C Delta-1: Phosphomannomutase; Phos Peptide-N: Peptidic Toxin Nodularin: Peptidoglycan Recog phomethylpyrimidine Kinase; Phosphonoacetaldehyde nition Protein I-Alph: Peptidoglycan Recognition Protein Sa Hydrolase; Phosphonoacetate Hydrolase; Phosphopanteth Cg 11709; Peptidoglycan synthetase fisl. Peptidyl-Glycine eine Adenylyltransferase: Phosphoribosyl Anthranilate Alpha-Amidating Monooxygena; Peptidylglycine Alpha Isomerase: Phosphoribosyl Pyrophosphate Synthetase; Phos Hydroxylating Monooxyg: Peptidyl-Lys Metalloendopepti phoribosylamidoimidazole-Succinocar; Phosphoribosy dase; Peptidyl-Prolyl Cis-Trans Isomerase; Peptidyl-Prolyl lamine-Glycine Ligase; Phosphoribosylformylglycina Cis-Trans Isomerase 5: Peptidyl-Prolyl Cis-Trans Isomerase midine Synth; Phosphoribosyiglycinamide A: Pepv: Peridinin-Chlorophyll Protein: Peripheral nerve Formyltransferase; Phosphoribosyltransferase-Related Pro Sodium channel 3: Peripheral nerve sodium channel 5; tein: Phosphorylated Map Kinase P38-Gamma; Phospho Peripheral Plasma Membrane Cask: Periplasmic Divalent serine Aminotransferase; Phosphoserine Phosphatase; Phos Cation Tolerance Pr; Periplasmic Divalent Cation Tolerance photriesterase: Phosphotriesterase Homology Protein; Protei: Periplasmic Molybdate-Binding Protein: Peroxidase; Photoactive Yellow Protein: Photolyase; Phy3 Protein: P-Hy Peroxidase C1A: Peroxidase N; Peroxidase/Catalase T: Per droxybenzoate Hydroxylase; P-Hydroxybenzoate Hydroxy oxiredoxin; Peroxiredoxin 5 Residues 54-214; Peroxiredoxin lase Complexed With; P-Hydroxybenzoate Hydroxylase 5, mitochondrial precursor; Peroxisomal bifunctional Mutant With Ty; Phytase; Phytochrome Response Regulator enzyme; Peroxisomal Carnitine O-Octanoyltransfer; Peroxi Rcpa; Phytochrome Response Regulator Rcpb; Phytohemag somal Carnitine O-Octanoyltransferase; Peroxisomal glutinin-L; Pi Glutathione Transferase: Pigment Epithelium Hydratase-Dehydrogenase-Epim; Peroxisomal Hydratase Derived Factor; Pituitary Adenylate Cyclase Activating Po: Dehydrogenase-Epimeras; Peroxisomal multifunctional Placenta Growth Factor; Plasma retinol-binding protein; enzyme type 2: Peroxisomal Trans 2-Enoyl Coa Reductase: Plasmepsin 2; Plasmepsin II: Plasminogen; Plasminogen Peroxisome Proliferator Activated Receptor A: Peroxisome Activator; Plasminogen Activator Inhibitor-1: Plasminogen Proliferator Activated Receptor D: Peroxisome Proliferator precursor; Plasmodium proton pump; Platelet Glycoprotein Activated Receptor G; pH 2.5 Acid Phosphatase; Phage T4 Ib Alpha Chain Precurs; Platelet-Activating Factor Acetylhy US 2015/0309021 A1 Oct. 29, 2015 54 drolas; Platelet-Activating Factor Acetylhydrolase; PML thetase; Prolyl Endopeptidase; Prolyl hydroxylase; Prolyl RAR Alpha Protein; Pms 1 Protein Homolog 2: Pnp Oxidase; Oligopeptidase; Prophospholipase A=2=; Propionyl-CoA Pokeweed Antiviral Protein; Pol Polyprotein; Pollen Allergen carboxylase alpha chain; Propionyl-CoA carboxylase beta Phl P1: Poly; Poly ADP-Ribose Polymerase-1; Polyamine chain; Propionyl-Coa Carboxylase Complex B Subunit; Oxidase; Polyandrocarpa Lectin; Polygalacturonase; Polyga Prostacyclin synthase; Prostaglandin D2 receptor; Prostag lacturonase I; Polygalacturonase II; Polygalacturonase Inhib landin E2 receptor, EP1 subtype: Prostaglandin E2 receptor, iting Protein; Polynucleotide Kinase; Polyomavirus Coat EP2 subtype: Prostaglandin Endoperoxide H Synthase-1; Protein Vp1; Polyomavirus Vp1 Pentamer; Polypeptide Prostaglandin F Synthase; Prostaglandin F2-alpha receptor; Deformylase; Polypeptide N-Acetylgalactosaminyltrans Prostaglandin G/HSynthase 1 Precursor: Prostaglandin G/H feras; Polyprotein; Polysialic Acid Capsule Biosynthesis Pro Synthase 2: Prostaglandin H2 Synthase: Prostaglandin H2 tein; Porcine Alpha-Amylase; Porcine Pancreatic Spas Synthase-1; Prostaglandin H2 Synthase-2: Prostaglandin molytic Polypepti; Porin; Porphobilinogen Deaminase; Receptor; Prostaglandin-E2 9-Reductase; Prostate-specific Porphobilinogen Synthase; Possible 3-Mercaptopyruvate membrane antigen (7E11-05.3 antigen/FOLH1-human/ Sulfurtransf: Possible G-T Mismatches Repair Enzyme: glutamate carboxypeptidase II); Prostatic Acid Phosphatase; Postsynaptic Density Protein; Postsynaptic Density Protein Protease; Protease II; Protease Retropepsin; Protease Syn 95; Potassium Channel; Potassium Channel Blocking Toxin thase and Sporulation Negati; Protease Vii; Protegrin 3: Pro 1; Potassium Channel Protein Rck4; Potassium channel sub tein: Ketosteroid Isomerase; Protein 1 D10; Protein Arginine family K member 1: Potassium channel subfamily K member Methyltransferase Prmt3; Protein Arginine N-Methyltrans 2: Potassium channel subfamily K member 6: Potassium ferase 1: Protein C: Protein Fkbi; Protein Kinase C. Protein large conductance calcium-activated channel. Subfamily M. Kinase C Interacting Protein; Protein Kinase C, Theta Type: alpha member 1: Potassium transporter (bacterial); Potas Protein Kinase Ck2: Protein Mat Protein Methyltransferase sium Voltage-gated channel Subfamily A member 1; Potas Hemk: Protein Ninb: Protein Rdmb: Protein With Similarity sium Voltage-gated channel Subfamily E member 1; Potas To Flavin-Contai; Protein Ybgc: Protein Ybhb: Protein Ycei: sium Voltage-gated channel Subfamily H member 2; Protein Yebr; Protein Yesu: Protein Yfbg: Protein Yof Pro Potassium voltage-gated channel subfamily H member 6: tein Ytnj; Protein: Igfbp-1Antagonist; Proteinase; Proteinase Potassium voltage-gated channel subfamily H member 7; K; Protein-Glutamine Gamma-Glutamyltransferase: Protein Potassium voltage-gated channel subfamily KQT member 1: Glutamine Glutamyltransferase E.; Protein-glutamine Potassium/Sodium Hyperpolarization-Activ; Potassium/So glutamyltransferase E precursor: Protein-Glutamine dium Hyperpolarization-Activated: Potassium-Transporting Glutamyltransferase E3: Protein-L-Isoaspartate; Protein-L- Atpase B Chain; Potassium-transporting ATPase beta chain; Isoaspartate O-Methyltransferase: Protein-Tyrosine Phos Pp60V-Src Tyrosine Kinase Transforming Prot; PPAR-alpha; phatase 1B: Protein-Tyrosine Phosphatase Yoph: Protein-Ty Ppc Decarboxylase Athal3A; Ppca; Ppr; Pr3; Precorrin-6Y rosine Phosphatase, Non-Recept; Protein-Tyrosine Methyltransferase/Putative: Precorrin-8X Methylmutase: Phosphatase, Non-Receptor T. Protein-Tyrosine-Phos Precursor Form Of Glucose-Fructose Oxidoredu: Precursor phatase; Prothrombin; Prothrombin; Prothrombin Fragment OfPeriplasmic Sugar Receptor; Predicted Amidotransferase: 1; Protoheme Ferrolyase; Proton-coupled amino acid trans Predicted Cobalamin Binding Protein; Preprotein Translo porter 1: Proto-Oncogene Serine/Threonine-Protein; Proto case Seca; Preprotein Translocase Seca 1 Subunit; Preprotein Oncogene Serine/Threonine-Protein Kina; Proto-Oncogene Translocase Seca Subunit; Prfa; Prion Protein; Probable Tyrosine-Protein Kinase Abl1; Proto-Oncogene Tyrosine 2-Phosphosulfolactate Phosphata; Probable Ammonium Protein Kinase Lck; Proto-Oncogene Tyrosine-Protein Transporter; Probable arabinosyltransferase A: Probable ara Kinase S. Proto-Oncogene Tyrosine-Protein Kinase Src.: Pro binosyltransferase B; Probable arabinosyltransferase C: tooncoprotein; Protoporphyrinogen Oxidase, Mitochondria; Probable Aromatic Acid Decarboxylase; Probable ATP-De P-Selectin; PseudoaZurin; Pseudocatalase; Pseudomonas pendent RNA Helicase P47: Probable Butyrate Kinase 2: Aeruginosa Lectin II: Psychrophilic Phosphatase I; Pteridine Probable Cell Division Inhibitor Mind: Probable Cysteine Reductase; Pteridine Reductase 1; Pteridine Reductase 2; Pts Desulfurase: Probable Fosfomycin Resistance Protein; Prob System, Chitobiose-Specific lib Comp; Pulmonary Surfac able Glutaminase Ybas; Probable Glutaminase Ybg; Prob tant-Associated Protein A: Pulmonary Surfactant-Associated able Gitpase Engc: Probable GTP-Binding Protein Enga: Protein D; Pumilio 1: Pur Operon Repressor; Pure: Purine Probable Inorganic Polyphosphate/ATP-Nad Kin: Probable Nucleoside Phosphorylase; Purine Regulatory Protein Yabi: Inorganic Polyphosphate/ATP-Nad Kin; Probable leucyl Purine Trans Deoxyribosylase; Purine-Nucleoside Phospho tRNA synthetase, mitochondrial; Probable low molecular rylase; Purple Acid Phosphatase; Purple Acid Phosphatase: weight protein-tyrosine-phosphatase epsP: Probable Malate Putative Acyl-Coa Thioester Hydrolase Hi0827; Putative Synthase G. Probable Methylisocitrate Lyase; Probable Alkylsulfatase Atsk; Putative Alpha-L-Fucosidase; Putative Polysaccharide Deacetylase Pdaa; Probable Pyridoxamine Aspartate Aminotransferase; Putative ATP-Dependent Clp 5'-Phosphate Oxidase; Probable S-Adenosylmethionine:2- Protease Proteoly; Putative Betaine Aldehyde Dehydroge Demethylmen; Probable Serine/Threonine-Protein Kinase; nase; Putative Blue Light Receptor; Putative Cellulase; Puta Probable Serine/Threonine-Protein Kinase Pel; Probable tive Cellulase Ce16: Putative Clc Family, Chlorine Transport Serine/Threonine-Protein Kinase Pkn; Probable tautomerase Prot: Putative Cytochrome P450; Putative Cytochrome P450 ydcE: Probable tyrosyl-tRNA synthetase, mitochondrial; 154A1: Putative Family 31 Glucosidase Yici; Putative Flavin Probable Uracil Phosphoribosyltransferase: Procarboxypep Oxidoreducatase; Putative Glur0 Ligand Binding Core: Puta tidase B; Proclavaminate Amidino Hydrolase; Profilin II; tive GlurO Ligand Binding Core: Putative G-protein coupled Progesterone Receptor; Progesterone Receptor (PR); Pro receptor 40; Putative Ketoacyl Reductase; Putative Lipase gramed Cell Death Protein 8: Programmed Cell Death Pro From The G-D-S-L Family: Putative Mannosyl-3-Phospho tein 8: Prolactin receptor precursor; Proline Dehydrogenase; glycerate Phospha; Putative Modulator Of DNA Gyrase: Proline Iminopeptidase; Proline oxidase; Proline-tRNA Syn Putative Nadph Dependent Oxidoreductases; Putative US 2015/0309021 A1 Oct. 29, 2015

Oxalate Decarboxylase; Putative Oxidoreductase RV2002: Rhamnogalacturonase A. Rhamnulose-1-Phosphate Aldo Putative Oxidoreductase RV2002: Putative Phosphatase: lase; Rhizome Secoisolariciresinol Dehydrogenase; Rhoa; Putative Polyprotein/Phosphatase; Putative Protease La Rhodanese; Rhodopsin; Rho-Related GTP-Binding Protein Homolog: Putative Riboflavin Kinase; Putative Snrnp Sm Rhoe: Riboflavin kinase; Riboflavin Kinase/Fmn Adenylyl Like Protein: Putative Sugar Kinase; Putative Transcriptional transferase: Riboflavin Synthase; Riboflavin Synthase Alpha Regulator; Putative Xylanase; Putidaredoxin Reductase: Chain; Ribokinase; Ribonuclease; Ribonuclease 1: Ribonu Putrescine-Binding Protein; Pyelonephritic Adhesin; Pyra clease 4: Ribonuclease A. Ribonuclease Hii; Ribonuclease nose Oxidase; Pyridoxal kinase; Pyridoxal Phosphate Bio Mc; Ribonuclease Mc1; Ribonuclease Pancreatic; Ribonu synthetic Protein Pdx; Pyridoxal phosphate phosphatase: clease pH; Ribonuclease Sa; Ribonuclease T1: Ribonuclease Pyridoxamine Kinase; Pyridoxine 5'-Phosphate Oxidase; U2: Ribonuclease UK114; Ribonuclease Z: Ribonuclease, Pyridoxine 5'-Phosphate Oxidase; Pyridoxine 5'-Phosphate Seminal; Ribonucleoside-Diphosphate Reductase 2 Alpha; Synthase; Pyridoxine-5'-phosphate oxidase; Pyrimidine Ribonucleoside-Diphosphate Reductase M2 Chai; Ribo Nucleoside Phosphorylase; Pyrogenic Exotoxin B: Pyro nucleotide reductase; Ribonucleotide Reductase R2: Ribo phosphatase: Pyrr Bifunctional Protein; Pyrroline-5-car nucleotide Reductase R2-2 Small Subunit; Ribonucleotide boxylate reductase 1: Pyrroline-5-carboxylate reductase 2: Reductase Subunit R2F: Ribose 5-Phosphate Isomerase: Pyrroline-5-carboxylate synthetase; Pyruvate carboxylase; Ribose-5-Phosphate Isomerase A: Ribose-5-Phosphate Pyruvate Decarboxylase; Pyruvate dehydrogenase; Pyruvate Isomerase Rpib: Ribosomal Protein L1; Ribosomal Protein Dehydrogenase E1 Component; Pyruvate dehydrogenase E1 L4: Ribosomal Protein S6 Kinase Alpha 5; Ribosomal Small component alpha Subunit, Somatic form, mitochondrial pre Subunit Pseudouridine Sy; Ribosomal Small Subunit cursor, Pyruvate dehydrogenase E1 component alpha Sub Pseudouridine Syntha; Ribosome Recycling Factor; Ribo unit, testis-specific form, mitochondrial precursor, Pyruvate some-Inactivating Protein Alpha-Trichos: Ribulose-1.5 Bis Dehydrogenase Kinase, Isozyme 2: Pyruvate dehydrogenase phosphate Carboxylase/Oxygen; Ricin; Ricin A Chain; protein X component, mitochondrial precursor, Pyruvate Right-Handed Coiled Coil Tetramer; Right-Handed Coiled Kinase; Pyruvate kinase, isozymes M1/M2; Pyruvate Kinase, Coil Trimer; RNA 3’-Terminal Phosphate Cyclase; RNA Isozymes R/L. Pyruvate Oxidase; Pyruvate Phosphate Diki Dependent RNA Polymerase: RNA Ligase 2: RNA Poly nase; Pyruvate, Orthophosphate Dikinase; Pyruvate:ferre merase Alpha Subunit; RNA-Binding Protein Regulatory doxin oxidoreductase (PFOR) in anaerobes; Pyruvate-Ferre Subunit; RNA-Dependent RNA Polymerase: RNA-Directed doxin Oxidoreductase; Pyruvoyl-Dependent Arginine RNA Polymerase: Rnase L Inhibitor; Rnase Ngr3; Rind3/ Decarboxylase; Pyst1: Quercetin 2,3-Dioxygenase; Queuine Rhoe Small GTP-Binding Protein: Rod Shape-Determining tRNA-Ribosyltransferase; Quinohemoprotein Alcohol Protein Mreb; Rop Ala2IIe2-6; Rubredoxin; Rubredoxin: Dehydrogenase; Quinolinate Phosphoribosyl Transferase: Oxygen Oxidoreductase; Ruvb; Rv3303C-Lpda; Ryanodine Quinolinic Acid Phosphoribosyltransferase: Quinone Oxi receptor 1: S-Gamma86-Beta-Mercaptoethanol-Lysozyme; doreductase; Quinone Reductase; Quinone Reductase Type S-Gamma97-Beta-Mercaptoethanol Lysozyme: S100A6; 2: Quinone-Reductase; Quinoprotein Ethanol Dehydroge S3-Rnase; Saccharopepsin; Saccharopepsin precursor, Sac nase; Rab GDP Disossociation Inhibitor Alpha; Rab6 Gitpase: charopine Reductase; S-Adenosylhomocysteine Hydrolase; RAC serine/threonine-protein kinase; Rac-Alpha Serine/ S-Adenosyl-L-Homocysteine Hydrolase; S-Adenosyl-L- Threonine Kinase; Radixin: RAF proto-oncogene serine/ Methionnine: Salicylic Acid Car; S-Adenosylmethionine threonine-protein kinase; Ramoplanin; Ran; Rap2A; Ras Decarboxylase Proen; S-Adenosylmethionine Decarboxy Related C3 Botulinum Toxin Substrate 1 I Ras-Related lase Proenzyme: S-Adenosylmethionine Synthetase; S-Ad Protein Rab-1 1A: Ras-Related Protein Rab-5A; Ras-Related enosyl-Methyltransferase Mraw; Salicylic Acid-Binding Protein Rab-7: Ras-Related Protein Rab-9A, Ras-Related Protein 2; Salivary Lipocalin; Salivary Nitrophorin; Salvia Protein Ral-A: Ras-Related Protein Sec4; Rat ADP-Ribosy officinalis; Sandostatin: Saposin B: Sari; Sarcoplasmic/Endo lation Factor-1; Rat Synapsin I: Re-Rnase6 Ribonuclease: plasmic Reticulum Calcium A: Sarcosine Oxidase; Sarcosine Reca; Reca Protein; Receptor Protein-Tyrosine Kinase Erbb Oxide: Scaffolding Dockerin Binding Protein A; Scavenger 3; Receptor-Type Adenylate Cyclase Gresag 4.1; Recombi receptor class B member 1; Scorpion Neurotoxin; Scytalone nant Lignin Peroxidase H8; Recombination Protein Recr; Dehydratase: SDHA protein: Sec14-Like Protein 2: SEC14 Red Fluorescent Protein Fp611; Redox-Sensing Transcrip like protein 3: SEC14-like protein 4: Sec18P: Secretin recep tional Repressor; Regulatory Protein Blarl; Regulatory Pro tor; Sedolisin; Seed Coat Peroxidase; Seed Lipoxygenase-3: tein Teni; Renal Dipeptidase; Renin; Renin; Replicase, Segmentation Polarity Homeobox Protein Engrai; Segrega Hydrolase Domain; Replication Protein; Resistin; Response tion Protein: Selenocysteine Lyase: Selenosubtilisin Bpn; regulator pleb: Restriction Endonuclease Bglii; Reticulon 4 Semaphorin 3A: Seminal Plasma Protein Pdc-109; Sensor Receptor; Reticulon 4 Receptor; Retinal dehydrogenase 1: Kinase Cita; Sensor Protein Fixl; Sensory Rhodopsin II; Retinal dehydrogenase 2: Retinal Dehydrogenase Type II; Sepiapterin Reductase: Serine Acetyltransferase: Serine Car Retinoic acid induced 3 protein; Retinoic acid receptor alpha; boxypeptidase; Serine Hydroxymethyltransferase: Serine Retinoic Acid Receptor Beta: Retinoic Acid Receptor hydroxymethyltransferase (mitochondrial); Serine Gamma-1: Retinoic Acid Receptor Gamma-2: Retinoic acid Hydroxymethyltransferase, Cytosolic; Serine palmitoyl receptor responder protein 1: Retinoic acid receptor RXR transferase 1: Serine palmitoyltransferase 2: Serine Protease: alpha; Retinoic acid receptor RXR-beta; Retinoic acid recep Serine racemase; Serine/Threonine Kinase 6: Serine/Threo tor RXR-gamma; Retinoid X Receptor, Beta: Retinol Bind nine Protein Kinase Tao2; Serine/Threonine Protein Phos ing Protein Complexed With Axero: Retinol Dehydratase: phatase 5: Serine/Threonine Protein Phosphatase Pp1-Gam; retinol dehydrogenase 12; retinol dehydrogenase 13; Retinol Serine/threonine-protein kinase ALS2CR7: Serine/threo binding protein I: Retinol-binding protein II: Retinol-binding nine-protein kinase receptor R2, Serine/threonine-protein protein III: Retinol-binding protein IV: Retropepsin; Reverse kinase receptor R3: Serine/Threonine-Protein Kinase Gyrase; Rifbc; Rfcs; Rhamnogalacturonan Acetylesterase; Ymr216C; Serine-Carboxyl Proteinase; Serine-pyruvate US 2015/0309021 A1 Oct. 29, 2015 56 aminotransferase: Serotonin N-Acetyltransferase; Serotrans S-transferase A2; sp|P10632ICP2C8 HUMAN Cytochrome ferrin; Serotransferrin precursor; Serratia Marcescens Ami P450 2C8 (EC 1.14.14.1); sp|P10635|CP2D6 HUMAN noglycoside-3-N-Acet; Serum Albumin; Serum albumin pre Cytochrome P450 2D6 (EC 1.14.14.1); sp|P11509|CP2A6 cursor: Serum Amyloid P Component; Serum Transferrin; HUMAN Cytochrome P450 2A6 (EC 1.14.14.1): Seryl-tRNA synthetase: Set9; Sex Comb On Midleg Cg.9495 sp|P11712|CP2C9 HUMAN Cytochrome P450 2C9 (EC Pa; Sex Hormone-Binding Globulin; Sex hormone-binding 1.14.13.80); sp|P15104|GLNA HUMAN Glutamine syn globulin precursor: Sf11-Rinase; Sfua; Sh3 Domain-Binding thetase (EC 6.3.1.2) (Glutamate-ammonia ligase) (GS); Glutamic Acid-Rich Protein: Shiga Toxin B-Chain; Shiga sp|P15289|ARSA HUMAN Arylsulfatase A precursor (EC Like Toxin IB Subunit; Shiga-Like Toxin I Subunit B; Shiga 3.1.6.8) (ASA) (Cerebroside-sulfatase) Contains: Arylsulfa Like Toxin lie B Subunit; Shikimate 5'-Dehydrogenase; tase A component B; Arylsulfatase A component C: Shikimate 5-Dehydrogenase; Shikimate 5-Dehydrogenase 2: sp|P15531|NDKA HUMAN Nucleoside diphosphatekinase Shikimate Kinase; Short Chain 3-Hydroxyacyl-Coa Dehy A (EC 2.74.6); sp|P16152|DHCA HUMAN Carbonyl drogenase; Short chain 3-hydroxyacyl-CoA dehydrogenase, reductase NADPH 1; sp|P16435INCPR HUMAN mitochondrial precursor; Short Chain Acyl-Coa Dehydroge NADPH-cytochrome P450 reductase; splP19099|C11B2 nase; Short-Chain Dehydrogenase/Reductase Family M: HUMAN Cytochrome P450 11B2; sp|P19971 ITYPH HU Shp-2: Siah-1A Protein; Sialic Acid Binding Ig-Like Lectin MAN Thymidine phosphorylase; sp|P20711|DDC HU 7; Sialidase; Sialidase 2: Sialoadhesin: Sigfl-Gfp Fusion Pro MAN Aromatic-L-amino-acid decarboxylase; tein; Sigma Factor Sigb Regulation Protein Rsbq; Signal sp|P20813|CP2B6 HUMAN Cytochrome P450 2B6 (EC Peptidase I: Signal Processing Protein; Signal Processing 1.14.14.1); sp|P21397|AOFA HUMAN Amine oxidase fla Protein; Signal Recognition Particle 9/14 Fusion Prot; Signal vin-containing; sp|P22309|UD11 HUMAN UDP-glucu Recognition Particle Protein; Signal Sequence Recognition ronosyltransferase 1-1 precursor; sp|P22310|UD14 HU Protein Ffh; Signalling Protein Spb; Silent Information Regu MAN UDP-glucuronosyltransferase: sp|P23141|EST1 lator 2; Simian Immunodeficiency Virus; Similar To Synap HUMAN Liver carboxylesterase 1 precursor (EC 3.1.1.1): totagmini/P65; Similar To Thymidylate Kinase. Sindbis sp|P27338|AOFB HUMAN Amine oxidase B; Virus Capsid Protein; Single Stranded DNA Binding Protein; sp|P27707|DCK HUMAN Deoxycytidine kinase (EC 2.7.1. Siroheme Biosynthesis Protein Met&; Siroheme Synthase: 74) (dCK); sp|P28332|ADH6 HUMAN Alcoholdehydroge Skeletal Dihydropydrine Receptor, Smad2; Small Inducible nase 6 (EC 1.1.1.1); sp|P32320ICDD HUMAN Cytidine Cytokine A20; Small inducible cytokine A23 precursor: deaminase (EC 3.5.4.5); sp|P33261|CP2CJ HUMAN Cyto Small Inducible Cytokine A5; Small Inducible Cytokine B10; chrome P450 2C19 (EC 1.14.13.80); sp|P42898 MTHR Small Nuclear Ribonucleoprotein Homolog: Small Tetra HUMAN Methylenetetrahydrofolate reductase; heme Cytochrome C; Sm-Like Archaeal Protein 1: SNAP-25; sp|P479891XDH HUMAN Xanthine dehydrogenase/oxi Sniffer Cg 10964-Pa; Sodium- and chloride-dependent creat dase; sp|P48775|T23O HUMAN Tryptophan 2,3-dioxyge ine transporter 1: Sodium- and chloride-dependent creatine nase (EC 1.13.11.11) (Tryptophan pyrrolase) (Tryptopha transporter 2: Sodium- and chloride-dependent GABA trans nase) (Tryptophan oxygenase) (Tryptamin 2,3-dioxygenase) porter 1: Sodium- and chloride-dependent neutral and basic (TRPO); sp|P50135|HNMT HUMAN Histamine N-methyl amino acid transporter B(0+); Sodium/Hydrogen Exchanger transferase to; sp|Q06278 ADO HUMAN Aldehyde oxidase 1: Sodium/potassium-transporting ATPase alpha-1 chain; (EC 1.2.3.1); sp|Q07973|CP24A HUMAN Cytochrome Sodium/potassium-transporting ATPase gamma chain; P450 24A1; splQ16696||CP2AD HUMAN Cytochrome Sodium-dependent norepinephrine re-uptake pump; Sodium P450 2A13 (EC 1.14.14.1); sp|Q6GRKOIQ6GRKO HU dependent proline transporter, Sodium-dependent serotonin MAN Cytochrome P450, subfamily IIIA, polypeptide 4: re-uptake pump; Soluble Cytochrome B562; Soluble Lytic sp|Q9NTN3S35D1 HUMANUDP-glucuronic acid; Sperm Transglycosylase Slt70; Soluble Quinoprotein Glucose Whale Metaquomyoglobin Variant H93G: Spermidine Syn Dehydrogenase; Soluble Tumor Necrosis Factor Receptor 1: thase: Spermidine/Putrescine-Binding Protein; Spermine Solute carrier family 12, member 2: Solute carrier family 12, oxidase; Sphingosine-1-phosphate lyase 1: Split-Soret Cyto member 4: Solute carrier family 12, member 5; Solute carrier chrome C; SpoOA; SpoCB-Associated GTP-Binding Pro family 12, member 6: Solute carrier family 12, member 7; tein; Spore Coat Polysaccharide Biosynthesis Prote: Spp-40: Solute carrier family 13, member 1: Solute carrier family 13, Squalene Epoxidase; Squalene-Hopene Cyclase; Squalene member 2: Solute carrier family 13, member 3: Solute carrier Hopene Cyclase; Sr Protein Kinase; S-Ribosylhomocystein family 23 member 1: Solute carrier family 23 member 2: ase; Sst 1-Selective Somatosatin: Sst 1-Selective Somatosatin Somatostatin receptor type 1: Somatostatin receptor type 2: Analog: Staphopain; Staphylococcal Enterotoxin B: Stat Pro Sorbitol Dehydrogenase; Sorting Nexin Grd 19; tein; Stem/Leaf Lectin Db58; Steroid Delta-Isomerase; Ste sp|O43272|PROD HUMAN Proline oxidase, mitochondrial roid Sulphotransferase: Sterol 14-alpha demethylase; Sterol precursor (EC 1.5.3.-) (Proline dehydrogenase); Carrier Protein 2: Sterol-4-alpha-carboxylate 3-dehydroge sp|P04746|AMYP HUMAN Pancreatic alpha-amylase pre nase, decarboxylating; Steryl-Sulfatase: Sticholysin II; cursor; sp|P04798|CP1A1 HUMANCytochrome P450 1A1 Streptavidin; Streptavidin precursor; Streptogramin A (EC 1.14.14.1); sp|P05108|C11A HUMAN Cytochrome Acetyltransferase; Stringent Starvation Protein A; Stromel P450 11A1; sp|P05177|CP12 HUMAN Cytochrome P450 ysin 3: Stromelysin-1; Structural Polyprotein; Subtilisin 1A2; sp|P05177|CP1A2 HUMAN Cytochrome P450 1A2 8321; Subtilisin 8324; Subtilisin Bpn"; Subtilisin BPN'8350; (EC 1.14.14.1); sp|P05181|CP2E1 HUMAN Cytochrome Subtilisin Carlsberg; Subtilisin Carlsberg, Type Viii; Subtili P450 2E1 (EC 1.14.14.1); sp|P05186|PPBT HUMAN Alka sin PbS2; Subtilisin-Carlsberg; Subunit C: Succinate dehy line phosphatase, tissue-nonspecific isozyme precursor; drogenase; Succinate semialdehyde dehydrogenase, mito sp|P08263|GSTA1 HUMAN Glutathione S-transferase A1; chondrial precursor. Succinylarginine Dihydrolase; sp|P08684|CP3A4 HUMAN Cytochrome P450 3A4 (EC Succinyl-CoA ligase GDP-forming beta-chain; Sucrose 1.14.13.67); sp|PO9210|GSTA2 HUMAN Glutathione Phosphorylase. Sucrose-Specific Porin: Sugar Transport Pro US 2015/0309021 A1 Oct. 29, 2015 57 tein; Sulfatase Modifying Factor 2: Sulfate Adenylyltrans 5S Subunit; Transcobalamin I; Transcobalamin II; Transcrip ferase: Sulfide Dehydrogenase; Sulfite Oxidase; Sulfite tion Antitermination Protein Nusg: Transcription Antitermi Reductase; Sulfite Reductase Hemoprotein; Sulfolipid Bio nator Lict; Transcription factor PPARgamma; Transcrip synthesis; Sulfolipid Biosynthesis Protein Sqd 1; sulfonyl tional Activator Tena; Transcriptional Regulator; urea receptor (SUR1); Sulfonylurea receptor 1: Sulfonylurea Transcriptional Regulator Nadr, Transcriptional Regulator, receptor 2: Sulfotransferase; Sulfotransferase Family, Cyto Hth 3 Family: Transcriptional Regulator, Rok Family; Tran solic, 2B, Memb; Sulfurtransferase: Superoxide Dismutase: scriptional Regulatory Protein Fixi: Transcriptional Regula Superoxide Dismutase Cu-Zn): Superoxide Dismutase Fe: tory Protein, Sir2 Fam; Transcriptional Repressor Ethr: Superoxide Dismutase Mn). Mitochondrial; Superoxide Transducin-Alpha; Transferrin Receptor Protein; Transform Dismutase Ni: Suppressor Of Tumorigenicity 14: Surface ing Growth Factor-Beta 3: Transforming Protein P21/H-Ras Layer Protein; Surface Protein; Survival Protein E: Synapsin 1: Transforming Protein Rhoa; Transhydrogenase Diii; Tran Ia: Synthetic Designed Peptide Alpha-1: Synthetic Peptide sient Receptor Potential-Related Protein; Transitional Analogue Of Shk Toxin; System Namino acid transporter 1: Endoplasmic Reticulum Atpas. Transitional Endoplasmic T Lymphocyte Activation Antigen; T3-785: T4 Lysozyme: Reticulum Atpase; Transketolase; Transketolase-like 1: Tachylectin-2, Tagatose-Bisphosphate Aldolase Agay, Tail Translation Elongation Factor Selb; Translation Initiation spike Protein; Taka Amylase; Tandem pH Domain Contain Factor Eif4E: Translation Initiation Factor If2/EifSB; ing Protein-1; Taq DNA Polymerase; Taste receptor type 1 Transpeptidase (bacterial); Transposase Inhibitor Protein member 1: Taste receptor type 1 member 2: Taste receptor From Tn5; Trans-Sialidase; Transthyretin: Transthyretin pre type 1 member 3: Tatd-Related Deoxyribonuclease; Tat-In cursor; Transthyretin Thr119Met Variant; Trehalose Operon teracting Protein Tip30. T-Cell Ecto-ADP-Ribosyltrans Repressor; Triacylglycerol Lipase, Gastric; Triacyl-Glyc ferase 2: T-Cell Ecto-ADP-Ribosyltransferase 2: T-cell sur erol-Hydrolase; Trichodiene Synthase; Trichotoxin A50E: face glycoprotein CD3 epsilon chain (CD3E); Tap-Glucose Tricorn Protease; Trifunctional enzyme alpha subunit, mito 4,6-Dehydratase; Tdp-Glucose-4,6-Dehydratase: chondrial precursor; Trigger Factor, Triggering Receptor Techylectin-5A: Tem-1 Beta Lactamase; Tem-1 Beta-Lacta Expressed On Myeloid Cel; Trihydroxynaphthalene Reduc mase; Terminal Deoxynucleotidyltransferase Short I; Teta tase; Trimethylamine Dehydrogenase; Triose Phosphate nus Toxin; Tetanus Toxin He; Tetanus Toxin Heavy Chain; Isomerase; Triosephosphate Isomerase; Triosephosphate Tetra-; Tetracycline Repressor; Tetrahydrodipicolinate Isomerase, Glycosomal; Trk System Potassium Uptake Pro N-Succinyltransferase; Tetrameric Beta-Beta-Alpha Mini tein Trka Horn; tRNA; trNACca-Adding Enzyme; trNA Protein: Tgf-Beta Receptor Type I: Tgf-Beta Receptor Type Nucleotidyltransferase; thNA-Guanine Transglycosylase: II: Th10Aox: Th1 OBox: Th1 Ox; Thaumatin I: The GTP Tropinone Reductase-I; Tropinone Reductase-II; Troponin Binding Protein Obg: The Hypothetical Protein; The Major C. Slow Skeletal and Cardiac Muscles; Troponin I, cardiac Capsid Protein Of Pbcv-1, Vp54; Thermal Hysteresis Pro muscle; Troponin T. cardiac muscle; Trp Operon Repressor; tein; Thermolysin; Thermonuclease; Thermonuclease pre Trp Repressor Binding Protein Wrba; Trp RNA-Binding cursor: Thermosome Alpha Subunit; Thiamin Phosphate Attenuation Protein Complexed: Trypanothione Oxidoreduc Synthase; Thiamin Pyrophosphokinase; Thiamine pyrophos tase; Trypanothione Reductase; Tryparedoxin II; Trypsin; phokinase; Thiamine transporter 1; Thiamine triphosphatase; Trypsin I; Trypsin II, Anionic; Trypsin Inhibitor Bgit; Trypsin Thiazide-sensitive sodium-chloride cotransporter; Thiazole Inhibitor I; Trypsin Iva; Trypsinogen; Tryptamine D recep Biosynthetic Enzyme: Thioesterase: Thioesterase I: Thiol tors (5HT2B); Tryptophan 2,3-dioxygenase; Tryptophan Peroxidase; Thiol: Disulfide Interchange Protein; Thiol-Dis 5-hydroxylase 1: Tryptophan 5-hydroxylase 2: Tryptophan ulfide Oxidoreductase Resa; Thioredoxin; Thioredoxin 5-Monooxygenase; Tryptophan Synthase Alpha Chain; Tryp reductase; Thiostrepton; Threonine Synthase: Threonine syn tophan Synthase Alpha Chain, Chloroplast; Tryptophan thase-like 1: Threonyl-tRNA Synthetase: Threonyl-tRNA tRNA Ligase: Tryptophanyl-tRNA Synthetase; Tt0826; Synthetase 1: Thrombomodulin; Thrombospondin 1: Throm Ttk003001606; Tubby Protein; Tubulin; Tubulin alpha chain; boxane A2 receptor; Thromboxane A2 synthase; Thrombox Tubulin alpha-1 chain; Tubulin alpha-2 chain; Tubulin ane synthase; Thymidine Kinase; Thymidine Kinase, Cyto alpha-3 chain; Tubulin alpha-6 chain; Tubulin alpha-8 chain; solic: Thymidine Phosphorylase; Thymidylate Kinase; Tubulin alpha-ubiquitous chain; Tubulin beta chain; Tubulin Thymidylate Synthase; Thymidylate Synthase Thyx; Thy beta-1 chain; Tubulin beta-2C chain; Tubulin beta-3 chain; roid hormone receptor alpha; Thyroid Hormone Receptor Tubulin beta-4 chain; Tubulin beta-4q chain; Tubulin-Spe Beta-1; Thyroid hormone receptor beta-2; Thyroid peroxi cific Chaperone A.; Tumor Necrosis Factor Alpha; Tumor dase; Thyrotropin receptor (Thyroid stimulating hormone necrosis factor precursor; Type 2 Malate/Lactate Dehydroge receptor); Thyroxine-binding globulin precursor; Tick-Borne nase; Type 2 Rhinovirus 3C Protease; Type 4 Pilin; Type II Encephalitis Virus Glycoprotein; Tissue-Type Plasminogen DNA Topoisomerase Vi Subunit B: Type II Quinohemopro Activator; Toho-1 Beta-Lactamase; Toll-Like Receptor 2: tein Alcohol Dehydrogena; Type III Chloramphenicol Toll-like receptor 7 precursor; Toluene-4-Monooxygenase Acetyltransferase; Type III phosphodiesterase; Type III System Protein C; Topoisomerase IV subunit A: Topoi Secretion Chaperone Syce; Type T Voltage gated calcium somerase IV Subunit B: Tora Specific Chaperone; Toxin channel; Type-1 angiotensin II Receptor (AT1); Tyrosinase Bmkk4; tr|Q17328|Q17328. CAEEL Avermectin-sensitive precursor, Tyrosine 3-Monooxygenase; Tyrosine Ami glutamate-gated chloride channel Gluc1 beta; notransferase: Tyrosine Phenol-Lyase: Tyrosine Phos tr|Q46759|Q46759 ECOLI Beta-lactamase; phatase; Tyrosine-protein kinase ABL2: Tyrosine-Protein tr|Q5VUYOIQ5VUYO HUMAN Novel protein similar to Kinase Btk; Tyrosine-Protein Kinase Itk/Tsk; Tyrosine-Pro esterases; tr|Q75NA5|Q75NA5 MUSDO GABA-gated tein Kinase Receptor Tyro3: Tyrosine-Protein Kinase Syk: chloride channel subunit; tr|Q8ITG2|Q8ITG2 CAEEL Tyrosine-Protein Kinase Zap-70; Tyrosine-protein phos GABA-A receptor subunit; Trafficking Protein Particle Com phatase, non-receptor type 1: Tyrosyl-tRNA Synthetase: plex Subunit; Trai Protein; Transaldolase; Transcarboxylase Tyrosyl-tRNA synthetase, cytoplasmic; U1A RNA Binding US 2015/0309021 A1 Oct. 29, 2015

Domain, Ubiquinol-cytochrome-c reductase complex core tein; Wunen-Nonfunctional Gfp Fusion Protein; Xaa-Pro protein I, mitochondrial precursor, Ubiquitin: Ubiquitin-Ac Aminopeptidase; Xaa-Pro Dipeptidase; Xanthan Lyase; Xan tivating Enzyme E1 1, Ubiquitin-Conjugating Enzyme E22; thine Dehydrogenase: Xanthine oxidase; Xanthine Phospho Ubiquitin-Conjugating Enzyme E2-25 Kda; Ubiquitin-Like ribosyltransferase: Xanthine-Guanine Phosphoribosyltrans Protein 7, Rub 1: Ubiquitin-Protein Ligase E3 Mdm2; Udp ferase; Xanthosine Phosphorylase; Xenobiotic 3-O3-Hydroxymyristoyl N-Acetylglucosa; Udp-Galacto Acetyltransferase: Xylanase; Xylanase 10C; Xylanase pyranose Mutase; Udp-Galactose 4-Epimerase; Udp-Galac Inhibitor Protein I; Xylanase Y; Xyloglucan Endotransglyco tose 4-Epimerase; Udp-Galactose-4-Epimerase; Udpglenac sylase; Xylose Isomerase: Xylose Reductase: Xylose Reduc Pyrophosphorylase; Udp-Glucose 4-Epimerase: Udp-Glu tase; Y177F Variant Of S. Enterica Rimla Bound To U; Yajq cose 4-Epimerase; UDP-glucose 6-dehydrogenase; Udp Protein; Ydrs33C Protein: Yeast Cytochrome C Peroxidase; Glucose Dehydrogenase; UDP-glucuronosyltransferase 1-9 Yeast Iso-1-Ferrocytochrome C; Yeco; Yfua; Yibi Protein; precursor, microsomal; Udp-N-Acetylenolpyruvoylglu Yigf Protein; Ykof: Ylmd Protein Sequence Homologue: cosamine Reductas; Udp-N-Acetylglucosamine 1-Carbox Ylr011 Wp: Ynk-Contryphan: Yvrk Protein: Zervamicin lib; yvinylt: Udp-N-Acetylglucosamine 1-Carboxyvinyl-Trans; Zinc FingerY-Chromosomal Protein; and Zinc-Alpha-2-Gly UDP-N-acetylglucosamine 1-carboxyvinyltransferase: Udp coprotein. N-Acetylglucosamine 2-Epimerase; Udp-N-Acetylglu cosamine Enolpyruvyl Tran; Udp-N-Acetylglucosamine Complete System for Analysis Enolpyruvyl Transfer, Udp-N-Acetylglucosamine Pyrophos phorylase; Udp-N-Acetylglucosamine-1-Phosphate I0083. In order to determine binding constants using Uridyltr; Udp-N-Acetylglucosamine-N-Acetylmuramyl-: X-Ray fluorescence, a system must be created that is capable Udp-N-Acetylmuramate-Alanine Ligase; Udp-N-Acetylmu of measuring the X-Ray fluorescence of the bound small ramate-L-Alanine Ligase; Udp-N-Acetylmuramoylalanyl molecule or drug. A process must exist for equilibrating the D-Glutamate- Udp-N-Acetylmuramoyl-L-Alanine: chemical with the protein, this process requires the combina D-Glutam; Udp-N-Acetylmuramoyl-L-Alanine: tion of the chemical and the protein in Solution, and incuba D-Glutamate; Udp-N-Acteylglucosamine Pyrophosphory tion of the chemical and protein solution for sufficient time to las; Ulex Europaeus Lectin II: Ultraspiracle Protein; UMP ensure that chemical equilibrium is obtained; separating the CMP kinase; Uncoordinated protein 63: Unknown Protein; unbound/excess Small molecule in a manner that does not Unknown Protein From 2D-Page: Unlocked Metal-Free Con unacceptably perturb the binding, and assaying the protein for canavalin A; Unsaturated Glucuronyl Hydrolase; Upf)230 the bound molecule, which requires that the separation be Protein Tm 1468; Upper Collar Protein: Uracil Phosphoribo obtained in less than 5 minutes at 37 degrees Celsius, and syltransferase; Uracil-DNA Glycosylase; Uracil-DNA Gly more preferably in less than one minute at 4 degrees Celsius, cosylase Inhibitor; Ure2 Protein: Urease alpha subunit; and most preferably in less than 30 seconds at 4 degrees Uricase; Uridine Diphospho-N-Acetylenolpyruvylglucosa; Celsius. In the case of XRF, one or more atoms in the chemi Uridine Phosphorylase; Uridine Phosphorylase, Putative: cal fluoresces when irradiated with X-Rays. This x-ray fluo Uridine-Cytidine Kinase 2; Uridylate Kinase; Uridylmono rescence signal can be used to determine the concentration of phosphate/Cytidylmonophosphate Kin: Urocanase Protein; the bound small molecule when proper calibration is used. Urocanate Hydratase; Urokinase; Urokinase-Type Plasmino Calibration is especially important for X-ray fluorescence, gen Activator: Uroporphyrin-III C-Methyltransferase: and is more important for X-ray fluorescence than for optical Uroporphyrinogen Decarboxylase; Vacuolar ATP Synthase fluorescence, because the X-ray penetration depths of differ Subunit C; Valyl-tRNA synthetase: Vanadium Bromoperoxi ent chemical elements vary greatly. In contrast to optical dase; Vanillyl-Alcohol Oxidase; Variant Surface Glycopro fluorescence where the optical clarity of the sample and the tein; Vascular endothelial growth factor; Vascular endothelial sample holder is easy to assess; in X-ray fluorescence, the growth factor receptor 2 precursor; Vascular endothelial X-ray attenuation in the sample and the effect of secondary growth factor receptor 3 precursor; Vascular Endothelial excitation from the Substrate and secondary excitation within Growth Factor Toxin; Vasopressin V1a receptor; Vasopressin the sample can be large and unpredictable. V1b receptor; Vasopressin V2 receptor, Vasopressin V2 I0084. The intensity of the XRF fluorescence of the small receptor, Neurophysin 2; Venom Serine Proteinase; Venom molecule can be compared in a ratio against the intensity of Serine Proteinase; Vesicular Monoamine Transporter; Vhh atoms in the protein, or the concentration of the bound mol R2 Anti-Rró Antibody; Vim-2 Metallo-Beta-Lactamase: ecule can be compared to known values from the set up of the Vimentin; Viral guanylyltransfearse; Viral RNA polymerase: binding experiments to calculate binding constants. For Virb11 Homolog:Vitamin B12 Receptor; Vitamin B12 Trans example, a protein can be incubated with an inhibitor under a port Protein Btuf, Vitamin D Binding Protein; Vitamin D range of conditions from very low binding to very high bind Receptor; Vitamin D3 Receptor; Vitamin D-Dependent Cal ing (close to 100% of the protein). The unbound ligand is cium-Binding Prote; Vitamin K Reductase; Vitamin K-de separated via gel filtration or another size exclusion tech pendent protein Z: Voltage gated sodium channel; Voltage nique, and an aliquot from each sample is spotted onto a dependent L-type calcium channel alpha-1C Subunit; substrate for XRF measurement. From the intensity of the Voltage-dependent L-type calcium channel beta-1 subunit; fluorescence, a concentration of bound Small molecule can be Voltage-dependent P/O-type calcium channel alpha-1A sub determined. A preferred method for assessing binding con unit; Voltage-dependent T-type calcium channel; Voltage stants consists of measuring the intensity of the X-ray fluo Gated Potassium Channel; Voltage-gated Sodium channel; rescence signal that is due to an atom in the bound chemical; Volvatoxin A2: Vp1 Protein; Vp39; VSV Matrix Protein: converting the measured intensity into a concentration value, V-Type Sodium ATP Synthase Subunit K: W224HVariant Of preferably using a computer, plotting the concentration as a S. Enterica Rimla Bound To U; Wbpp. Wheat Germ. Aggluti saturation isotherm; and extracting the binding constant via nin; Winged Bean Lectin; Wiskott-Aldrich Syndrome Pro fitting of this isotherm through non-linear regression. US 2015/0309021 A1 Oct. 29, 2015 59

0085. The system preferably has security systems for elec properly functional proteins are coordinated by other atoms tronic records, electronic signatures, and handwritten signa such as metals that also are detectable by XRF. There are two tures executed to electronic records to be trustworthy, reli ways these facts may be exploited: First, a known pure sample able, and generally equivalent to paper records and may be measured and the elemental composition determined handwritten signatures executed on paper for any records in to establish a baseline for metal ratios in pure samples; this electronic form that are created, modified, maintained, baseline may be compared with test samples to see if the archived, retrieved, or transmitted. The security system can purity and functionality match. Second, the correct Stoichi be conveniently provided by a biometric analysis, i.e. a ometry between coordinating metals and the proteins may be method of verifying an individual’s identity based on mea known, and the composition of the protein may be known, so surement of the individual’s physical feature(s) or repeatable a ratio between the coordinating metal and the protein may be action(s) where those features and/or actions are both unique used to provide a purity and functionality measurement. For to that individual and measurable. The security system may example, there are 24 Sulfur atoms for every two Zinc atoms in be conveniently provided by means of maintaining a closed functional insulin, there are four sulfur atoms for every two system, i.e. an environment in which system access is con Zinc atoms in functional hCGH, and there are 23 sulfur atoms trolled by persons who are responsible for the content of for every one copper atom. electronic records that are on the system. The security system may be conveniently provided by means of digital signatures, Method of Determining a Thermodynamic Binding Constant i.e. an electronic signature based upon cryptographic meth ods of originator authentication, computed by using a set of I0088 A thermodynamic binding constant between two rules and a set of parameters such that the identity of the binders may be measured by contacting a known amount of signer and the integrity of the data can be verified. It is each binder, removing unbound material, and measuring the preferable that digital signatures that do not use biometric amount of bound material using XRF. This is according the analysis should employ at least two distinct identification chemical equilibrium equation k=B1B2/B1B2, where components such as an identification code and password. It is B1 is the concentration of the first binder, B2 is the con preferable that the system be configured such that when an centration of the second binder, B1B2 is the concentration individual executes a series of signings during a single, con of the two binders bound to each other. This can be done more tinuous period of controlled system access, the first signing easily if the second binder (B2) has a unique, XRF-measur shall be executed using all electronic signature components; able atom (i.e., one that is larger than oxygen and that is not Subsequent signings shall be executed using at least one elec contained within the first binder). tronic signature component that is only executable by, and designed to be used only by, the individual; and when an EXAMPLE individual executes one or more signings not performed dur ing a single, continuous period of controlled system access, I0089. An embodiment of the present invention is each signing shall be executed using all of the electronic described in the following example. A 1 microliter sample of signature components. It is especially preferable that the sys protein or protein-chemical complex is diluted and re-sus tem be configured so that no two individuals have the same pended by adding 10 microliters of a buffer compatible with combination of identification code and password. It is also protein of interest, and Vortexed using a Thermolyne Maxi preferable that the identification code and password issuances Mix PlusTM and centrifuged using a revolutionary Science are periodically checked, recalled, or revised (e.g., to cover Microcentrifuge at between 6,000 RPM and 10,000 RPM. Such events as password aging). It is important that the system Matrix modifiers, such as acetic acid and an internal standard be configured so that following loss management procedures are added at this point. Acetic acid is used in amounts to be to electronically deauthorize lost, stolen, missing, or other equivalent to 2% of the total volume of protein sample. If an wise potentially compromised tokens, cards, and other internal standard is used, the internal standard reagent is devices that bear or generate identification code or password mixed with acetic acid to obtain an 8% solution of acetic acid information, and to issue temporary or permanent replace with internal standard. Typically, a 3:1 solution of protein to ments using Suitable, rigorous controls; and that transaction internal standard in acetic acid is mixed to end up with protein safeguards to prevent unauthorized use of passwords and/or in 2% acetic acid solution. The Volumes required for increas identification codes, and to detect and report in an immediate ing amounts of protein to create standard curve are deter and urgent manner any attempts at their unauthorized use to mined, and the amounts are mixed accordingly. One then the system security unit, and, as appropriate, to organizational aliquots desired amounts into 1.5 ml tubes (Eppendorf Safe management are used. Lock tubes rated up to 30,000 g) for each individual protein: reagent ratio. The aliquots are then Vortexed and centrifuged. A slide is prepared with the desired substrate (i.e. polypropy Method for Using XRF to Monitor Protein Quality lene, ultralene, mylar, kapton, aluminum, gold) by mounting I0086. Many proteins such as insulin, vaccines, antibodies, film on a cardboard or plastic slide mount. Using a pipette or human growth hormone (hGH), and clotting factor VIII are Syringe, protein solution is dispensed onto the Substrate. For produced by pharmaceutical companies. These proteins must a larger spot, a pipette (Eppendorf Series 2000 adjustable have high purity to be able to be used safely in humans. pipettes) is used to dispense 1 microliter of protein Solution Quality is very difficult and expensive to monitor for proteins. onto film, while the operator is careful not to fully evacuate These methods often require days to complete and significant the pipette tip. The sample is preferably deposited by stop labor. Thus, improved method of monitoring quality in pro ping at the first stop on pipette and touching the Solution bead teins is needed. on end of tip to film to release the sample without blowing air 0087. The present invention includes a method for the into sample. For a smaller spot, use a syringe (Hamilton simple, inexpensive, and rapid method of determining protein 7000.5 KHO.5ul 25/2.76"/3 REVT) and draw from 0.1 to 0.5 purity. Many proteins have atoms detectable by XRF, and microliters of sample into Syringe. Dispense sample onto film US 2015/0309021 A1 Oct. 29, 2015 60 by depressing Syringe completely and then touching film to amount of receptor typically may have units such as mol release bead onto film. The spot is dried under a heat lamp ecules, moles, moles per liter, grams, grams per liter, units of (GE Halogen SP 10° Beam 100W 120V) for 5 minutes. The activity, and the like. If the same amount of receptor is used sample is then measured using X-ray fluorescence on an for each experiment, then each net X-ray fluorescence signal EDAX Eagle III micro X-ray fluorescence system equipped may be directly compared. with a rhodium target excitation source (40 kV. 1000 uA) and a SiLi detector. Initial Screening measurements are collected 0097. In addition, the net XRF signal should be standard with an excitation beam dwell time of 100 milliseconds. ized for the concentration of chemical if receptors are Detailed analyses of individual samples are collected with an exposed to solutions having different concentrations of the excitation beam dwell time of 100 to 600 seconds. Measure chemical. ments will be standardized by normalizing the RPM intensity 0098. The standardized XRF signals obtained using this in control samples to the RPM signal in the experimental invention may then be compared in a variety of ways. One sample. standardized X-ray fluorescence signal may be subtracted 0090 Briefly, the present invention relates to using X-ray from another signal, for example. One standardized XRF fluorescence (XRF) as a probe to determine binding selectivi signal may be divided by another signal. Preferably, when ties of chemicals for receptors. The invention can also be used making comparisons among more than two signals, the sig to estimate therapeutic index of a chemical. Another aspect of nals are plotted on a graph so that comparisons among them the invention relates to providing an estimate of the relative may be easily visualized. therapeutic indices of two chemicals. 0091 An aspect of this invention is related to estimating 0099. Another aspect of this invention is related to esti the binding selectivity of a chemical to two or more receptors. mating the binding selectivity of at least two chemicals to two The receptors are preferably proteins or other biological or more receptors and comparing their selectivities. For this aspect of the invention, each chemical must include at least ligands. The receptors may be arrayed on a Substrate. Arrays one heavy element. The two or more chemicals may include that include from about 2 to about 100,000,000 receptors, the same heavy element or different heavy elements. The preferably from about 2 to about 1,000,000, and more pref receptors, preferably proteins or other biological ligands, erably from about 2 to about 100,000 are especially useful may be arrayed on a Substrate. Arrays may include from about with this invention. 2 to about 100,000,000 receptors, preferably from about 2 to 0092 Chemicals used with this invention generally about 1,000,000, and more preferably from about 2 to about include at least one element with an atomic number of nine or 100,000. A measured or calculated baseline X-ray fluores higher; these elements are referred to herein as “heavy ele cence signal would be obtained for the heavy element(s) in ments. each portion or aliquot of each receptor used with the inven 0093 Generally, a baseline XRF signal is established for a tion. After establishing the baseline X-ray fluorescence sig heavy element in each portion oraliquot of each receptor used nal, the receptors are exposed to a plurality of chemicals, with the invention. The baseline x-ray fluorescence signal preferably in solution, which are being tested for binding to may be calculated or measured. the receptors. Exposure of the receptors to the chemicals may 0094. After establishing the baseline XRF signal, the or may not result in the formation of one or more complexes receptor(s) are exposed to one or more chemicals, preferably that are referred to herein as “chemical-receptor complexes’. in Solution, that are being tested for binding to the receptor(s). Exposure of the receptor to the chemicals allows Such com Exposure of the receptor(s) to the chemical(s) mayor may not plexes to form, if they form at all, under the conditions of result in the formation of one or more complexes that are temperature, elapsed time, presence of other chemicals or referred to herein as “chemical-receptor complexes’. If an receptors, receptor concentration, chemical concentration, analog of the chemical is used, then the corresponding com and other parameters. After sufficient time has elapsed for one plex is referred to herein as an “analog-receptor complex”. If of the aforementioned complexes to form, the X-ray fluores the chemical is a drug, then the corresponding complex is cence signal due to the "heavy element (e.g. phosphorus, referred to herein as “drug-receptor complex”. Exposure of chlorine, fluorine, Sulfur, to name a few) in each chemical the receptor to the chemical/analog/drug allows such a com receptor complex is measured. After measuring the X-ray plex to form, if it forms at all, under the conditions of tem fluorescence signal due to the heavy element(s), the baseline perature, elapsed time, presence of other chemicals or recep X-ray fluorescence signal is Subtracted from this measured tors, receptor concentration, chemical concentration, and X-ray fluorescence signal; the difference is referred to herein other parameters. as the “net X-ray fluorescence signal'. The net X-ray fluores 0095. After sufficient time has elapsed for one of the afore cence signal is proportional to the amount of the chemical that mentioned complexes to form, the X-ray fluorescence signal is bound to the receptor in a chemical-receptor complex. due to the “heavy element (e.g. phosphorus, chlorine, fluo Next, each net X-ray fluorescence signal is standardized by rine, Sulfur, to name a few) in each chemical-receptor com dividing by the amount of the receptor in the portion used for plex is measured. binding to the chemical. The amount of receptor typically 0096. After measuring the XRF signal due to the heavy may have units such as molecules, moles, moles per liter, element, the baseline X-ray fluorescence signal is Subtracted grams, grams per liter, units of activity, and the like. If the from this measured X-ray fluorescence signal; the difference same amount of receptor is used for each experiment, then is referred to herein as the “net X-ray fluorescence signal'. each net X-ray fluorescence signal may be directly compared. The net X-ray fluorescence signal is proportional to the The standardized X-ray fluorescence signals may be com amount of the chemical that is bound to the receptor in a pared in a variety of ways. One standardized X-ray fluores chemical-receptor complex. Each net-X-ray fluorescence sig cence signal may be subtracted from another signal, for nal may be standardized by dividing by the amount of the example. One standardized X-ray fluorescence signal may be receptor in the portion-used for binding to the chemical. The divided by another signal. Preferably, when making compari US 2015/0309021 A1 Oct. 29, 2015

Sons among more than two signals, the signals are plotted on 0104. The invention can also be used to detect modifica a graph so that comparisons among them may be easily visu tions in proteins. alized. 0105 Examples of such detectable protein modification 0100. The invention also relates to drug manufacturing. include, but are not limited to, post-translational modification The drug manufacturing process includes the determination of proteins, phosphorylation of proteins, and dephosphoryla of which chemical or chemicals bind to a type of receptor tion of proteins. Detecting these modifications would typi referred to in the art as a "druggable target, how strongly they cally begin by first establishing a baseline XRF signal for a bind, whether or not they bind selectively (i.e. whether or not heavy element in each portion or aliquot of each protein, and they also bind undesirably to other receptors), and for those then exposing the protein to conditions that may result in that bind selectively to what degree the selectivity is. After protein modification. Afterward, the XRF signal due to the identifying a drug that binds strongly and selectively to a heavy element would be measured. The baseline X-ray fluo desired druggable target, the drug is synthesized in large rescence signal is then subtracted from this measured XRF quantities. signal, and the difference is the “net XRF signal'. The net XRF signal would be standardized by dividing by the amount 0101 The invention can be used to estimate the binding of the receptor in the portion being tested. The standardized selectivity of a chemical to one or more receptors under XRF signals may then be compared. different conditions. For example, the binding affinity of a 0106. This invention may be used to estimate the thera chemical in one solution can be compared to its binding peutic index of a chemical/drug/analog. The therapeutic affinity in a different solution. The chemical would include index is a measure of drug selectivity that is ordinarily calcu one or more heavy elements. The effect that other compo lated from data obtained from experiments with animals. The nents that do not have a heavy element in either solution have therapeutic index of the drug is typically defined as the ratio on the binding affinity of the chemical could be inferred by of two numbers, LD50/ED50, where LD50 is the dose of a observing any observed differences in binding affinity of the drug that is found to be lethal (i.e. toxic) for 50 percent of the chemical. population of animals used for testing the drug, and ED50 is 0102 Some aspects of the invention are related to person the dose of the drug that is found to have a therapeutic effect alized medicine, and involve estimating the relative effective for 50 percent of that population. More broadly, it is a mea ness of two or more drugs. This may involve, for example, sure of the how much of the drug is needed to produce a estimating the binding selectivities of two (or more) drugs to harmful effect relative to the amount needed to produce a two (or more) receptors. The receptors may be proteins or beneficial effect. The ratio LD50/ED50 is therefore, a mea other biological ligands, typically derived from a medical sure of the approximate “safety factor” for a drug; a drug with patient or a participant in a clinical drug trial. For medical a high therapeutic index can presumably be administered patients, there may be a question about which drug to use to with greater safety than one with a low index. The invention treat the patient, and the invention can be used to guide the may be used to provide an estimate of the therapeutic index doctor toward prescribing a specific drug for treatment. For by, for example, measuring the binding affinity of a chemical applications related to personalized medicine, these types of to a receptor associated with side effects and to a receptor drugs would include at least one heavy element. For drugs associated with efficacy. The ratio of the measured binding that are being compared, the drugs might include the same affinities (efficacy divided by side effects) provides an esti heavy element, or different heavy elements. Each drug may mate of the “therapeutic index. For an example of using be a single chemical, or may be a mixture of chemicals. If the DNA arrays with optical fluorescence high-throughput drug is a mixture of chemicals, then one of the active ingre screening to estimate a therapeutic index, see R. Ulrich and S. dients of the drug must have a heavy element. H. Friend, “Toxicogenomics And Drug Discovery: Will New 0103) An aspect of this invention is related to estimating Technologies Help Us Produce Better Drugs?.” Nature the binding affinity of a chemical to one or more receptors, Reviews Drug Discovery, vol. 1, pp. 84-88 (2002), incorpo preferably proteins or other biological ligands that are rated herein by reference. arrayed on a substrate. The chemical would include at least 0107 An example relating to the use of the invention to one heavy element. First, a baseline XRF signal for the heavy obtain a simple estimate of a therapeutic index may involve element in each portion or aliquot of each receptor would be exposing an array of the proteins 5HT2A and NAa1 to a obtained, either by measuring or calculating the baseline sig solution of olanzapine. Assuming that the 5HT2A and NAa1 nal. Afterward, the receptor(s) would be exposed to one or of the array are present in equal amounts, the net sulfur XRF more chemicals. Chemicals that bind to the receptors result in signal due to the olanzapine should be between 1.6 and 20 the formation of chemical-receptor complexes that are detect times higher for olanzapine bound to 5HT2A than for olan able by measuring the XRF signal due to the heavy element in Zapine bound to NAa1. each chemical-receptor complex. This measured X-ray fluo 0108. Another example relating to the use of the invention rescence signal is obtained by using X-ray excitation photons to obtain an estimate of a therapeutic index is now described. with an energy of at least 300 eV, which generate emission A therapeutic index for a non-steroidal anti-inflammatory photons having a fluorescence lifetime that is less than the drug (commonly referred to as an “NSAID) may be obtained dead time of the X-ray detector. After measuring the X-ray by exposing an array that includes the proteins COX-1 and fluorescence signal due to the heavy element, the baseline COX-2 to a solution of meloxicam. Assuming the COX-1 and XRF signal is subtracted from this measured x-ray fluores COX-2 are present in equal amounts, the net XRF signal due cence signal, yielding the net XRF signal, which is propor to sulfur in the meloxicam should be about 10 times higher for tional to the amount of the chemical that is bound to the meloxicam bound to COX-2 than for meloxicam bound to receptor in a chemical-receptor complex. The net XRF signal COX-1. would then be standardized. The standardized XRF signals 0109 Yet another example relating to the use of the inven may then be compared. tion to obtain an estimate of a therapeutic index may involve US 2015/0309021 A1 Oct. 29, 2015 62 exposing an array that includes COX-1, COX-2, and proteins proteins, for example COX-1 and COX-2 derived from a derived from hepatocytes, to a solution of meloxicam. blood sample from a patient, may be exposed to the drug Assuming the COX-1 and COX-2 are present in equal meloxicam. The binding affinities of meloxicam to COX-1 amounts, the net XRF signal due to the sulfur in the meloxi and COX-2 are known. Therefore, the net XRF signal due to cam should be about 10 times higher for meloxicam bound to sulfur in the meloxicam that is bound to COX-1 versus that for COX-2 than for meloxicam bound to COX-1. Any XRF signal COX-2 may indicate the relative amounts of COX-1 and due to sulfur in meloxicam bound to hepatocyte-derived pro COX-2 in the blood sample. teins may suggest that the meloxicam may concentrate in the 0113. The invention may also be used to develop optically liver and may be cause for further analysis of the toxicity of fluorescent immunoassay diagnostics by comparing the meloxicam. selectivity of a non-fluorescently tagged chemical to a fluo rescently-tagged chemical. XRF would be used to determine the binding selectivity of the non-tagged chemical, and either CH3 XRF or optical fluorescence would be used to determine the OH O S \ binding selectivity of the tagged chemical. If the selectivities of the tagged and untagged chemicals are proven to be simi lar, then the tagged-chemical may be used as the diagnostic N H-S N with the expectation that its binding properties are acceptably N similar to those of its untagged counterpart. 7, NCH, 0114 Binding selectivity information provided by the / O present invention is also important for uses related to Mate rials Science. Binding selectivity information can be used, for Meloxicam example, in designing high performance water filters, such as water filters that remove contaminants such as lead, arsenic, 0110 Binding selectivity information provided by the arsenate anions, or pesticides, from contaminated water. invention is important for understanding the properties of Water filters can be manufactured with receptors that have a drugs. It is also important for other applications such as for high affinity for a particular contaminant (lead, for example) medical diagnostics. For medical diagnostics, the ratio of the and a low affinity for other contaminants (iron, for example) binding affinity of probe molecules (i.e. molecules that selec in the contaminated water. By passing the contaminated water tively bind to proteins associated with a particular disease) for through this filter, the lead is removed while the iron isn’t. The a disease marker (i.e. a protein in a biological sample, Such as beneficial effect of not removing the iron is that the iron, a blood or sputum sample, from a patient having that disease, which is present in much larger amounts than the lead is, does which indicates that a disease is present) vs. the binding not overload the filter as the contaminated water is being affinity of probe molecules for other proteins not associated treated. Overloading would mean that the filter would have to with that disease, is related to the accuracy of a medical test. be replaced often. If the probe molecules bind to the disease marker, then the 0115 The binding selectivity, i.e. the ratio of binding medical diagnostic test provides a true positive result. How affinity of a chemical to one receptor versus the binding ever, when the probe molecules bind to other proteins besides affinity to a different receptor, may be performed as follows. the disease marker, then the medical diagnostic test provides First, a baseline X-ray fluorescence is obtained. This baseline a false positive result. The above ratio provides a measure of may be obtained by, for example, measuring the X-ray fluo the true positive relative to the false positive, which is related rescence (XRF) signal due to a particular element from each to the accuracy of the medical test. receptor in the absence of any added chemical. This is an 0111 Diagnostic tests that use standard immunoassay empirical measurement that establishes the baseline XRF methods require fluorescently-tagged chemicals. It has been signal for that element for each receptor. Alternatively, the previously pointed out that it is generally assumed that the baseline XRF can be calculated for each receptor. This cal attachment of a fluorescent tag to a chemical only serves to culation requires knowledge of the amount of that element make visible the otherwise invisible chemical, and does not that is present in the receptor and a calibration factor to alter its binding properties. However, Small changes to the convert an amount of that element into an XRF intensity structure of a chemical could affect its function, so this signal; the amount of the element believed to be present may assumption that tagged chemicals have the same binding then be multiplied by the calibration factor to provide a cal affinity and selectivity as their untagged counterparts may not culated XRF signal. The calibration factor may be obtained be a valid one. The tagged chemicals are structurally different by measuring a set of standards containing that element, and from their untagged counterparts, and these structural differ calculating the signal as a function of the amount of the ences could affect their binding affinities and selectivities. element. Preferably, the XRF baseline is a measured baseline. Since binding affinities and selectivities derived using tagged 0116 For cases where there is a low probability that there chemicals are suspect, the binding properties of receptors and is an element in common between the receptor and the chemi chemicals should be evaluated using the untagged chemical cal and where that element is used for the XRF measurement, or receptor and not with a tagged chemicals. This is not a foreknowledge of the composition of the receptor(s) may be possible with standard immunoassay methods, which require sufficient. For obtaining the XRF baseline for a set of organic fluorescently-tagged chemicals. By contrast, the present receptors that are being screened for their binding affinities invention allows an existing, untagged chemical (or analog or for a metal (e.g. lead) or for a chemical that has XRF-mea drug) to be used as the probe molecule. Surable chemical elements (e.g. phosphorus, chlorine) that 0112 An example of using the invention for a diagnostic are known not to be present in the set of organic receptors, for test for inflammation, for example, may include exposing example, the baseline XRF signal of the receptors may be spatially separated proteins to a drug. Spatially separated calculated or assumed to be negligible. For a Sulfur-contain US 2015/0309021 A1 Oct. 29, 2015 ing receptor (e.g. avid in) and a phosphorus-containing Snyder, “Protein Chip Technology (Review). Current Opin chemical (e.g. biotin-DHPE, or N-(Biotinoyl)-1,2-dihexade ion in Chemical Biology, Vol. 7, pp.55-63, (2003); G. Mac canoyl-sn-glycero-3-phosphoethanolamine triethylammo Beath and S. L. Schreiber, “Printing proteins as microarrays nium salt), for example, the XRF signal of phosphorus from for high-throughput function determination. Science, Vol. the avidin may be assumed to be negligible, and any phos 289, pp. 1760-1763, (2000); E. T. Fung et al., “Protein bio phorus XRF signal measured may be assumed to be due to the chips for differential profiling. Current Opinion in Biotech biotin-DHPE. nology, Vol. 12, pp. 65-69, (2001); T. Kukar et al., “Protein 0117. A receptor is preferably localized and separated Microarrays to Detect Protein-Protein Interactions Using Red from other receptors. Localization concentrates the receptor and Green Fluorescent Proteins.” Analytical Biochemistry, and any chemical bound to the receptor, therefore improving vol.306, pp. 50-54, (2002); G.Y. J. Chen et al., “Developing the sensitivity of the XRF technique. This sensitivity a Strategy for Activity-Based Detection of Enzymes in a improvement due to localization for XRF contrasts with other Protein Microarray.” ChemBioChem, pp. 336-339, (2003); K. techniques, such as optical fluorescence. Optical fluorescence Martin et al., “Quantitative analysis of protein phosphoryla may be affected by fluorescence quenching, fluorescent reso tion status and protein kinase activity on microarrays using a nance energy transfer, photobleaching, and the like. Fluores novel fluorescent phosphorylation sensor dye.” Proteomics, cence quenching diminishes the optical signal and therefore Vol. 3, pp. 1244-1255, (2003); J. Burbaum and G. M. Tobal, decreases sensitivity. Fluorescence resonance energy transfer “Proteomics in drug discovery. Current Opinion in Chemi alters the wavelength of the fluorescence signal. Photobleach cal Biology, Vol. 6, pp. 427-433. (2002); I. A. Hemmilä and P. ing degrades the fluorescence signal over time and exposure Hurskainen, “Novel detection strategies for drug discovery.” to light. By contrast, XRF is not subject to fluorescence Drug Discovery Today, Vol. 7, pp. S150-S156, (2002). The quenching because the energy differences between the arrays described in these papers may be used with the present ground state and the excited states are much greater for XRF invention. than for optical fluorescence. Energy loss via optical fluores I0122. In order to allow mass transport of a chemical to all cence may occur through non-radiative pathways that do not parts of a receptor deposited on an area of the Substrate, it is produce an optical signal. XRF Sometimes is subject to fluo preferable that there be some inhomogeneity of the receptor rescence resonance energy transfer (see, for example, J. Sher in that deposited area. Regarding the inhomogeneity of the man, Spectrochimica Acta, vol. 7, pp. 283-306 (1955)), incor deposition, the amount of receptor on a 1 square micron porated by reference herein). Optical fluorescence usually portion of the deposited area should be different from the requires an unsaturated functional group (a carbonyl group, a amount of receptor on another 1 square micron portion by at dienyl group, an aromatic group, etc.) with an electron that least 10 percent. An alternative way of comparing the amount can be excited from the ground electronic state to an excited of inhomogeneity of deposited receptor on an area involves state with non-ionizing radiation (ultraviolet, visible, and measuring the amount of chemical in each 1 Square micron near-infrared radiation). This excitation forms a reactive elec pixel within the deposited area, finding the average amount of tron-hole pair that can chemically react with neighboring the chemical per square micron over the entire deposited area, molecules, or within the same molecule. This chemical reac and calculating the standard deviation of the amount of the tion degrades the optically fluorescent chemical functional chemical per square micron pixel. The standard deviation of groups, which diminishes the optical fluorescence during the amount of the chemical per pixel divided by the average degradation. In contrast, XRF employs ionizing radiation for amount of the chemical per square micron of area should be exciting inner core electrons of atoms. The chemical environ greater than about 0.1% and preferably greater than about ment of the excited atoms does not have a significant effect on 19/6. the XRF signal. Atoms are not degraded like chemical func I0123 Receptors may be inhomogeneously deposited by tional groups, so photobleaching does not occur in XRF. attaching the receptors to polystyrene beads, for example. 0118 When the chemical binds to a localized receptor, the The points of attachment on polystyrene beads are random chemical also becomes localized. In order to maximize the and non-uniform. The attachment of receptors to beads is XRF efficiency, it is preferable that the dimensions of the described in U. S. Patent Application 20030027129 entitled receptor match those of the spot generated by the excitation “Method for Detecting Binding Events. Using Micro-X-ray beam. Fluorescence Spectrometry,” which was published on Feb. 6, 0119 For expensive receptors, it is preferable that the 2003, which describes the use of beads that were about sample of receptor be as small as possible. It is therefore 80-120 micrometers in diameter, with an average diameter of preferable that the receptor sample belocalized within an area about 100 micrometers. about 1 mm or less, and within a volume of 1 mm or less. 0.124. The measured binding affinity of a chemical to a 0120 When multiple receptors are being tested, they are receptor depends on the chemical environment and tempera each deposited in separated portions on a Substrate. The por ture. More specifically, the binding affinity of a chemical to a tions are spatially separated from each other so that the XRF receptor does not change when the chemical environment excitation beam excites only one portion, and any chemical does not change. External factors, however, may have an bound to the portion of receptor, at a time. effect on the measured binding affinity. 0121 Receptor arrays such as analytical protein arrays, 0.125. The free energy of binding of a chemical to a recep functional protein arrays, analytical DNA arrays, functional tor may be represented by the following equation: DNA arrays, analytical RNA arrays, functional RNA arrays, arrays derived from clinical patients, arrays derived from AG=AH-TAS participants in clinical trials, tissue samples, tissue arrays, where AG is the change in the Gibbs free energy of the cellular arrays, and cellular samples may be used with the binding reaction, AH is the change in enthalpy of the binding present invention. These and other types of arrays have been reaction, T is the temperature, and AS is the change in entropy described in detail in the following papers: H. Zhu and M. of the binding reaction (see, for example: Gordon M. Barrow, US 2015/0309021 A1 Oct. 29, 2015 64

Physical Chemistry, 5th Ed., McGraw-Hill, NY, 1988, Chap chemical B and chemical C are exposed to receptor 1 and ter 7). The temperature should be constant to avoid changing receptor 2, and the baseline is subtracted from the XRF sig the TAS term, because changes to the TAS term would intro nals after exposure, the similarity of the binding selectivities duce temperature derived artifacts into the measurements. AG can be assessed. For this example, chemical A and chemical B will also be affected by, for example, the presence of materials are both are selective for Receptor 1 versus Receptor 2 by that coordinate to the receptor and/or chemical. Bonds that about an order of magnitude, while chemical C is not selective form between these materials and the receptor and/or chemi for Receptor 1 versus Receptor 2, as shown in TABLE 2 cal must be broken before the chemical can bind to the recep below. tor. Energy is required to break these bonds, and this energy will be factored into AG of the equation, and will make AG of TABLE 2 binding between the chemical and receptor appear Smaller than it actually is. Therefore, in order to obtain binding affini Receptor 1 Receptor 2 ties for a chemical with multiple receptors, the chemical Chemical A 10 1 environment for the receptors and chemicals should not Chemical B 11 1 change. In addition, the temperature should not vary between Chemical C 1 1 measurements and all measurements should be taken at Sub stantially the same temperature, i.e. within +3°C., and more 0.130. This aspect of the invention may be exemplified preferably within +1° C. with specific receptors having known properties. For 0126 For drug development, the chemical environment example, Receptor 1 may be exemplified by COX-1 (inhibi preferably approximates that of the physiological environ tion of which is associated with intestinal ulcers) and Recep ment. Useful drugs should perform optimally at, or near, tor 2 may be exemplified by COX-2 (inhibition of which is physiological temperatures. One aspect of this invention is associated with anti-inflammation). related to determining binding selectivities at or near the I0131 TABLE 3 below includes selectivity data for sample temperature of the body, which is about 37 degrees Celsius. canine COX-1 and COX-2 (see, for example: Christopher Preferably, the exposure of receptors to chemicals/analogs/ Jones, Practical COX-1 and COX-2 Pharmacology: What's it drugs according to this invention is typically performed at a All About?” which can be found at the website http://www. temperature of from about 15 degrees Celsius and about 45 vetmedpub.com/cp/pdf/symposium/nov 1.pdf). degrees Celsius, and more preferably is between 32 degrees Celsius and 42 degrees Celsius, and most preferably at a TABLE 3 temperature of about 37 degrees Celsius. 0127. Useful drugs should also perform optimally in the COX-1 COX-2 presence of other, potentially interfering chemicals that are Celecoxib 1 9 also present in a physiological environment. The therapeutic Meloxicam 1 10 index provides an estimate of the performance of a drug. Ketoprofen 1 .6 Simple estimates of a therapeutic index may be obtained for New Drug (goal) 1 >10 a chemical in a solution that does not contain potentially interfering chemicals. More realistic estimates of the thera (0132 TABLE3 compares the selectivity of CELECOXIB, peutic index are obtained when the chemical is in an environ MELOXICAM, and KETOPROFEN for the receptors ment that more closely approximates biological conditions. COX-1 and COX-2. According to TABLE 3, CELECOXIB An example of such an environment is a buffered solution that has a selectivity of 9 to 1 in favor of COX-2 versus COX-1, contains chemicals that are typically found in biological and MELOXICAM has a selectivity of 10 to 1 also infavor of organisms (e.g. blood-borne ligands). COX-2 versus COX-1, while KETOPROFEN has a selectiv 0128. For medical purposes, such as the development of ity in favor of COX-1 versus COX-2 of only 1 to 0.6. Drug new therapeutic chemicals (i.e. drugs), the ratio of the binding development involves finding/developing new drugs that affinity of a chemical to the target receptor versus the binding have Superior selectivities (i.e. Superior therapeutic indices) affinity of that chemical to one or more other receptors (which Versus existing drugs. The discovery/development of more is an estimate of the therapeutic index according to the present selective drugs than those shown in TABLE 3 may involve invention) is important to know. It is also important to know measuring the selectivity of a new drug and comparing its the binding affinity and the therapeutic index estimate of the selectivity to that of the more effective drugs e.g. CELE chemical for these receptors in typical chemical environ COXIB. The goal would be to develop a new drug with ments and in the presence of common biological ligands. The superior selectivity for COX-2 over COX-1 as compared to binding affinity may be measured in the presence of for CELECOXIB. example, blood-borne proteins, ligands, fats, triglycerides, I0133) A new drug should be at least one percent more and fatty acids. These proteins and ligands may function as a selective, and more preferably more than 5 percent more proxy for distribution of the chemical between the circulatory selective than an existing, effective drug. system and cells. Fats, triglycerides, and fatty acids may I0134. An example that demonstrates how the present function as a proxy for the distribution of the chemical invention may be used to provide information related to the between fat cells (or cell membranes) and the target receptor, selectivity of a plurality of receptors to a chemical is now as well as for passive transcellular transport. described. The chemical is the lead ion. For this example, a 0129. Another aspect of the present invention is related to library consisting of 400 tripeptide receptors was prepared by determining whether or not a first chemical and a second split-pool synthesis. Each receptor of the library was chemical have similar selectivity for binding to each of a covalently attached to a spherical polystyrene resin particle. plurality of receptors. For example, if a baseline is obtained The particles had an average size of about 100 microns. The for receptor 1 and receptor 2, and then if chemical A and library of receptors was exposed to an aqueous solution, US 2015/0309021 A1 Oct. 29, 2015

having a pH of 7, of lead nitrate (7 mM). A portion of the determine which of the two is the more selective. This may library of receptors was then analyzed by XRF. The XRF involve mixing the first chemical with one or more receptors, intensity and relative binding affinity of three of the receptors allowing a receptor-chemical complex to form, and then mea for lead ion are shown in TABLE 4 below. Suring the amount of receptor-chemical complex formed using XRF, then repeating these steps with the analog. Any TABLE 4 complexes that form with the analog would also be measured by XRF. This may be repeated with additional analogs of the Receptor Pb XRF Signal Relative Binding Affinity first chemical. Receptor 4.1 991.93 111919349 0.138. There are several criteria that may be used to deter Receptor 4.2 924.94 1.04360875 mine whether chemicals are analogs of the first chemical. One Receptor 4.3 886.29 1 criterion is whether the first chemical and another chemical have one or more properties in common. The similarity may 0135 Another example that demonstrates how the present be determined by computational modeling, such as a "Quan invention may be used provide information related to the titative Structure Activity Relationship' (QSAR, see, for selectivity of a plurality of receptors to two chemicals is now example: R. Perkins et al., “Quantitative Structure-Activity described. The two chemicals are the leadion and the ironion. Relationship Methods: Perspectives on Drug Discovery and For this example, a library consisting of 400 tripeptide recep Toxicology. Environmental Toxicology And Chemistry, Vol. tors was prepared by split-pool synthesis. Each receptor of the 22, pp. 1666-1679 (2003); and T. W. Schultz et al., “Quanti library was covalently attached to a spherical polystyrene tative Structure-Activity Relationships (QSARs) in Toxicol resin particle. The particles had an average size of about 100 ogy: a Historical Perspective” THEOCHEM, vol. 622, pp. microns. The library of receptors was exposed to an aqueous 1-22 (2003), both incorporated by reference herein). solution (pH 7) of lead nitrate (7 mM) and iron nitrate (7 mM). 0.139 QSAR models compare the charge, polarity, hydro A portion of the receptors was then analyzed by XRF. The gen-bonding donor/acceptor pattern, lipophilicity, Volume, XRF intensity and relative binding affinity of two of the and other properties of chemicals to determine whether two or receptors for lead ion and iron ion are shown in TABLE 5 more chemicals are likely to have similar binding properties below. for a receptor. 0140. Another criterion for determining whether two TABLE 5 chemicals are analogs of each other relates to whether the two chemicals have a common functional group and where that Pb XRF Intensity/ functional group is attached. The following clarifies what is Pb XRF Intensity Fe XRF Intensity Fe XRF Intensity meant by the term analog. Benzene, methylbenzene, ethyl Receptor 5.1 2295 505 4.54 benzene, isopropylbenzene, and n-propylbenzene, for Receptor 5.2 13092 252 51.95 example, are analogs of each other. Olanzapine and trifluo romethylolanzapine are analogs of each other. Methyl salicy 0136. The first entry of TABLE 5 relates to data obtained late, ethyl salycylate, phenyl salicylate, N-methyl salicyla for the receptor labeled receptor 5.1, and the second entry mide, N,N-dimethylsalicylamide, and salicylic acid are relates to XRF data obtained for the receptor labeled receptor analogs of each other. The aromatic compounds 4-chlorotolu 5.2. The Pb XRF intensity of receptor 5.1 is 2295, the FeXRF ene, and 3-chlorotoluene are analogs of each other. Cyclo intensity, is 505, and the ratio of Pb XRF intensity to the Fe pentane, cyclohexane, and cycloheptane are analogs of each XRF intensity, which indicates the selectivity of receptor 5.1 other. Benzene, pyridine, and thiophene are analogs. Ethyl for binding to lead versus binding to iron, is 4.54. Similarly, benzene and methoxybenzene are analogs of each other. the Pb XRF intensity for receptor 5.2 is 13092, the Fe XRF Ethyl benzene, ethenyl benzene (i.e. styrene), and ethynyl intensity is 252, and the ratio of the Pb XRF intensity to the Fe benzene (i.e. phenyl acetylene) are analogs of each other. XRF intensity is 51.95. The ratio of 51.95 to 4.54, which More generally, enantiomers of a chemical are analogs of equals 11.4, indicates that receptor 5.2 is 11.4 times as selec each other, diastereomers are also analogs of each other. tive for lead than receptor 5.1 is. 0.141. A third criterion for determining whether two 0.137 For drug development, it is desirable to understand chemicals are analogs is related to whether the two chemicals how the drug will bind to a receptor. Drug development may share atoms having the same connectivity, where the term involve systematically varying the chemicals being tested for connectivity is meant to describe the shortest path of atoms their affinities to one or more receptors, in order to optimize that connect two functional groups. For example, 1,2-diclo the binding affinity of one of these chemical analogs to a robenzene and 1,2-dinitrobenzene have the same connectiv desired receptor and to optimize the therapeutic index, i.e. the ity between the two functional groups; the connectivity ratio of the affinity of the chemical to a desired receptor to the between the chlorine groups in the chemical 1,3-dicloroben affinities of that chemical to undesired receptors. This sys Zene is the same as the connectivity between the two nitro tematic variation of chemicals for drug development is some groups in the chemical 1.3-dinitrobenzene. times referred to as a drug development analog program. The 0.142 Preferably, a chemical being tested for binding with development of a potential new drug using analogs and XRF a receptor according to the present invention includes at least according to the present invention involves determining the one chemical element having an atomic number of nine or binding affinity of a potential drug to multiple receptors, and higher. Excitation photons used with the present invention comparing these binding affinities to provide selectivity should have energies of at least 300 eV. The excited atoms information. This process is completed for a first chemical, should have a fluorescence half-life of 10 ns or less. The and then completed for a second chemical that is an analog of importance of having Such a short fluorescence lifetime is the first chemical. Then the binding affinities and selectivities related to the “dead time', which is a period of time after the for the first chemical and for its analog are compared to X-ray fluorescence is measured. No additional X-ray fluores US 2015/0309021 A1 Oct. 29, 2015 66 cence can be measured during the dead time. Preferably, the fluorescence microscopy. The difference in sulfur content fluorescence lifetime of the type of atom being measured between the two measurements is related to the binding of should be less than the dead time of the detector. More pref olanzapine to receptors the array. erably, the fluorescence being measured should have a fluo 0.147. It is believed that the therapeutic effects of olanza rescence half-life of 10 nanoseconds (ins) or less, and most pine are a result of olanzapine binding to several receptors, preferably the fluorescence being measured should have a including the human serotonin 5HT2A receptor (see, for half-life of 100 picoseconds (ps) or less. example: http://www.gpcr.org). Toxic effects are believed to 0143. If a mixture of chemicals, as opposed to a single occur as a result of olanzapine binding to alpha-1-adrenergic chemical, is used, only one of the chemicals is required to (NAa1) receptors (see, for example: “Olanzapine have a heavy element. If more than one has a heavy element, (ZyprexaR) in Clinical Toxicology Review, Vol. 18, no. 2, and the heavy element is the same element, then a baseline March 1997. The pKi (where pKi=-log Ki, where Ki is the XRF measurement of the binding affinity of each of the inhibition equilibrium constant) of olanzapine for 5HT2A is chemicals with that element to one or more receptors should reported to be 7.8-8.9, while the pKi of olanzapine for NAa1 be obtained. The measurements may be obtained sequen is reported to be 6.3-7.6. If the binding of the chemical to a tially, e.g. by measuring the XRF spectrum of each receptor of receptor also inhibits that receptor, then the pKa and the pKi an array, then exposing the receptors of the array to a chemi of the chemical have the same value. cal, then measuring the XRF spectrum of the array after exposure to the chemical, then adding another chemical, then 0.148. The selectivity of binding of olanzapine to NAa1 measuring the XRF spectrum of the array after exposure to versus 5HT2A may be measured by first preparing an array of the other chemical. This process may be repeated for as many NAa1 and 5HT2A on a substrate, then measuring the baseline chemicals as desired. Sulfur content of each enzyme using XRF, then contacting the 0144. After measuring binding affinities between recep arrayed enzymes with a solution of olanzapine, and then tors and chemicals, the receptors may be analyzed in terms of measuring the total amount of Sulfur using XRF, and then any of their particular structural of functional features than subtracting the baseline amount of sulfur from the total may promote binding or inhibit binding. A variety of analyti amount of Sulfur, which indicates the amount of olanzapine cal techniques useful for performing this type of analysis that binds to each enzyme. include spectroscopy (e.g. infrared spectroscopy, fluores 0149 From the binding data obtained for olanzapine, it cence spectroscopy), spectrometry (e.g. mass spectrometry, appears that chemicals useful as drugs form a complex with at nuclear magnetic resonance spectrometry, surface plasmon least one receptor that has a pKi of greater than 5. resonance), functional assays, and the like. 0150. According to the invention, the X-ray excitation 0145 The binding properties of a receptor to a chemical beam is used to excite one receptor at a time. The temperature may be correlated with a positive, or with an adverse, of the array should be kept constant so that each X-ray mea response of an animal, clinical trial participant, or medical Surement is performed at the same temperature, or at nearly patient. For the case of clinical trial participants, for example, the same temperature (+3 degrees Celsius). For the case of those participants who respond adversely to the drug candi measuring the binding of olanzapine to each receptor, for date may have a different receptor affinity or selectivity than example, each measurement should be performed at the same participants that respond positively to the drug candidate. temperature or at nearly the same temperature because the 0146. One aspect of the invention may be demonstrated binding affinity changes as the temperature changes. using an array prepared as described in Kukar et al., “Protein 0151. After the selectivity of the olanzapine for binding to Microarrays to Detect Protein-Protein Interactions Using Red 5HT2A and to NAa1 is measured, additional chemicals can and Green Fluorescent Proteins.” Analytical Biochemistry, be similarly measured in order to determine whether any vol.306, pp. 50-54 (2002). The baseline sulfur content of the other chemical has a better selectivity for 5HT2A versus receptors of the array would be measured using X-ray fluo NAa1. A chemical such as the trifluoromethylolanzapine, rescence spectroscopy. Afterward, the receptors of the array which has the structural formula would be exposed to a solution of olanzapine, which has the structural formula / /CH 3 r s N NS - NS -( f S CF -( ) H f S CH3 trifluoromethylolanzapine H Olanzapine for example, may be screened against the same receptor array, or screened against another receptor array having the same and the chemical formula C7HoNS (olanzapine is the proteins, and the binding selectivity for trifluoromethylolan active ingredient in ZYPREXAR). Afterward, the sulfur con Zapine can be measured. If the selectivity of trifluoroolanza tent of each receptor would be measured again using X-ray pine for 5HT2A versus NAa1 is greater than the selectivity of US 2015/0309021 A1 Oct. 29, 2015 67 olanzapine is, then trifluoromethylolanzapine would be measurements for each protein spot is then quantified. This expected to be a Superior drug. difference is related to the binding of olanzapine to proteins in 0152 Another aspect of the present invention relates to the the array. determination of the kinetic parameters of an interaction 0158. In order to measure the rate of binding of olanzapine between a chemical and a receptor. to 5HT2A, a solution of olanzapine may be contacted with a 0153. After the baseline is measured or calculated for a 5HT2A. The 5HT2A may be in solution, or as part of an array receptor, as previously described, the receptor is exposed to a of proteins, or associated with a substrate. If the 5HT2A is in chemical at a particular temperature. The temperature is pref Solution, it should be immobilized in Some manner, such as in erably a fixed temperature, preferably between about 27 the manner described in U.S. patent application Ser. No. degrees Celsius and about 47 degrees Celsius and more pref 10/621,825 entitled “Method and Apparatus for Detecting erably is a fixed temperature between about 32 degrees Cel Chemical Binding”. Any increase in the sulfur XRF signal, sius and about 42 degrees Celsius. The XRF signal of any which is presumably due to an increase in the amount of receptor-chemical complex formed is then measured. After olanzapine that is bound to 5HT2A, may be measured peri taking this measurement at a first fixed temperature, the tem odically over time to determine the rate at which olanzapine perature is adjusted so that it is different from the first tem binds to 5HT2A. This may be repeated using solutions having perature by at least 2 degrees Celsius. The XRF signal should various concentrations of olanzapine, and at different tem then be measured for the sample at this new temperature. peratures in order to determine the rate of binding at different 0154 Alternatively, the first measurement and second temperatures; this is important for determining the activation measurement can be performed as follows. The receptor and parameters AG, AH, AS, as well as for determining the chemical can be mixed first at one temperature, and next at a binding kinetics that would be expected for a healthy patient different temperature. Then, the chemical-receptor com (37°C.) versus a patient with a mild (38°C.) or severe (>38° plexes that form are isolated (e.g. by spatially separation) so C.) fever. that any chemical that was bound to the receptor remains 0159 For examining a chemical that is a candidate for spatially associated with the receptor. The XRF measure drug discovery according to the present invention, it is pref ments may then be obtained at any temperature desired. erable that the chemical has one or more features that have 0155 The kinetic parameters for the formation of a recep come to be known as “Lipinski’ features, described in C. A. tor-chemical complex may similarly be determined by Lipinski et al., “Experimental And Computational obtaining the baseline XRF signal for at least one receptor as Approaches To Estimate Solubility And Permeability In Drug described above, followed by exposing the receptor to a first Discovery And Development Settings.” Advanced Drug solution of a chemical where the chemical is present in a fixed Delivery Reviews, Vol. 23, pp. 3-25, (1997). These features concentration. The XRF signal of any chemical-receptor include the following: a molecular weight equal to or less than complex is then measured. These steps are then repeated with about 500 daltons; a number of hydrogen bond acceptors that a second solution having a different concentration of the is equal to five or less; a number of hydrogen bond donors that chemical. The contact times between the first solution and the is equal to ten or less; and a log P of five or less where P is the receptor and the second Solution and the receptor are mea ratio of the solubility of the chemical in octanol to the solu sured in order to determine the rate of the binding reaction, bility of the chemical in water. Elaborating on the hydrogen which is related to the amount of complex formed per unit bond acceptor properties of drug candidates, it is preferable time. Preferably the contact times between the first solution that the chemical has the property that the Sum of nitrogen, and the receptor and the second solution and the receptor are oxygen, Sulfur, fluorine, chlorine, bromine, and iodine groups the same. available for hydrogen bonding in the chemical is five or less. 0156 The method may also be practiced using a solution For the purposes of counting hydrogen bond acceptors, each having a concentration gradient; using a concentration gradi of the following is counted as a single hydrogen bonding ent is similar to using two solutions at two different times. The acceptor: difluoromethyl groups, trifluoromethyl groups, reaction kinetics may then be determined, as described in dichloromethyl groups, trichloromethyl groups, dibromethyl Gordon M. Barrow, Physical Chemistry, 5th Ed., McGraw groups, tribromethyl groups, ester groups, amide groups, car Hill, NY, 1988, chapter 18. The activation parameters AG, boxylic acid groups, urea groups, and carbamate groups. Also AH, AS may be determined using the Arrhenius equation for the purposes of counting hydrogen bond acceptor groups, (see, for example: Gordon M. Barrow, Physical Chemistry, the following are not counted as a hydrogen bond acceptor: 5th Ed., McGraw-Hill, NY, 1988, pp. 710-712.) and the nitro groups, nitroso groups, and cyano groups. Eyring equation (see, for example: Gordon M. Barrow, Physi 0160 While the Lipinski features suggest that a molecular cal Chemistry, 5th Ed., McGraw-Hill, NY, 1988, pp. 756 weight of less than about 500 is preferable, Hemmilä (vide 757.) infra) reported that chemicals having a molecular weight of 0157. This aspect of the invention may be demonstrated by 5000 daltons or less cannot be tagged with an optical tag preparing a protein array that includes at least one protein without overly perturbing their binding properties, such as receptor (see, for example: Kukaret al., “Protein Microarrays binding affinities (see I. A. Hemmilä and P. Hurskainen, to Detect Protein-Protein Interactions. Using Red and Green “Novel detection strategies for drug discovery.. Drug Dis Fluorescent Proteins.” Analytical Biochemistry, Vol. 306, pp. covery Today, Vol. 7, pp. S150-S156, (2002)). The present 50-54 (2002), incorporated herein by reference). The baseline invention does not employ tags and thus can be used for sulfur content of the protein spots of the array is determined studying the binding of chemicals that have a molecular using X-ray fluorescence spectroscopy. After determining the weight of 5000 daltons or less. baseline Sulfur content, the protein array is contacted with a 0.161 For a chemical having hydrogen bond donor groups, solution of olanzapine. Afterward, the sulfur content of each it is preferable that the chemical has the property that the sum protein spot is determined again using X-ray fluorescence of hydrogen atoms bound to Sulfur, nitrogen, and oxygen in microscopy. The difference in sulfur content between the two the chemical is ten or less. US 2015/0309021 A1 Oct. 29, 2015

0162 For drug candidates, it is preferable that chemical is the tumor would then be contacted with the protein chip. The soluble in water. The chemical should be dissolved in a fluid binding affinity of each drug would then be measured accord that includes water, preferably with a pH of from about pH 6 ing to the invention, which involves measuring the XRF sig to about pH 8. nals for heavy elements present in the drug-receptor complex, 0163 The chemical may have Zero, one, or more than one Subtracting out the baseline from this measurement, and then chiral center. A chemical sample that contains equal amounts comparing the signals to see which drug exhibits a more of each enantiomer may be used. Once it has been determined selective binding to the tumor versus the healthy tissue and/or using the invention that the chemical binds to a receptor, the which drug binds more strongly to the tumor. chemical may be analyzed to determine which particular 0.168. The efficacy of a drug may be correlated with the enantiomer, if any, is bound to the receptor. For the case of binding affinity of the drug for diseased tissue; more effica more than one enantiomer binding to the receptor, it may be cious drugs tend to bind more strongly to proteins associated determined which is bound more strongly to the receptor. It is with a disease than less efficacious drugs do. Side effects of a important to determine which receptor(s) bind to each enan drug may be correlated with the ability of the drug to bind to tiomer of the chemical in order to identify any potential side healthy tissue. The choice of which drug to prescribe may be effects of a toxic enantiomer. Thalidomide, for example, is a based on the ability of the drug to bind strongly to the diseased chemical that was used to treat morning sickness until it was tissue (or to proteins associated with the diseased tissue) and discovered that a toxic enantiomer of the chemical was to bind weakly, or not at all, to healthy tissue (or to proteins responsible for birth defects. associated with health tissue). A cancer biopsy sample (i.e. 0164. In order to optimize the balance between the speed from a tumor), for example, may be lysed and arrayed on an of analysis versus the sensitivity of the XRF technique, it is analytical protein chip. The chip would then be exposed to a preferable that a sample of receptor, or chemical-receptor variety of chemotherapy drugs (or even cold radiopharma complex, be exposed to the X-ray excitation beam for a period ceutical Surrogates) and the relative binding affinities of the of time from about 100 milliseconds to about 600 seconds. To different drugs to the arrayed biopsy sample would guide the further optimize speed versus sensitivity, it is preferable that course of treatment. Side effects could be quantified and the X-ray excitation beam spends more time focused on a minimized by performing a similar assay using non-cancer pixel that contains receptor and/or receptor-chemical com ous tissue, and selecting the drug based on minimized drug plex than on a pixel that is substantially devoid of receptor or enzyme interactions. This method may also be used with drug receptor-chemical complex. This way, less time is wasted mixtures, which are sometimes referred to as "cocktails'. analyzing parts of a sample that have little useful information. 0169. The invention may be used for participant stratifica 0.165 An X-ray translucent barrier may be used with a tion in clinical trials of proposed drugs. Currently, participant receptor or receptor array in order to minimize evaporation stratification methods are based on DNA chips, where and evaporative cooling of receptors and/or chemicals. genomic profiles are correlated with the variation in partici 0166 Another aspect of the present invention relates to pant response to chemicals (e.g. drug candidates). The corre use for personalized medicine. Personalized medicine is lation of genomic profiles with patient response is sometimes described in “Personalized Prescribing.” The Scientist, referred to as “pharmacogenomics'. Genomic profiles are 1512:10, Jun. 11, 2001, incorporated herein by reference. several steps removed from the proteomic profile of a partici Personalized medicine is important because it is expected to pant in a clinical trial of a drug; genetics and environmental decrease the number of adverse drug reactions, which cur factors affect which proteins are expressed. Drugs regulate rently result in the deaths of about 100,000 hospitalized protein behavior, and so the protein make-up of a particular patients per year, and cause serious side effects in another 2.2 participant and how those proteins interact with the drug(s) in million people. Personalized medicine currently relies on a course of clinical trials will provide the best information for genotyping a patient in order to guide the physician in admin participant stratification. Protein chips have become inexpen istering drugs to the patient. This genotyping focuses on sive and commercially available. single nucleotide polymorphism (SNP) analysis. The cost per 0170 Regarding the use of the invention for participant SNP analysis currently is about 1 dollar per SNP, but with the stratification in a clinical trial of a drug Such as an anti-cancer current poor understanding of pharmacogenomics and the drug, a protein array on a protein chip may be exposed to large number of possible SNPs (thousands), it is believed protein extracts from a cancer patient, as described above. that costs related to SNP analysis must decrease to pennies Protein extracts of the tumor and of healthy tissue would be per SNP before SNP analysis becomes more widely used. The taken from the patient. After exposing the chip to the extracts, analysis provided using the present invention, by contrast, a baseline XRF baseline spectrum would be obtained. Solu allows phenotyping and direct analysis of drug interactions tions of each drug being tested in the trial would then be with protein profiles, thus providing higher quality informa contacted with the protein chip having the bound proteins. tion about the particular patient and disease The binding affinity of each drug would then be measured 0167 Thus, the present invention may be used in person according to the invention, which involves measuring the alized medicine to assist physicians in prescribing drugs for XRF signals of heavy elements present in the drug-receptor their patients. A protein array, Such as an array on a commer complex, Subtracting out the baseline from this measurement, cially available protein chip manufactured by CiphergenTM or and then comparing the signals to see which drug exhibits a ZyomyxTM for example, may be exposed to protein extracts more selective binding to the tumor versus the healthy tissue from a cancer patient. In this case, as in others where there is and/or which drug binds more strongly to the tumor. This a clear difference between diseased tissue and healthy tissue, information may then be correlated to the efficacy, toxicity, extracts from the tumor and from healthy tissue would be and side effects of the drugs as measured in the clinical trials. taken from the patient. After exposing the chip to the extracts, The importance of clinical trial stratification is that if a drug a baseline XRF baseline spectrum would be obtained. Solu can be shown to only have unacceptable side effects for tions of each drug that the doctor is considering for treating people with specific genomic and/or proteomic profiles (i.e. a US 2015/0309021 A1 Oct. 29, 2015 69 strata), the drug may be approved for other strata for which X-ray fluorescence signal that may be correlated with the the drug has acceptable side effects. This process may allow amount of the element in the protein, which provides an drugs that would otherwise be rejected by the Food and Drug estimate of the extent of phosphorylation of the protein. Administration or other regulatory agencies to be accepted (0176) The following EXAMPLES provide an illustration for a more limited use, thereby allowing drug candidates to be of various aspects of this invention. used that would otherwise be rejected. 0171 Another aspect of the invention relates to drug Example 1 manufacturing. The drug manufacturing process is lengthy, typically including a period of time where a new drug is Binding of Ziprasidone to Protein Receptors discovered, another period of time where the drug is devel 0177. A protein array may be prepared as described in G. oped, more time where the drug is tested with animals and MacBeath and S.L. Schreiber, “Printing Proteins. As Microar then humans, and more time where the drug is synthesized on rays For High-Throughput Function Determination. Sci a larger and larger scale. Discovery involves determining ence, vol. 289, pp. 1760-1763, (2000). This protein array may which chemicals are likely to be effective drugs, and is often be exposed to a solution of Ziprasidone, which is the active conducted by measuring the interaction between a chemical ingredient in the drug GeodonTM. Ziprasidone has a molecu and a receptor (sometimes referred to in the art as a "drug gable target”). Development involves in vitro testing to esti lar formula of CHCINOS and the structure shown below. mate the likely side effects and pharmacokinetics of a chemi cal. Animal testing is used to determine whether the drug is safe in animals, and are sometimes used to determine whether a drug is effective in animals. If the drug is deemed suffi ciently safe in animals, it may be administered to human Volunteers in clinical trials to determine human safety and / N N efficacy. If a drug is deemed safe and effective in human Volunteers, it may be mass-produced for use by physicians and patients. All of these steps are necessary. - \ / Sl 0172. The invention may be used to estimate the efficacy C N O of a chemical (analog, drug). Measuring the affinity of one or Ziprasidone more chemicals for a druggable target fulfills the discovery step. 0173 The invention may be used to estimate side effects 0.178 The elements chlorine and sulfur may each be and/or pharmacokinetics by measuring the binding selectiv detected by X-ray fluorescence. After the array is exposed to ity of one or more chemicals for a druggable target versus the Ziprasidone-containing solution, an EDAX Eagle Micro other proteins that may be associated with side effects or probe X-ray fluorescence microscope, for example, could be pharmacokinetics. Animal studies and human studies may used to determine the receptor proteins that bind to the then be performed, followed by mass production of the drug Ziprasidone. The amount of Ziprasidone that is associated using chemical synthesis or biological synthesis. with each receptor may be quantified by X-ray fluorescence. 0.174 Another aspect of the invention relates to its use in 0179 None of the proteins of the array are expected to measuring post-translational modification of proteins. Such contain the element chlorine, so the chlorine X-ray fluores as the phosphorylation or dephosphorylation of one or more cence baseline may be assumed to be Zero (although the proteins. Phosphorylation and desphosphorylation reactions chlorine x-ray fluorescence baseline could be measured). At are described by K. Martin et al. in “Quantitative Analysis Of least Some of the proteins are expected to contain the element Protein Phosphorylation Status And Protein Kinase Activity Sulfur (from Sulfur containing amino acids such as cysteine On Microarrays. Using A Novel Fluorescent Phosphorylation and methionine). Before exposing the array proteins to Sensor Dye.” Proteomics, vol. 3, pp. 1244-1255 (2003). This Ziprasidone, a baseline X-ray fluorescence signal for Sulfur paper describes the need for radioactive labeling and fluores may be obtained, and after exposure to the Ziprasidone, the cent tagging; however, both radioactive labeling and fluores X-ray fluorescence signal due to Sulfur would be measured cent tagging are unnecessary in view of the present invention, again. The difference in the sulfur signals would be used to which can measure the amount of phosphorus directly in an determine whether any receptor-Ziprasidone complex untagged protein using XRF. formed, and measuring the amount of any such complex that 0.175. An example of the use of the invention for measur formed. ing post-translational modifications may include establishing a baseline X-ray fluorescence signal for a heavy element (i.e. Example 2 an element having an atomic number of nine or greater). Such as phosphorus, in a portion of a protein. The portion of protein Binding of Ziprasidone to DNA Receptors is then exposed to reaction conditions such as those described 0180 A DNA array such as an Affymetrix GeneChipTM by K. Martin et al. in “Quantitative Analysis Of Protein Phos may be exposed to a solution that contains Ziprasidone, as phorylation Status And Protein Kinase Activity On Microar described in EXAMPLE 1. After the array is exposed to the rays. Using A Novel Fluorescent Phosphorylation Sensor Ziprasidone-containing Solution, an EDAX Eagle Micro Dye.” Proteomics, Vol. 3, pp. 1244-1255, (2003), which alter probe X-ray fluorescence microscope, for example, could be the amount of phosphorus in the protein. The X-ray fluores used to determine the locations of those receptor DNA that cence signal due to the element would then be remeasured bind to the ziprasidone. The amount of ziprasidone that is using XRF. Subtracting the baseline X-ray fluorescence signal associated with each receptor may be quantified by X-ray from the remeasured X-ray fluorescence signal provides a net fluorescence. US 2015/0309021 A1 Oct. 29, 2015 70

0181. None of the DNA receptors of the array are expected to contain the elements chlorine or sulfur, so the chlorine Olanzapine: X-ray fluorescence baseline and the sulfur X-ray fluorescence CH baseline may be assumed to be zero. After exposure to the / 3 Ziprasidone-containing Solution, the X-ray fluorescence sig nal due to sulfur and/or chlorine would be measured. The s N measured X-ray fluorescence signal(s) may be used to deter mine whether any DNA-ziprasidone complex formed, and NS 2 measuring the amount of any such complex that formed. 0182. The DNA receptors of the array may then be ana lyzed to identify which of the DNA receptor(s) bind to the -( ) ziprasidone and which fail to bind. With this information, I S CH3 which is provided by this EXAMPLE, factors that affect H binding may be determined. Aripiprazole: Example 3 C Binding of DNA Receptors to DNA Probe Molecules (3- N-CH2CH2CH2CH2O N O 0183) A DNA array such as an Affymetrix GeneChipTM may be exposed to a solution of DNA molecules that may or 0187 Olanzapine may be detected and quantified by the may not be complementary to the DNA on the array. If the X-ray fluorescence of its Sulfur atom and aripiprazole may be DNA in solution is not complementary to any of the DNA on detected and quantified by the X-ray fluorescence of its chlo the array, then no binding is expected to occur. If the DNA in rine atoms. solution were complementary to any of the DNA on the array, 0188 After the array is exposed to the solution that con then some binding would be expected to occur. DNA mol tains a mixture of aripiprazole and olanzapine, the array may ecules include the element phosphorus that is detectable be analyzed using an EDAX Eagle Microprobe X-ray fluores using X-ray fluorescence of the phosphorus atoms. After the cence microscope. The protein receptors that bind to olanza array is exposed to the solution of DNA, the array may be pine may be identified by the sulfur X-rays. The amount of analyzed using an EDAX Eagle Microprobe X-ray fluores olanzapine that is associated with each receptor may be quan cence microscope. The DNA receptors of the array that bind tified by X-ray fluorescence. The protein receptors that bind to to the added DNA from the DNA-containing solution may be aripiprazole may be identified by the chlorine X-rays. The identified by the phosphorus X-rays, and the amount of added amount of aripiprazole that is associated with each receptor DNA that is associated with each receptor may be quantified may be quantified by X-ray fluorescence. by the X-ray fluorescence signal. 0189 Preferably, a baseline x-ray fluorescence signal for 0184 Preferably, a baseline x-ray fluorescence signal for chlorine and one for sulfur are measured for each receptor each portion of DNA receptor would be obtained before before exposing the receptors to the Solution. After contacting exposing the receptor array to the DNA containing solution to the array with the solution, the sulfur and chlorine content of determine the phosphorus content of the DNA receptors of each receptor would be measured again. The difference in the the array. After exposing the array to the DNA solution, the measured Sulfur and chlorine X-ray fluorescence signal after difference in the phosphorus X-ray fluorescence signals after contact and before contact with the solution of aripiprazole contact and before contact with the DNA solution may be and olanzapine solution may be quantified and related to the quantified and related to the binding of DNA from the solu binding of aripiprazole and olanzapine to receptors in the tion to various receptors of the array. array. 0185. This DNA receptor array used with the EXAMPLE may be replaced with an array that includes other molecules, Example 5 such as RNA and peptides, to examine the binding of DNA from a solution to these other receptors. RNA may also be Binding of Drugs to Protein Receptors used as a probe molecule. 0190. A protein array may be prepared as described in Example 4 Kukar et al., “Protein Microarrays to Detect Protein-Protein Interactions Using Red and Green Fluorescent Proteins.” Binding of Drugs to Protein Receptors Analytical Biochemistry, vol. 306, pp. 50-54 (2002). This protein array may be exposed to a solution that contains a 0186 A protein array may be prepared as described in mixture olanzapine (the active ingredient in ZyprexaR) and Kukar et al., “Protein Microarrays to Detect Protein-Protein acetylsalicylic acid (the active ingredient in aspirin). Olanza Interactions Using Red and Green Fluorescent Proteins.” pine has the chemical formula C7HNS, and acetylsali Analytical Biochemistry, vol. 306, pp. 50-54 (2002). This cylic acid has the chemical formula CHO. Olanzapine may protein array may be exposed to a solution of olanzapine (the be detected and quantified by the X-ray fluorescence of its active ingredient in ZyprexaR) and aripiprazole (the active sulfur atom. Acetylsalicylic acid, however is more difficult to ingredient in AbilifyTM). Olanzapine has the chemical for both to detect and differentiate using X-ray fluorescence mula C7HNS, and aripiprazole has the chemical formula because it only contains carbon, oxygen and hydrogen, the CHCl2NO. These molecules have the chemical struc X-ray fluorescence signals of which are overwhelmed by the tures shown below. corresponding signals in the protein receptors. US 2015/0309021 A1 Oct. 29, 2015

0191 After the array is exposed to the solution, the array 0198 Preferably, the a baseline X-ray fluorescence signal may be analyzed using an EDAX Eagle Microprobe X-ray for each receptor protein of the array is obtained before expo fluorescence microscope. The protein receptors that bind to Sure to the Solution. After contacting the array with the solu olanzapine in the presence of acetylsalicylic acid may be tion, the Sulfur and chlorine content of each protein receptor identified by the net X-ray fluorescence signal from sulfur. spot may be measured again. The difference in the Sulfur and The amount of olanzapine that is associated with each recep chlorine after contact and before contact with the solution tor may be quantified by X-ray fluorescence. may be quantified and related to the binding of aripiprazole 0.192 Preferably, a baseline X-ray fluorescence measure and olanzapine to the receptors in the array. ment for the protein receptors of the array would be obtained 0199 Antherapeutic index for this patient for olanzapine before exposure to the Solution containing acetylsalicylic and one for aripiprazole may be estimated using the X-ray acid and olanzapine. This analysis may be used to determine fluorescence data. These estimated indices may be used to the sulfur content of the protein spots on the array. After guide the choice of drug to be administered. contacting the array with the solution, the Sulfur content of each protein spot may be measured again. The difference in Example 7 the sulfur after contact and before contact with acetylsalicylic acid and olanzapine solution may be quantified and related to Patient Stratification the binding of olanzapine to various protein receptors in the 0200. A series of protein arrays may be prepared as array when acetylsalicylic acid is present. described in Kukar et al., “Protein Microarrays to Detect 0193 For comparison, the procedure described above for Protein-Protein Interactions. Using Red and Green Fluores this EXAMPLE may be repeated using a solution that cent Proteins.” Analytical Biochemistry, vol. 306, pp. 50-54 includes olanzapine without acetylsalicylic acid in order to (2002). The protein arrays would be prepared using samples determine whether the presence of acetylsalicylic acid has from participants in clinical trials of pharmaceutical com any effect on the binding affinity and binding selectivity of pounds and/or formulations. This is an EXAMPLE for a olanzapine to the protein receptors. After contacting the array study for determining the safety and/or efficacy of olanzap with the Solution containing olanzapine (without acetylsali 1C. cylic acid), the Sulfur content of each protein spot may be 0201 The protein arrays described above would be measured again. The difference in the Sulfur after contact and exposed to a solution that includes olanzapine (C7HNS). before contact with olanzapine solution may be quantified Olanzapine may be detected and quantified by the X-ray fluo and related to the binding of olanzapine to various spots in the rescence of its sulfur atom. array when acetylsalicylic acid is not present. The binding 0202 After the array is exposed to the solution, the array information may be compared to the binding of olanzapine to may be analyzed using an EDAX Eagle Microprobe X-ray various spots in the array when acetylsalicylic acid is present. fluorescence microscope. The protein receptors that bind to (0194 This EXAMPLE illustrates a particularly important olanzapine may be identified by the X-ray fluorescence signal aspect of the present invention, which relates to estimating the due to Sulfur. The amount ofolanzapine that is associated with therapeutic index of a chemical (in this case, olanzapine) and each protein receptor spot may be quantified by X-ray fluo comparing it to an estimated therapeutic index of a "cocktail” CSCCC. i.e. a mixture of two or more chemicals. 0203 Preferably, a baseline X-ray fluorescence signal would be obtained for each receptor before exposure to the Example 6 solution. This analysis may be used to determine the sulfur content of the protein spots on the array. After contacting the Personalized Medicine array with the solution, the sulfur content of each protein spot may be measured again. The difference in the Sulfur content 0.195 A question arises relating to which antipsychotic after contact and before contact with olanzapine Solution may drug, olanzapine or aripiprazole, should be prescribed for a be quantified and related to the binding of olanzapine to patient. A protein array would be prepared as described, for various protein receptors in the array. example, in Kukar et al., “Protein Microarrays to Detect 0204 The binding of olanzapine to various proteins or Protein-Protein Interactions. Using Red and Green Fluores classes of proteins may be then correlated with the results of cent Proteins. Analytical Biochemistry, vol. 306, pp. 50-54 the clinical trial, in order to correlate protein-Olanzapine bind (2002). ing characteristics with safety and/or efficacy of various for 0196. This protein array would be exposed to a solution of mulations of olanzapine-based drugs. a mixture of olanzapine (CHNS) and aripiprazole 0205. It is preferable to use protein arrays rather than DNA (CH,ClNO). Olanzapine may be detected and quanti arrays for patient stratification, because proteins are believed fied by the X-ray fluorescence of its Sulfur atom, and aripipra to control cell functions. However, this method may also be Zole may be detected and quantified by the X-ray fluorescence used with DNA arrays, tissue arrays, cellular arrays, and the of its chlorine atoms. like. 0197) After the array is exposed to the solution, the array 0206. In summary, the present invention provides a would be analyzed using an, EDAX Eagle Microprobe X-ray method for measuring binding selectivities between chemi fluorescence microscope. The receptor proteins that bind to cals and receptors. The invention provides significant advan olanzapine may be identified by their sulfur X-rays. The tages over known methods for measuring binding selectivity. amount of olanzapine that is associated with each receptor Known methods often require either radioactive chemicals, or spot may be quantified by X-ray fluorescence. The receptor chemicals that include a covalently attached label that fluo proteins that bind to aripiprazole may be identified by the resces upon exposure to ultraviolet excitation radiation. Since chlorine X-rays. The amount of aripiprazole that is associated the invention does not require radioactive or chemically with each spot may be quantified by X-ray fluorescence. tagged materials, the problems dealing with the handling of US 2015/0309021 A1 Oct. 29, 2015 72 radioactive materials and the disposal of radioactively con 54. The method of claim 49, wherein the biological recep taminated waste are avoided. Importantly, since the use of tor is selected from the group consisting of a protein, a DNA, artificially tagged materials is not required, there can be no an RNA, a human cell, a plant cell, an animal cell, and a interference from the tag in the evaluation of the binding microorganism, in whole or in part. affinity of the corresponding desired untagged material. Fur 55. The method of claim 49, wherein the chemical to be ther, in contrast to methods that require tags, the method of the measured comprises at least one of fluorine, phosphorous, present invention can be used to evaluate the binding affinity Sulfur, chlorine, bromine, iodine, platinum, copper, Zinc, gal of materials that do not fluoresce while exposed to ultraviolet lium, or gadolinium. radiation. It should be understood that although tagged mate 56. The method of claim 49, wherein in order to reduce rials are not required, they could also be used and this aspect noise, the solid substrate is substantially free of the element to of the invention offers a distinct advantage in that the inven be measured; the substrate is substantially free of radioactive tion can provide a direct comparison of binding affinity of the materials; and for measurement of the element phosphorus, untagged chemical with that of the corresponding tagged the substrate is substantially free of an element selected from Surrogate. This comparison could validate or invalidate the the group consisting of Zirconium, platinum, gold, niobium, assumption that a particular untagged chemical and its tagged mercury, and thallium; for the measurement of the element Surrogate have the same binding affinity to a particular Sub sulfur, the substrate is substantially free of an element Strate. selected from the group consisting of thallium, molybdenum, 0207. The foregoing description of the invention has been Sulfur, lead, bismuth, technetium, and ruthenium; and for presented for purposes of illustration and description and is measurement of the element chlorine, the substrate is sub not intended to be exhaustive or to limit the invention to the stantially free of an element selected from the group consist precise form disclosed, and obviously many modifications ing of technetium, ruthenium, rhodium and palladium. and variations are possible in light of the above teaching. 57. The method of claim 49, wherein the substrate is sub 0208. The embodiment(s) were chosen and described in stantially free of at least one of the elements selected from the order to best explain the principles of the invention and its group consisting of Sulfur, fluorine, phosphorous, chlorine, practical application to thereby enable others skilled in the art bromine, iron, Zinc, magnesium, calcium, titanium, Scan to best utilize the invention in various embodiments and with dium, and platinum. various modifications as are Suited to the particular use con 58. The method of claim 49, wherein the cross section of templated. It is intended that the scope of the invention be the excitation X-ray beam is between 25% and 250% of the defined by the claims appended hereto. area of the receptor-chemical complex. What is claimed is: 1-48. (canceled) 59. The method of claim 49, wherein the chemical and 49. A method for estimating binding of a chemical having chemical-receptor complex are exposed to a buffer which is at least one heavy element of atomic number nine or higher to substantially free of an element present in the chemical to be a biological receptor, comprising: measured and wherein the buffer comprises one or more of providing an X-ray fluorescence signal for a heavy element the chemicals or functional groups selected from the group of in a receptor prior to exposing the receptor to the chemi amine, imine, nitrate anion, nitrite anion, ammonium cation, cal to be tested for binding to the receptor; and iminium cation. exposing the receptor to the chemical and allowing the 60. The method of claim 49, wherein the chemical-receptor chemical to bind to the receptor to form a chemical complex is exposed to a solution prior to measurement, the receptor complex; Solution comprising a matrix modifier. measuring the X-ray fluorescence signals due to the heavy 61. The method of claim 49, wherein the X-ray fluorescence element in the chemical-receptor complex; is measured using a chromium, palladium or silver X-ray wherein the X-ray fluorescence measurements are per SOUC. formed using a solid Substrate having a thickness of less 62. The method of claim 49, wherein the X-ray fluorescence than 500 microns. is measured using a rhodium source. 50. The method of claim 49, further comprising subtracting 63. The method of claim 49, wherein the X-ray fluorescence the X-ray fluorescence signal of the receptor prior to exposing is measured using a molybdenum source. the receptor to the chemical to be tested for binding to the 64. The method of claim 49, wherein the X-ray fluorescence receptor from the measured X-ray fluorescence signal of the is measured using an X-ray Sources having a K-alpha or L-al chemical-receptor complex to quantify the binding of the pha line at or above 2.838 KeV and less than 9.441 KeV. chemical to the receptor. 65. The method of claim 49, wherein the chemical-receptor 51. The method of claim 49, further comprising standard complex is deposited on the substrate in volumes between 1 izing the binding of the chemical to the receptor by dividing nanoliter and 50 microliters. the binding of the chemical to the receptor by the amount of receptor to obtain a standardized binding of the chemical to 66. The method of claim 49, wherein the chemical-receptor the receptor. complex is concentrated on the Substrate prior to measure 52. The method of claim 49, wherein the X-ray fluorescence ment. is provided using a polychromatic X-ray source. 67. The method of claim 49, wherein the sample contains 53. The method of claim 49, wherein the solid substrate between ten nanograms and one microgram of the heavy comprises an element selected from the group consisting of element to be measured. beryllium, carbon, silicon, aluminum, iron, cobalt and gold. k k k k k