US 20150267245A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0267245 A1 Hogan (43) Pub. Date: Sep. 24, 2015

(54) PRESERVATION OF BIOLOGICAL (52) U.S. Cl. MATERIALS IN NON-AQUEOUS FLUID CPC ...... CI2O I/6806 (2013.01); C12O I/686 MEDIA (2013.01) (71) Applicant: GenTegra, LLC, Pleasanton, CA (US) (57) ABSTRACT (72) Inventor: Michael Hogan, Tucson, AZ (US) The invention provides compositions and methods for pre (73) Assignee: GenTegra, LLC, Pleasanton, CA (US) serving a biological material—Such as a protein, a nucleic acid or a biological sample, or any combination thereof in a (21) Appl. No.: 14/213,066 Substantially water-free, nonionic or ionic organic solvent. Improved preservation, including for example the stability (22) Filed: Mar 14, 2014 and/or the solubility of the biological material in the substan tially water-free fluid medium, is achieved with compositions Related U.S. Application Data comprising one or more Substances (e.g., an antioxidant) (60) Provisional application No. 61/786,171, filed on Mar. described in the disclosure, and/or a metal salt. The biological 14, 2013. material is soluble and stable, and retains its function and activity, when it is preserved in the substantially water-free Publication Classification fluid medium at ambient temperature or higher for extended periods of time. Therefore, the composition comprising the (51) Int. Cl. biological material does not need to be refrigerated or frozen CI2O I/68 (2006.01) during shipping or storage. Patent Application Publication Sep. 24, 2015 Sheet 1 of 7 US 2015/0267245 A1

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PRESERVATION OF BIOLOGICAL 0010. It is another object, feature, and/or advantage of the MATERIALS IN NON-AQUEOUS FLUID present invention to provide compositions for preservation of MEDIA biological materials, wherein the biological materials are soluble and stable in the substantially water-free fluid media CROSS-REFERENCE TO RELATED at ambient temperature or higher for extended periods of APPLICATIONS time, and thus do not need to be refrigerated or frozen during shipping or storage. 0001. This application claims priority under 35 U.S.C. 0011. In one aspect, the present invention provides meth S119 of a provisional application Ser. No. 61/786,171 filed ods and compositions for preservation of biological materi Mar. 14, 2013, which is hereby incorporated by reference in als. Such as proteins, nucleic acids and biological samples, in its entirety. substantially water-free fluid media. The biological materials are soluble and stable in the substantially water-free fluid GRANT REFERENCE media at ambient temperature or higher for extended periods 0002 This invention was made with government support of time, and thus do not need to be refrigerated or frozen under Contract No. HR0011-12-C-0005 awarded by the during shipping or storage. The biological materials retain Defense Advanced Research Projects Agency (DARPA). The their structural integrity, function and activity after preserva government has certain rights in the invention. tion in the substantially water-free fluid media at ambient temperature or higher for extended periods of time. Because FIELD OF THE INVENTION the biological materials are preserved in a fluid medium, they may not need to be re-dissolved for use in fluid-phase reac 0003. The present invention relates to preservation of bio tions or assays, including nucleic acid amplification reactions logical materials, such as proteins, nucleic acids and biologi based on polymerase chain reaction (PCR) and analytical and cal samples, in Substantially water-free fluid media. diagnostic assays, such as immunoassays. 0012. In one aspect, the present invention provides com BACKGROUND OF THE INVENTION positions comprising a biological material in a Substantially 0004 Biological materials, such as proteins and nucleic water-free fluid medium, wherein the fluid medium com acids, may in certain instances be freeze dried (lyophilized) to prises a non-ionic organic solvent (e.g., an alcohol solvent) or enhance their stability in the absence of refrigeration. Freeze an ionic organic solvent comprising an organic salt and an drying comprises freezing of an aqueous mixture containing organic hydrogen bond donor. To enhance, e.g., the Stability a biological material and removal of water via Sublimation. and/or the solubility of the biological material in the substan Biological materials can Suffer denaturation including for tially water-free fluid medium, the compositions can further example, partial denaturation—as a result of freeze-drying, comprise a metal salt, and/or one or more Substances selected for example, as a result of the freezing step and/or the Subli from the group consisting of reducing agents, antioxidants, mation step. Thus, there is a need in the art for methods, free radical scavengers, oxygen radical scavengers, hydroxyl compositions, and systems for stable preservation of biologi radical scavengers, singlet oxygen quenchers, hydroperoX ide-removing agents, protease inhibitors, nuclease inhibitors, cal materials that prevents denaturation. ribonuclease (RNase) inhibitors, deoxyribonuclease (DNase) inhibitors, metal chelators, preservatives, anti-microbials, SUMMARY OF THE INVENTION buffers (or buffering agents), detergents, and chaotropes. 0005. It is therefore a primary object, feature, and/or 0013. In another aspect, the present invention provides advantage of the present invention to improve on or overcome methods of preserving a biological material, comprising mix the deficiencies in the art. ing an aqueous mixture comprising a biological material with 0006. It is another object, feature, and/or advantage of the a non-ionic organic solvent (e.g., an alcohol Solvent) or an present invention to provide methods for preservation of bio ionic organic solvent comprising an organic salt and an logical materials, such as proteins, nucleic acids and biologi organic hydrogen bond donor to produce an aqueous organic cal samples, in Substantially water-free fluid media. mixture, and removing water from the aqueous organic mix 0007. It is another object, feature, and/or advantage of the ture to produce a substantially water-free fluid medium com present invention to provide methods for preservation of bio prising the biological material and the non-ionic or ionic logical materials, wherein the biological materials are soluble organic solvent. The fluid medium can further comprise a and stable in the substantially water-free fluid media at ambi metal salt and/or one or more Substances as described herein. ent temperature or higher for extended periods of time, and 0014. In another aspect, the present invention provides thus do not need to be refrigerated or frozen during shipping containers and kits containing compositions that comprise or Storage. biological materials in substantially water-free fluid media. 0008. It is another object, feature, and/or advantage of the 0015 While multiple embodiments are disclosed, still present invention to provide methods of preserving biological other embodiments of the present invention will become material in a non-aqueous fluid media, including, for apparent to those skilled in the art from the following detailed example, a non-ionic organic solvent such as an alcohol Sol description, which shows and describes illustrative embodi vent or an ionic organic solvent comprising an organic salt ments of the invention. Accordingly, the drawings and and an organic hydrogen bond donor to produce an aqueous detailed description are to be regarded as illustrative in nature organic mixture. and not restrictive. 0009. It is another object, feature, and/or advantage of the present invention to provide compositions for preservation of BRIEF DESCRIPTION OF THE DRAWINGS biological materials, such as proteins, nucleic acids and bio 0016. The following figures are included to illustrate cer logical samples, in Substantially water-free fluid media. tain aspects of the present invention, and should not be viewed US 2015/0267245 A1 Sep. 24, 2015

as exclusive embodiments. The subject matter disclosed is applies to each one of the numerical values in that series of capable of considerable modifications, alterations, combina numerical values or in that series of ranges of numerical tions, and equivalents in form and function, as will occur to values. In certain embodiments, the term “about' or “approxi those skilled in the art and having the benefit of this disclo mately” means within 10% or 5% of the specified value. SUC. (0027. Whenever the term “at least” or “greater than' pre 0017 FIG. 1 shows electrophoresis results of reverse tran cedes the first numerical value in a series of two or more scription PCR (RT-PCR) for analysis of human 18S riboso numerical values, the term “at least or “greater than applies mal RNA (rRNA) after preservation of RT-PCR reagents in to each one of the numerical values in that series of numerical glycerol, with addition of no Sucrose or varying amounts of values. Sucrose, at ambient temperature for varying periods of time. 0028. Whenever the term “no more than or “less than 0018 FIGS. 2 (A-B) shows electropherograms of multi precedes the first numerical value in a series of two or more plex PCR for analysis of all 13 short tandem repeat (STR) loci numerical values, the term “no more than' or "less than utilized in the CODIS forensic database, as well as Penta D, applies to each one of the numerical values in that series of Penta E and amelogenin, after preservation of PCR reagents numerical values. in glycerol, with addition of no Sucrose or varying amounts of Sucrose, at ambient temperature at time 0. 0029 Percentage of a substance by mass is understood to 0019 FIG.3 (A-B) shows electropherograms of multiplex refer to the amount of a component or portion of a combined PCR for analysis of all 13 short tandem repeat (STR) loci or entire composition in reference to the amount represented utilized in the CODIS forensic database, as well as Penta D, by the combined or entire composition. For example, if a fluid Penta E and amelogenin, after preservation of PCR reagents is 10% water by mass, it is understood that the water repre in glycerol, with addition of no Sucrose or varying amounts of sents 10% of the mass represented by the entire fluid, includ Sucrose, at ambient temperature after 7 days. ing the water. 0020 FIG. 4 shows mixtures containing human serum 0030. In some embodiments, the term "ambient tempera Solids and varying amounts of glycerol and Sucrose after ture' or “room temperature” refers to a temperature range removal of water under reduced pressure. from about 18°C. to about 27°C., or from about 20° C. to 0021 FIG.5 (A-B) shows a electrophoresis results of RT about 25°C., or from about 22°C. to about 25°C. In other PCR analysis of human 18S rRNA performed at time-0 (A) embodiments, the term "ambient temperature' or “room tem or about 7 days after (B) sucrose and glycerol or 1,3-pro perature” refers to a temperature of about 18°C., 19°C., 20° panediol, and optionally mineral oil or a detergent, had been C., 21°C., 22°C., 23° C., 24°C., 25°C., 26° C. or 27°C. In added to a master mix of RT-PCR reagents. certain embodiments, the term "ambient temperature' or “room temperature' refers to a temperature of about 22°C., DETAILED DESCRIPTION OF THE INVENTION 23° C. 24°C. or 25° C. 0022 While various embodiments of the present disclo 0031. The term “halide' refers to fluoride, chloride, bro sure are described herein, it will be obvious to those skilled in mide and iodide. the art that such embodiments are provided by way of 0032. The terms “biological reaction' and “biochemical example only. Numerous modifications and changes to, and reaction” are used interchangeably herein unless expressly variations and substitutions of the embodiments described indicated otherwise. herein will be apparent to those skilled in the art without 0033. In one embodiment, the invention provides for pres departing from the disclosure. It is understood that various ervation of biological materials in substantially water-free alternatives to the embodiments described herein may be fluid media. The term “biological materials' refers to natu employed in practicing the disclosure. It is further understood rally occurring molecules and Substances such as proteins, that every embodiment of the disclosure may optionally be nucleic acids and biological samples, or man-made equiva combined with any one or more of the other embodiments lents or homologues thereof. Examples of polypeptide bio described herein which are consistent with that embodiment. logical materials include, but are not limited to, enzyme that 0023 Headings are included herein for reference and to mediate a nucleic acid reaction, polypeptides that regulate an aid in locating certain sections. Headings are not intended to enzyme, antibodies, polypeptide ligands of antibodies, limit the scope of the embodiments and concepts described in polypeptide aptamers, proteins or enzymes useful for detec the sections under those headings, and those embodiments tion, toxins, hormones, cytokines, polypeptide therapeutics, and concepts may have applicability in other sections or vaccines, and derivatives of any of these. Examples of throughout the entire disclosure. nucleic acid biological materials include, but are not limited 0024 All patent literature and all non-patent literature to polynucleotides used in a nucleic acid reaction, catalytic cited herein are incorporated herein by reference in their polynucleotides, or polynucleotides that binds specifically to entirety to the same extent as if each patent literature or a target ligand, and derivatives of any of these. Examples of non-patent literature were specifically and individually indi biological sample biological materials include, but are not cated to be incorporated herein by reference in its entirety. limited to, biological fluids, biological Suspensions, fluid 0025. The term “exemplary” as used herein means “serv aspirates, blood, plasma, serum, lymph, cerebrospinal fluid, ing as an example, instance, or illustration'. Any embodiment gastric fluid, bile, perspiration, ocular fluid, tears, oral fluid, characterized herein as “exemplary' is not necessarily to be sputum, saliva, buccal samples, tonsil samples, nasal construed as preferred or advantageous over other embodi samples, mucus, nasopharyngeal samples, semen, urine, mentS. vaginal samples, cervical samples, rectal samples, fecal 0026. Whenever the term “about” or “approximately pre samples, wound or purulent samples, hair, tissue, tissue cedes the first numerical value in a series of two or more homogenates, cells, cellular lysate, tissue or cell biopsy, skin numerical values or in a series of two or more ranges of cells, tumor or cancer cells, microbes, pathogens, bacteria, numerical values, the term “about' or “approximately fungi, protozoa, and viruses. US 2015/0267245 A1 Sep. 24, 2015

0034. A biological material can be transferred from an water-free, non-ionic organic solvent (e.g., an alcohol Sol aqueous medium to a substantially water-free fluid medium Vent). In certain embodiments, such a composition comprises without passing through an intermediate solid State (e.g., a polypeptide, a polynucleotide or a biological sample, or any without freezing of the aqueous medium or an aqueous combination thereof, and at least one alcohol solvent. In one organic medium), by mixing of an aqueous mixture compris aspect the polypeptide may be, for example, an enzyme that ing the biological material with a non-ionic organic solvent mediates a nucleic acid reaction, a polypeptide that regulates (e.g., an alcohol Solvent) or an ionic organic solvent and an enzyme, an antibody, a polypeptide ligand of an antibody, removal of water from the resulting aqueous organic mixture. a polypeptide aptamer, a protein or enzyme useful for detec Removal of water may be achieved using any number of tion, a toxin, a hormone, a cytokine, a polypeptide therapeutic Solvent removal techniques known in the art, including, for or a vaccine, or a derivative thereof or any combination example, distillation and evaporation. In a preferred embodi thereof. In another aspect the polynucleotide may be, for ment, water removal comprises evaporation. Evaporation example, a polynucleotide used in a nucleic acid reaction, a process and techniques include, for example, open-dish or catalytic polynucleotide, or a polynucleotide that binds spe open-container evaporation, reduced-pressure evaporation, cifically to a target ligand, or a derivative thereof or any and rotary evaporation. combination thereof. In another aspect the biological sample 0035) To enhance, e.g., the stability and/or the solubility of may be, for example, a biological fluid, a biological Suspen the biological material in the substantially water-free fluid Sion, a fluid aspirate, blood, plasma, serum, lymph, cere medium, the fluid medium can comprise a metal salt, and/or broSpinal fluid, gastric fluid, bile, perspiration, ocular fluid, one or more substances selected from the group consisting of tears, oral fluid, sputum, saliva, a buccal sample, a tonsil reducing agents, antioxidants, free radical scavengers, oxy sample, a nasal sample, mucus, a nasopharyngeal sample, gen radical scavengers, hydroxyl radical scavengers, singlet semen, urine, a vaginal sample, a cervical sample, a rectal oxygen quenchers, hydroperoxide-removing agents, protease sample, a fecal sample, a wound or purulent sample, hair, a inhibitors, nuclease inhibitors, ribonuclease (RNase) inhibi tissue, a tissue homogenate, cells, a cellular lysate, a tissue or tors, deoxyribonuclease (DNase) inhibitors, metal chelators, cell biopsy, skin cells, tumor or cancer cells, a microbe, a preservatives, anti-microbials, buffers (or buffering agents), pathogen, a bacterium, a fungus, a protozoan or a virus, or any detergents, and chaotropes. combination thereof. 0036. The biological materials are soluble and stable in the 0039. In another aspect, the alcohol solvent may be substantially water-free fluid media at ambient temperature selected from the group consisting of linear and branched or higher for extended periods of time, and thus do not need to C2-C6 acyclic alcohols having one or more hydroxyl groups be refrigerated or frozen during shipping or storage. In addi and C3-C6 cyclic alcohols having one or more hydroxyl tion, the biological materials retain their structural integrity, groups and three to six ring carbon atoms. The acyclic alco function and activity after preservation in the substantially hols and cyclic alcohols optionally comprise one or more water-free fluid media at ambient temperature or higher for halide atoms. In a preferred embodiment, the composition is extended periods of time. The biological materials retain their in a fluid state and is substantially free of water. structural integrity, function and activity even though alcohol 0040. In some embodiments, the at least one alcohol sol Solvents (including polyol solvents) and ionic organic Sol vent in the composition comprises no more than about 20%, vents (including deep eutectic solvents) comprising an 15%, 10%, 5%, 4%. 3%, 2%, 1% or 0.5% water by mass, organic salt and an organic hydrogen bond donor can dena relative to the combined mass of water and the at least one ture biological materials such as proteins (including alcohol Solvent, e.g., after storage of the composition in a enzymes) and nucleic acids (including double-stranded closed container at a temperature from ambient temperature DNA). Furthermore, the biological materials retain their to about 40° C. for at least about 1 day, 3 days, 1 week, 2 structural integrity, function and activity even though removal weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months, 1 of water from an aqueous organic mixture comprising a bio year, 1.5 years or 2 years. The closable or closed container logical material (e.g., a protein, such as an enzyme) and a salt may be, for example, a capped tube, vial or well. In certain (e.g., an inorganic salt, Such as Sodium chloride) can resultin, embodiments, the at least one alcohol solvent in the compo e.g., at least a 5-fold or 10-fold greater concentration of the sition comprises no more than about 10%, 5% or 1% water by salt in the substantially water-free fluid medium, which high mass after storage of the composition in a closed container at salt concentration may be expected to be deleterious to the a temperature from ambient temperature to about 40°C. for at structure, function and/or activity of the biological material. least about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 0037. In addition to avoiding denaturation that can result 2 months, 3 months, 6 months, 1 year, 1.5 years or 2 years. In from freeze-drying of biological materials, preservation of Some embodiments, the at least one alcohol solvent in the biological materials in substantially water-free fluid media composition comprises no more than about 10%, 5% or 1% can facilitate handling of the biological materials. Unlike water by mass relative to the combined mass of water and the lyophilized biological materials, because the biological mate at least one alcohol solvent after storage of the composition in rials of the present disclosure are preserved in a fluid medium, a closed container at ambient temperature for at least about 3 they may not need to be re-dissolved for use in fluid-phase months or 6 months. reactions or assays, including nucleic acid amplification reac tions based on PCR and analytical and diagnostic assays, Such 0041. In further embodiments, the at least one alcohol as immunoassays. Solvent is Substantially soluble in water—for example, at least about 50%, 60%, 70%, 80%, 90%, 95% or 99% of the at Compositions Comprising a Biological Material in an least one alcohol solvent by mass or volume is soluble in Anhydrous, Non-Ionic Organic Solvent water. In certain embodiments, at least about 90%, 95% or 99% of the at least one alcohol solvent by mass or volume is 0038. Some embodiments of the disclosure relate to com soluble in water. In an embodiment, the at least one alcohol positions comprising a biological material in a Substantially solvent is miscible with water. Solubility of the at least one US 2015/0267245 A1 Sep. 24, 2015

alcohol solvent in water promotes transfer of a biological glycerol, 1.2-butanediol. 1,3-butanediol, 1,4-butanediol. 2,3- material from an aqueous medium to the at least one alcohol butanediol. 1,2,4-butanetriol or 1.5-pentanediol, or any com solvent. bination thereof. In certain embodiments, the at least one 0042. In yet further embodiments, the at least one alcohol alcohol solvent comprises ethylene glycol, 1,3-propanediol. Solvent has a boiling point Substantially greater than that of glycerol or 1.2-butanediol, or any combination thereof. water—e.g., a boiling point at least or greater than about 110° 0047. In some embodiments, the C3-C6 cyclic alcohols C., 125° C., 150° C., 175°C., 200° C. or 250° C. at a pressure having one or more hydroxyl groups and optionally compris of about 1 atmosphere (atm). In certain embodiments, the at ing one or more halide atoms are C4 or C5 cyclic alcohols least one alcohol solvent has a boiling point at least or greater having one or more hydroxyl groups and four or five ring than about 150° C., 175° C. or 200° C. at a pressure of about carbon atoms and optionally comprising one or more halide 1 atm. The at least one alcohol Solvent having a boiling point atoms. In certain embodiments, the at least one alcohol Sol greater than the boiling point of water allows for an aqueous vent comprises cyclobutanol or cyclopentanol, or both. mixture comprising a biological material to be mixed with at 0048. In some embodiments, the at least one alcohol sol least one alcohol solvent and for water to be selectively vent comprises two or more alcohol Solvents. In certain removed (e.g., by evaporation) from the resulting aqueous embodiments, the at least one alcohol solvent comprises two organic mixture without Substantial loss of the at least one or more alcohol solvents selected from the group consisting alcohol solvent. of ethylene glycol, 1.2-propanediol. 1,3-propanediol, glyc 0043. In additional embodiments, the at least one alcohol erol, 1,2-butanediol, 1,3-butanediol, 1,4-butanediol, 2,3-bu Solvent may have a dynamic (or absolute) viscosity of no tanediol. 1,2,4-butanetriol, and 1.5-pentanediol. In additional more than about 1500, 1000, 500, 400, 300, 200, 100, 50 or 25 embodiments, the at least one alcohol solvent comprises: (a) centipoise (cP) or mPa is at ambient temperature. In certain ethylene glycol and glycerol; or (b) ethylene glycol and 1.2- embodiments, the at least one alcohol solvent has a dynamic butanediol; or (c) glycerol and 1.2-butanediol. In further (or absolute) viscosity of no more than about 1000, 500, 200, embodiments, the at least one alcohol Solvent comprises glyc 100 or 50 cp or mPa's at ambient temperature. A lower erol and an additional alcohol solvent that can be any alcohol dynamic (or absolute) viscosity of the at least one alcohol solvent described herein. In certain embodiments, the at least solvent allows for more facile handling of the composition one alcohol solvent comprises glycerol and an additional comprising the biological material and the at least one alcohol alcohol solvent selected from the group consisting of ethyl Solvent (e.g., using a pipette or other means of transferring the ene glycol. 1.2-propanediol. 1,3-propanediol. 1.2-butanediol. fluid composition). 1,3-butanediol, 1,4-butanediol, 2,3-butanediol, 1,2,3-butan 0044. In some embodiments, the linear and branched etriol. 1,2,4-butanetriol, 1.2-pentanediol. 1,3-pentanediol. C2-C6 acyclic alcohols having one or more hydroxyl groups 1,4-pentanediol. 1.5-pentanediol. 2.3-pentanediol. 2.4-pen and optionally comprising one or more halide atoms are lin tanediol. 1.2.3-pentanetriol, 1.2.4-pentanetriol, 1.2.5-pentan ear and branched C2-C5 acyclic alcohols having one etriol, 1.3.4-pentanetriol, 1.3.5-pentanetriol, 2.3.4-pentan hydroxyl group and optionally comprising one or more halide etriol, 1.2-hexanediol, 1,3-hexanediol, 1,4-hexanediol, 1.5- atoms. Non-limiting examples of linear and branched C2-C5 hexanediol. 2,3-hexanediol. 2.4-hexanediol. 2.5-hexanediol. acyclic alcohols having one hydroxyl group and optionally 3,4-hexanediol, di(ethylene glycol), and tri(ethylene glycol). comprising one or more halide atoms include 2-chloroetha 0049. In some embodiments, the composition comprises nol. 2,2-dichloroethanol, 1-butanol, 1-pentanol, 2-methylbu an enzyme. In certain embodiments, the enzyme mediates a tan-1-ol. 3-methylbutan-1-ol. 2,2-dimethylpropan-1-ol, pen nucleic acid reaction. In some embodiments, the enzyme that tanol, 3-methylbutan-2-ol, and 3-pentanol. mediates a nucleic acid reaction comprises a topoisomerase, 0045. In other embodiments, the linear and branched a helicase, a DNA polymerase, a reverse transcriptase, an C2-C6 acyclic alcohols are linear and branched C2-C6 acy RNA polymerase, a DNA ligase, an RNA ligase, a DNA clic alcohols having two or more (e.g., two or three) hydroxyl repair enzyme, an RNA repair enzyme, an endonuclease, an groups and optionally comprising one or more halide atoms. exonuclease, a deoxyribonuclease (DNase), a ribonuclease Non-limiting examples of linear and branched C2-C6 acyclic (RNase), a transposase, a restriction enzyme or a nicking alcohols having two or more (e.g., two or three) hydroxyl enzyme, or any combination thereof. In further embodiments, groups and optionally comprising one or more halide atoms the enzyme that mediates a nucleic acid reaction comprises a include 12-ethanediol (ethylene glycol), 1,2-propanediol DNA polymerase, a reverse transcriptase or an RNA poly (propylene glycol), 1,3-propanediol. 1.2.3-propanetriol merase, or any combination thereof. In certain embodiments, (glycerol), 1.2-butanediol. 1,3-butanediol, 1,4-butanediol. the enzyme that mediates a nucleic acid reaction comprises a 2,3-butanediol. 1,2,3-butanetriol, 1,2,4-butanetriol, 1.2-pen DNA polymerase (e.g., a heat-stable DNA polymerase, such tanediol. 1,3-pentanediol. 1.4-pentanediol. 1.5-pentanediol. as a Taq polymerase) used in PCR or a reverse transcriptase 2,3-pentanediol. 2.4-pentanediol. 1.2.3-pentanetriol. 1,2,4- used in PCR, or both. pentanetriol, 1.2.5-pentanetriol, 1.3.4-pentanetriol, 1.3.5- 0050 Polymerase chain reaction (PCR) includes standard pentanetriol, 2.3.4-pentanetriol, 1.2-hexanediol. 1.3-hex PCR and variations thereof, such as allele-specific PCR, anediol. 1.4-hexanediol. 1.5-hexanediol. 2.3-hexanediol. 2,4- assembly PCR, asymmetric PCR, dial-out PCR, hot-start hexanediol, 2.5-hexanediol, 3,4-hexanediol, di(ethylene PCR, intersequence-specific PCR, inverse PCR, isothermal glycol), and tri(ethylene glycol). PCR (e.g., helicase-dependent amplification and PAN-AC), 0046. In certain embodiments, the linear and branched ligation-mediated PCR, methylation-specific PCR, mini C2-C6 acyclic alcohols are linear and branched C2-C5 acy primer PCR, multiplex ligation-dependent probe amplifica clic alcohols having two or more (e.g., two or three) hydroxyl tion, multiplex PCR, nested PCR, overlap-extension PCR, groups and optionally comprising one or more halide atoms. picotiter PCR, quantitative PCR, real-time PCR, restriction In some embodiments, the at least one alcohol solvent com fragment length polymorphism PCR, reverse transcription prises ethylene glycol, 1.2-propanediol. 1,3-propanediol. PCR (RT-PCR), single-cell PCR, solid-phase PCR (e.g., stan US 2015/0267245 A1 Sep. 24, 2015 dard solid-phase PCR, enhanced solid-phase PCR, bridge one pair of forward and reverse primers comprises 16 differ PCR, and polony amplification), thermal asymmetric inter ent pairs of forward and reverse primers for amplifying all 13 laced PCR, touchdown (step-down) PCR, and universal fast CODIS STR loci, Penta D, Penta E and amelogenin. In some walking PCR. embodiments, each of the 16 different pairs of forward and 0051. In further embodiments, the composition comprises reverse primers is labeled with a dye (e.g., a fluorescent dye), a polynucleotide used in a nucleic acid reaction. In some which may be the same as or different from the dyes used to embodiments, the polynucleotide comprises at least one label the other pairs of forward and reverse primers (e.g., primer used in PCR or reverse transcription. In certain three, four or more spectrally resolvable fluorescent dyes can embodiments, the at least one primer used in PCR or reverse be used to label the 16 different pairs of forward and reverse transcription comprises at least one pair of a forward primer primers). In certain embodiments, the DNA polymerase com and a reverse primer for amplifying at least one nucleic acid prises a DNA polymerase that is stable at elevated tempera (e.g., genetic) locus, or at least one reverse transcription ture (e.g., at about 60° C., 70° C., 80° C., 90° C. or higher), primer for reverse transcribing at least one polyribonucle Such as a Taq polymerase. In further embodiments, the otide, or both. In some embodiments, the at least one pair of reagents for performing PCR further comprise deoxyribo forward primer and reverse primer is labeled with a dye (e.g., nucleotide triphosphates. In additional embodiments, the a fluorescent dye) or the at least one reverse transcription reagents for performing PCR further comprise a buffer or a primer is labeled with a dye (e.g., a fluorescent dye), or both. metal salt (e.g., an M'' or M" salt, such as magnesium A reverse transcription primer refers to a polynucleotide chloride), or both. primer used for reverse transcribing at least one polyribo 0054. In some embodiments, the composition comprises nucleotide to produce at least one poly deoxyribonucleotide the DNA polymerase, the at least one pair of forward and complementary to the at least one polyribonucleotide. reverse primers, deoxyribonucleotide triphosphates, and 0052. In some embodiments, the composition comprises a optionally a buffer and/or a metal salt (e.g., an M' or M" plurality of different pairs of forward and reverse primers for salt, Such as magnesium chloride) for performing PCR. In amplifying a plurality of different short tandem repeat (STR) other embodiments, the composition comprises the DNA loci utilized in a forensic database, such as the Combined polymerase and no primer, where the composition can further DNA Index System (CODIS) recommended by the Federal comprise deoxyribonucleotide triphosphates and optionally a Bureau of Investigation (FBI). CODIS presently utilizes 13 buffer and/or a metal salt (e.g., an M' or M" salt, such as STR loci dubbed CSF1PO, D3S1358, D5S818, D7S820, magnesium chloride). In yet other embodiments, the compo D8S1179, D13S317, D16S539, D18S51, D21S11, FGA, sition comprises the at least one pair of forward and reverse TH01, TPDX and v WA. In certain embodiments, the compo primers and no DNA polymerase, where the composition can sition comprises at least 5 or at least 10, or 13, different pairs further comprise deoxyribonucleotide triphosphates and of forward and reverse primers for amplifying at least 5 or at optionally a buffer and/or a metal salt (e.g., an M' or M" least 10, or all 13, CODIS STR loci. In further embodiments, salt, such as magnesium chloride). A kit containing reagents the composition further comprises at least one pair of forward for performing PCR can contain, for example: a composition and reverse primers for amplifying at least one other STR comprising the DNA polymerase, the at least one pair of locus useful for human identification, such as Penta D and forward and reverse primers, deoxyribonucleotide triphos Penta E. In additional embodiments, the composition further phates, and optionally a buffer and/or a metal salt; or (i) a comprises at least one pair of forward and reverse primers for composition comprising the DNA polymerase and no primer, amplifying at least one nucleic acid (e.g., genetic) locus use which can further comprise deoxyribonucleotide triphos ful for sex determination, such as amelogenin (AMEL). In phates and optionally a buffer and/or a metal salt; and (ii) a certain embodiments, the composition comprises 16 different separate composition comprising the at least one pair of for pairs of forward and reverse primers for amplifying all 13 ward and reverse primers and no DNA polymerase, which can CODIS STR loci, Penta D, Penta E and amelogenin. In some further comprise deoxyribonucleotide triphosphates and embodiments, each of the 16 different pairs of forward and optionally a buffer and/or a metal salt. reverse primers is labeled with a dye (e.g., a fluorescent dye), 0055. In some embodiments, the composition comprises which may be the same as or different from the dyes used to one or more PCR reagents for amplifying at least one nucleic label the other pairs of forward and reverse primers (e.g., acid (e.g., genetic) sequence of the DNA of a microbe or a three, four or more spectrally resolvable fluorescent dyes can pathogen, such as the DNA of any bacterium described be used to label the 16 different pairs of forward and reverse herein, the DNA of any fungus described herein, and the DNA primers). of any DNA virus described herein. A kit can contain a com 0053. In additional embodiments, the composition com position comprising all of the PCR reagents for amplifying at prises one or more reagents for performing PCR, wherein the least one nucleic acid (e.g., genetic) sequence of the DNA of reagents for performing PCR comprise a DNA polymerase a microbe or a pathogen, or two or more compositions that in and at least one pair of a forward primer and a reverse primer total comprise all of the PCR reagents. for amplifying at least one nucleic acid (e.g., genetic) locus, 0056. In other embodiments, the composition comprises and the at least one pair of forward primer and reverse primer one or more reagents for performing reverse transcription, optionally is labeled with a dye (e.g., a fluorescent dye). In wherein the reagents for performing reverse transcription Some embodiments, the at least one pair of forward and comprise a reverse transcriptase and at least one reverse tran reverse primers comprises at least 5 or 10 different pairs of Scription primer for reverse transcribing at least one polyri forward and reverse primers for amplifying at least 5 or 10 bonucleotide, and the at least one reverse transcription primer different STR loci utilized in a forensic database (e.g., optionally is labeled with a dye (e.g., a fluorescent dye). In CODIS), wherein each of the at least 5 or 10 different pairs of further embodiments, the reagents for performing reverse forward and reverse primers optionally is labeled with a dye transcription further comprise deoxyribonucleotide triphos (e.g., a fluorescent dye). In certain embodiments, the at least phates. In additional embodiments, the reagents for perform US 2015/0267245 A1 Sep. 24, 2015 ing reverse transcription further comprise a buffer or a metal metal salt (e.g., an M'' or M" salt, such as magnesium salt (e.g., an M' or M" salt, such as magnesium chloride), or chloride). In yet other embodiments, the composition com both. prises the at least one reverse transcription primer, the at least 0057. In some embodiments, the composition comprises one pair of forward and reverse primers, no reverse tran the reverse transcriptase, the at least one reverse transcription Scriptase, and no DNA polymerase, where the composition primer, deoxyribonucleotide triphosphates and optionally a can further comprise deoxyribonucleotide triphosphates and buffer and/or a metal salt (e.g., an M' or M" salt, such as optionally a buffer and/or a metal salt (e.g., an M' or M" magnesium chloride) for performing reverse transcription. In salt, such as magnesium chloride). A kit containing reagents other embodiments, the composition comprises the reverse for performing RT-PCR can contain, for example: a compo transcriptase and no reverse transcription primer, where the sition comprising the reverse transcriptase, the DNA poly composition can further comprise deoxyribonucleotide triph merase, the at least one reverse transcription primer, the at osphates and optionally a buffer and/or a metal salt (e.g., an least one pair of forward and reverse primers, deoxyribo M' or M" salt, such as magnesium chloride). In yet other nucleotide triphosphates, and optionally a buffer and/or a embodiments, the composition comprises the at least one metal salt; or (i) a composition comprising the reverse tran reverse transcription primer and no reverse transcriptase, Scriptase, the DNA polymerase, no reverse transcription where the composition can further comprise deoxyribonucle primer, and no pair of forward and reverse primers, which can otide triphosphates and optionally a buffer and/or a metal salt further comprise deoxyribonucleotide triphosphates and (e.g., an M' or M" salt, such as magnesium chloride). A kit optionally a buffer and/or a metal salt; and (ii) a separate containing reagents for performing reverse transcription can composition comprising the at least one reverse transcription contain, for example: a composition comprising the reverse primer, the at least one pair of forward and reverse primers, no transcriptase, the at least one reverse transcription primer, reverse transcriptase, and no DNA polymerase, which can deoxyribonucleotide triphosphates and optionally a buffer further comprise deoxyribonucleotide triphosphates and and/or a metal salt; or (i) a composition comprising the optionally a buffer and/or a metal salt. reverse transcriptase and no reverse transcription primer, 0061. In some embodiments, the composition comprises which can further comprise deoxyribonucleotide triphos one or more RT-PCR reagents for amplifying at least one phates and optionally a buffer and/or a metal salt; and (ii) a polydeoxyribonucleotide complementary to at least one separate composition comprising the at least one reverse tran nucleic acid (e.g., genetic) sequence of the RNA of an RNA Scription primer and no reverse transcriptase, which can fur virus, such as the RNA of any RNA virus described herein. A ther comprise deoxyribonucleotide triphosphates and option kit can contain a composition comprising all of the RT-PCR ally a buffer and/or a metal salt. reagents for amplifying at least one poly deoxyribonucleotide 0058. In further embodiments, the composition may com complementary to at least one nucleic acid (e.g., genetic) prise one or more reagents for performing reverse transcrip sequence of the RNA of an RNA virus, or two or more tion PCR (RT-PCR), wherein the reagents for performing compositions that in total comprise all of the RT-PCR RT-PCR comprise a reverse transcriptase, a DNA poly reagents. merase, at least one reverse transcription primer for reverse 0062. In additional embodiments, the composition com transcribing at least one polyribonucleotide to produce at prises reagents for performing transcription, wherein the least one poly deoxyribonucleotide complementary to the at reagents for performing transcription comprise an RNA poly least one polyribonucleotide, and at least one pair of a for merase. In further embodiments, the reagents for performing ward primer and a reverse primer for amplifying the at least transcription further comprise ribonucleotide triphosphates. one complementary polydeoxyribonucleotide; and the at In certain embodiments, the reagents for performing tran least one reverse transcription primer optionally is labeled scription further comprise a buffer or a metal salt (e.g., an M' with a dye (e.g., a fluorescent dye) and the at least one pair of or M" salt, such as magnesium chloride), or both. forward primer and reverse primer optionally is labeled with 0063. In further embodiments, the composition comprises a dye (e.g., a fluorescent dye). a polypeptide that regulates (e.g., agonizes or antagonizes/ 0059. In certain embodiments, the DNA polymerase com inhibits) an enzyme. Non-limiting examples of polypeptides prises a DNA polymerase that is stable at elevated tempera that regulate enzymes include polypeptides that inhibit pro ture (e.g., at about 60° C., 70° C., 80° C., 90° C. or higher), teases, such as the protease inhibitors described herein. Such as a Taq polymerase. In additional embodiments, the 0064. In other embodiments, the composition comprises reagents for performing RT-PCR further comprise deoxyri an antibody. In some embodiments, the antibody is used in an bonucleotide triphosphates. In certain embodiments, the immunoassay. In certain embodiments, the immunoassay is reagents for performing RT-PCR further comprise a buffer or an enzyme-linked immunosorbent assay (ELISA) or a sand a metal salt (e.g., an M'' or M" salt, such as magnesium wich immunoassay. In further embodiments, the antibody chloride), or both. used in an immunoassay comprises an unlinked antibody, an 0060. In some embodiments, the composition comprises antibody bound to a solid Substrate (e.g., a bead), an antibody the reverse transcriptase, the DNA polymerase, the at least conjugated to a detection protein or enzyme or a fragment one reverse transcription primer, the at least one pair of for thereof, or any combination thereof, wherein the antibody ward and reverse primers, deoxyribonucleotide triphos may or may not be labeled with a dye (e.g., a fluorescent dye, phates, and optionally a buffer and/or a metal salt (e.g., an a chemiluminescent dye, or a phosphorescent dye). M' or M" salt, such as magnesium chloride) for performing 0065. In some embodiments, the composition comprises RT-PCR. In other embodiments, the composition comprises one or more reagents for performing an immunoassay, the reverse transcriptase, the DNA polymerase, no reverse wherein the one or more reagents for performing an immu transcription primer, and no pair of forward and reverse prim noassay comprise an antibody that has affinity for or is spe ers, where the composition can further comprise deoxyribo cific for a target antigen or analyte, the antibody is labeled nucleotide triphosphates and optionally a buffer and/or a with a dye (e.g., a fluorescent dye, a chemiluminescent dye, or US 2015/0267245 A1 Sep. 24, 2015 a phosphorescent dye) or is conjugated to a detection protein limiting examples of proteins and enzymes useful for detec or enzyme or a fragment thereof, and the antibody optionally tion include: phycobiliproteins, phycoerythrins, B-phyco is bound to a solid Substrate (e.g., a bead). erythrin, R-phycoerythrin, and conjugates thereof (e.g., 0066. In further embodiments, the composition comprises phycoerythrin-Streptavidin conjugates, phycoerythrin-alka one or more reagents for performing a sandwich immunoas line phosphatase conjugates, and phycoerythrin-antibody say, wherein: the reagents for performing a sandwich immu conjugates); Streptavidin, avidin, deglycosylated avidin, and noassay comprise a first antibody that has affinity for or is conjugates thereof (e.g., streptavidin-horse radish peroxidase specific for a target antigen or analyte and a second antibody conjugates and Streptavidin-antibody conjugates); peroxi that has affinity for or is specific for the target antigen or dases, horseradish peroxidase, and conjugates thereof (e.g., analyte; the second antibody is labeled with a dye (e.g., a horseradish peroxidase-antibody conjugates); and phos fluorescent dye, a chemiluminescent dye, or a phosphores phatases, alkaline phosphatase, and conjugates thereof (e.g., cent dye) or is conjugated to a detection protein or enzyme or alkaline phosphatase-antibody conjugates). a fragment thereof; and the first antibody optionally is bound 0072. In additional embodiments, the composition com to a solid Substrate (e.g., a bead). prises a polypeptide aptamer that binds specifically to a target 0067. In some embodiments, the composition comprises ligand (e.g., a small molecule, a protein, a nucleic acid, a cell, the first antibody and the second antibody. In other embodi a tissue oran organism). In some embodiments, the polypep ments, the composition comprises the first antibody and not tide aptamer comprises a variable peptide domain or loop the second antibody. In yet other embodiments, the compo sition comprises the second antibody and not the first anti (e.g., a domain or loop containing about 10 to about 20 amino body. A kit containing reagents for performing a sandwich acids) attached at both ends to a polypeptide scaffold (e.g., a immunoassay can contain, for example: a composition com protein scaffold, Such as thioredoxin A). prising the first antibody and the second antibody; or (i) a 0073. The composition can also contain other kinds of composition comprising the first antibody and not the second polypeptides, including without limitation receptors (e.g., antibody; and (ii) a separate composition comprising the sec peripheral membrane proteins, transmembrane proteins and ond antibody and not the first antibody. nuclear receptors), polypeptide ligands (e.g., polypeptide 0068. In additional embodiments, the composition com ligands of antibodies), regulatory factors, hormones, cytok prises one or more reagents for performing an enzyme-linked ines (e.g., interferons and interleukins), structural proteins immunosorbent assay (ELISA), wherein the reagents for per (e.g., collagen and elastin), and toxins. Non-limiting forming ELISA comprise: a detection antibody that has affin examples of hormones include adrenocorticotropic hormone, ity for or is specific for a target antigen or analyte and is angiotensin II, antidiuretic hormone (vasopressin), basic conjugated to a detection enzyme or a fragment thereof, or a fibroblast growth factor-2, cholecystokinin, colony-stimulat first antibody that has affinity for or is specific for a target ing factors (e.g., granulocyte colony-stimulating factor), gas antigen or analyte, and a second antibody that has affinity for trin, growth hormone, insulin, leptin, atrial natriuretic pep or is specific for the first antibody and is conjugated to a tide, brain natriuretic peptide, C-type natriuretic peptide, detection enzyme or a fragment thereof. oxytocin, parathyroid hormone-related protein, prolactin, 0069. In some embodiments, the composition comprises and somatostatin. Examples of toxins include without limi the detection antibody. In other embodiments, the composi tation cyanotoxins, cytotoxins, exotoxins (e.g., botulinum tion comprises the first antibody and the second antibody. In toxin and Corynebacterium diphtheriae exotoxin), hemotox yet other embodiments, the composition comprises the first ins, hepatotoxins (e.g., amatoxins and phallotoxins), myc antibody and not the second antibody. In still other embodi otoxins, necrotoxins, neurotoxins (e.g., bungarotoxins, chlo ments, the composition comprises the second antibody and rotoxin, conotoxins and tetanus toxin), plant toxins (e.g., not the first antibody. A kit containing reagents for perform ricin), insect toxins (e.g., apitoxin), and Snake toxins e.g., ing an ELISA can contain, for example: a composition com cardiotoxins, myotoxins, neurotoxins (such as alpha-neuro prising the detection antibody; or a composition comprising toxins, beta-neurotoxins and dendrotoxins), Sarafotoxins, the first antibody and the second antibody; or (i) a composi hydrolases (such as phosphodiesterases and phospholipases), tion comprising the first antibody and not the second anti lyases, oxydoreductases (such as L-amino acid oxidases), body; and (ii) a separate composition comprising the second transferases, hemorrhagins, hyaluronidases, thrombin-like antibody and not the first antibody. pro-coagulants, and kallikrein-like serine proteases. 0070 The detection protein or enzyme or a fragment 0074. Furthermore, the composition can preserve thereof that is conjugated to an antibody used in an immu polypeptide therapeutics (e.g., hormone therapeutics, cytok noassay can be any protein or enzyme or any fragment thereof ine therapeutics, antibody therapeutics, fusion protein thera that is suitable for detection. In certain embodiments, the peutics, antithrombotics, and toxin therapeutics) and vac detection protein or enzyme or a fragment thereof that is cines in a Substantially water-free fluid medium, e.g., without conjugated to an antibody used in an immunoassay is selected the need for refrigeration. Hormone therapeutics include from the group consisting of phycobiliproteins, phycoeryth without limitation erythropoietin, growth hormone, insulin, rins, B-phycoerythrin, R-phycoerythrin, and fragments and and other hormones described herein. Cytokine therapeutics conjugates thereof. Streptavidin, avidin, deglycosylated avi include without limitation interferons e.g., interferon alpha din, and fragments and conjugates thereof; peroxidases, (including interferon alpha-2a and interferon alpha-2b). horseradish peroxidase, and fragments and conjugates interferon beta (including interferon beta-1a and interferon thereof, and phosphatases, alkaline phosphatase, and frag beta-1b), and derivatives thereof (including interferons ments and conjugates thereof. derivatived with polyethylene glycol (PEG)) and interleu 0071. In further embodiments, the composition comprises kins (e.g., interleukin 2 and interleukin 12). Non-limiting a protein or enzyme useful for detection, where the protein or examples of antibody therapeutics include adalimumab, enzyme may or may not be conjugated to an antibody. Non bevacizumab, infliximab, trastuzumab, and ustekinumab. US 2015/0267245 A1 Sep. 24, 2015

Fusion protein therapeutics include without limitation abata sample comprises whole or fractionated animal (e.g., mam cept, alefacept, denileukin diftitoX, and etanercept. malian, Such as human) blood. In further embodiments, the 0075 Non-limiting examples of antithrombotics include biological sample comprises whole or fractionated animal anti-platelet agents (e.g., abciximab), anticoagulants (e.g., (e.g., mammalian, Such as human) plasma. In still further antithrombin, batroXobin, hementin, hirudin, lepirudin and embodiments, the biological sample comprises whole or frac bivalirudin), and thrombolytics e.g., tissue plasminogen tionated animal (e.g., mammalian, Such as human) serum. activators (including alteplase, reteplase and tenecteplase), I0081. In additional embodiments, the biological sample anistreplase, Streptokinase and urokinase. Examples of toxin comprises cells. A cell can be, e.g., a eukaryotic or prokary therapeutics include without limitation botulinum toxin, otic cell from any single-celled or multi-celled organism, and chlorotoxin, and toxoids used as vaccines (e.g., against botu can be of any type. In some embodiments, the biological lism, diphtheria and tetanus). Non-limiting examples of vac sample comprises animal cells, mammalian cells, human cines include vaccines against botulism, bubonic plague, cells, plant cells, microbial cells, pathogenic cells, bacterial chicken pox, cholera, diphtheria, hepatitis, influenza, cells, fungal cells, protozoan cells or viral particles, or any measles, mumps, polio, rabies, rubella, Small pox, tetanus, combination thereof, or lysates or extracts thereof. The cells tuberculosis, typhoid, and yellow fever. or viral particles can be dissolved or Suspended in a natural 0076. In some embodiments, the composition contains a fluid or a laboratory culture medium (e.g., Dulbecco's phos pharmaceutical formulation comprising a polypeptide and phate buffered saline with 2% fetal bovine serum, Eagle's optionally one or more other Substances as described herein. minimum essential medium (EMEM), Dulbecco's modified In certain embodiments, the concentration of the polypeptide Eagle's medium (DMEM), or the allantoic fluid of embryo in the composition is at least about 10 mg/mL, 50 mg/mL, 100 nated chicken eggs) and then transferred to the Substantially mg/mL, 200 mg/mL,300 mg/mL, 400 mg/mL, 500 mg/mL or water-free fluid medium containing the at least one alcohol 1 g/mL, or about 100-1000 mg/mL, 100-500 mg/mL or 500 Solvent (or the at least one ionic organic solvent in other 1000 mg/mL. In some embodiments, the concentration of the embodiments of a composition comprising a biological mate polypeptide in the composition is at least about 100 mg/mL, rial in a substantially water-free fluid medium, as described or about 100-1000 mg/mL. below). 0077. In further embodiments, the composition comprises I0082 In some embodiments, the biological sample com a catalytic polynucleotide. In some embodiments, the cata prises a bacterium. The bacterium can be non-pathogenic or lytic polynucleotide is a natural or synthetic ribozyme (or pathogenic. Non-limiting examples of bacteria include Bacil RNA enzyme or catalytic RNA). Non-limiting examples of lus (e.g., Bacillus anthracis, Bacillus cereus and Bacillus natural ribozymes include peptidyl transferase 23S rRNA, thuringiensis); Bordetella (e.g., Bordetella pertussis); Borre RNase P. Group I introns, Group II introns, GIR1 branching lia (e.g., Borrelia burgdorferi); Brucella (e.g., Brucella abor ribozyme, leadzyme, hairpin ribozyme, hammerhead tus, Brucella Canis, Brucella melitensis, and Brucella suis); ribozyme, HDV ribozyme, mammalian CPEB3 ribozyme, Campylobacter (e.g., Campylobacter jejuni); Chlamydia and VS ribozyme, glimS ribozyme, and CoTC ribozyme. Chlamydophila (e.g., Chlamydia pneumoniae, Chlamydia Examples of synthetic ribozymes include without limitation trachomatis, and Chlamydophila psittaci); Clostridium (e.g., ribozymes produced from RNA polymerases (e.g., Round 18 Clostridium botulinum, Clostridium difficile, Clostridium RNA polymerase ribozyme and variants thereof, such as perfingens, and Clostridium tetani); Corynebacterium (e.g., B6.61 ribozyme and tC19Z ribozyme) and ribozymes pro Corynebacterium diphtheriae): Enterococcus (e.g., Entero duced from RNA ligases. In other embodiments, the catalytic coccus faecalis and Enterococcus faecium); Escherichia polynucleotide is a deoxyribozyme (or DNA enzyme or cata (e.g., Escherichia coli); Francisella (e.g., Francisella tulla lytic DNA), including deoxyribozymes that catalyze DNA rensis); Haemophilus (e.g., Haemophilus influenzae); Heli phosphorylation, DNA adenylation, DNA deglycosylation, cobacter (e.g., Helicobacter pylori); Legionella (e.g., DNA cleavage, thymine dimer photoreversion, and porphyrin Legionella pneumophila); Leptospira (e.g., Leptospira inter metalation. rogans); Listeria (e.g., Listeria monocytogenes); Mycobacte 0078. In additional embodiments, the composition com rium (e.g., Mycobacterium leprae, Mycobacterium tubercu prises a polynucleotide that binds specifically to a target losis, and Mycobacterium ulcerans); Mycoplasma (e.g., ligand (e.g., a small molecule, a protein, a nucleic acid, a cell, Mycoplasma pneumoniae); Neisseria (e.g., Neisseria gonor a tissue or an organism). In some embodiments, the poly rhoeae and Neisseria meningitidis); Pseudomonas (e.g., nucleotide that binds specifically to a target ligand is a natural Pseudomonas aeruginosa); Rickettsia (e.g., Rickettsia rick or synthetic nucleic acid aptamer (e.g., DNA aptamer, RNA ettsii); Salmonella (e.g., Salmonella typhi and Salmonella aptamer or XNA aptamer). typhimurium); Shigella (e.g., Shigella sonnei); Staphylococ 0079 Non-limiting examples of nucleic acid aptamers cus (e.g., Staphylococcus aureus, Staphylococcus epidermi include DNA aptamers and RNA aptamers that bind dopam dis, and Staphylococcus saprophyticus); Streptococcus (e.g., ine, hemin, HIV trans-acting responsive element, interferons Streptococcus agalactiae, Streptococcus pneumoniae, and (e.g., interferon-gamma), lysozymes, mycotoxins, thrombin, Streptococcus pyogenes); Treponema (e.g., Treponema palli and vascular endothelial growth factor (VEGF). dum); Vibrio (e.g., Vibrio cholerae and Vibrio parahaemolyti 0080. In other embodiments, the composition comprises a cus); and Yersinia (e.g., Yersinia pestis). biological sample. The biological sample can be, e.g., a clini I0083. In other embodiments, the biological sample com cal sample, a Surgical sample, a laboratory sample, a research prises a fungus. The fungus can be non-pathogenic or patho sample, a forensic sample, a veterinary sample, an environ genic. Examples of fungi include without limitation Aspergil mental sample, an agricultural sample, or an industrial lus (e.g., Aspergillus Clavatus, Aspergillus fumigatus, and sample. The biological sample can be re-hydrated by addition Aspergillus flavus); Blastomyces (e.g., Blastomyces dermati of water or an aqueous solution (e.g., an aqueous buffer) for tidis); Candida (e.g., Candida albicans); Coccidioides (e.g., analysis, if desired. In some embodiments, the biological Coccidioides immitis and Coccidioides posadasii); Crypto US 2015/0267245 A1 Sep. 24, 2015

COccus (e.g., Cryptococcus albidus, Cryptococcus gattii, agent and hepatitis G. virus), and Pestivirus (e.g., bovine viral Cryptococcus laurentii, and Cryptococcus neoformans); diarrhea virus, classical Swine fever virus, and hog cholera Fusarium (e.g., Fusarium graminearum, Fusarium virus); orthomyxoviruses including Influenzavirus A (e.g., Oxysporum, Fusarium proliferatum, Fusarium Solani com influenza A virus), Influenzavirus B (e.g., influenza B virus), plex, and Fusarium verticillioides); Histoplasma (e.g., His Influenzavirus C (e.g., influenza C virus), Isavirus (e.g., toplasma capsulatum); Pneumocystis e.g., Pneumocystis infectious salmon anemia virus), and Thogotovirus (e.g., jirovecii (or Pneumocystis Carinii); Stachybotrys (e.g., Dhori virus and Thogoto virus); paramyxoviruses including Stachybotry's chartarum); TrichospOron (e.g., TrichospOron Aquaparamyxovirus (e.g., Atlantic salmon paramyxovirus asahi, TrichospOron asteroides, TrichospOron cutaneum, and Pacific salmon paramyxovirus), AVulavirus (e.g., New TrichospOron dermatis, TrichospOron dohaense, Trichos castle disease virus), Ferlavirus (e.g., Fer-de-Lance virus), poron inkin, TrichospOron loubieri, TrichospOron mucoides Henipavirus (e.g., hendravirus and nipahvirus), Morbillivirus and TrichospOron ovoides); and Zygomycetes (e.g., Rhizopus (e.g., measles virus, canine distemper virus, Ovine rinderpest stolonifer). virus, phocine distemper virus, and rinderpest virus), 0084. In yet other embodiments, the biological sample Respirovirus (e.g., human parainfluenza viruses 1 and 3, and comprises a protozoan. The protozoan can be non-pathogenic Sendai virus), Rubulavirus (e.g., mumps virus, human parain or pathogenic. Examples of protozoa include without limita fluenza viruses 2 and 4, Menangle virus, simian parainfluenza tion Balantidium (e.g., Balantidium coli); Cryptosporidium virus 5, Tioman virus, and Tuhokoviruses 1, 2 and 3), TPMV (e.g., Cryptosporidium canis, Cryptosporidium fells, like viruses (e.g., Mossman virus, Nariva virus, Salem virus, Cryptosporidium hominis, Cryptosporidium meleagridis, and Tupaia paramyxovirus), Beilong virus, Pneumovirus Cryptosporidium muris and Cryptosporidium parvum); (e.g., human respiratory syncytial virus and bovine respira Entamoeba (e.g., Entamoeba dispar and Entamoeba his tory syncytial virus), Metapneumovirus (e.g., human metap tolytica); Giardia (e.g., Giardia lamblia and Giardia muris); neumovirus and avian pneumovirus), J virus, Sunshine virus, Leishmania (e.g., Leishmania braziliensis, Leishmania and Tailam virus; picornaviruses including Aphthovirus infantum and Leishmania major); Naegleria (e.g., Naegleria (e.g., foot-and-mouth disease virus, bovine rhinitis A virus, fowleri); Plasmodia (e.g., Plasmodia falciparum, Plasmodia bovine rhinitis B virus, and equine rhinitis A virus), Avihepa knowlesi, Plasmodia malariae and Plasmodia vivax); Toxo tovirus (e.g., duck hepatitis. A virus), Cardiovirus (e.g., plasma (e.g., Toxoplasma gondii); Trichomonas (e.g., Tri encephalomyocarditis virus and Theilovirus), Enterovirus chomonas vaginalis); and Trypanosoma (e.g., Trypanosoma (e.g., human enteroviruses A to D, simian enterovirus A, bruceii and Trypanosoma Cruzi). bovine enterovirus, porcine enterovirus B, and human rhi 0085. In additional embodiments, the biological sample noviruses A to C), Erbovirus (e.g., equine rhinitis B virus), comprises a virus. The virus can be a DNA virus or an RNA Hepatovirus (e.g., hepatitis. A virus), Kobuvirus (e.g., Aichi virus. Non-limiting examples of DNA viruses include aden virus and bovine kobuvirus), Parechovirus (e.g., human pare oviruses including Atadenovirus (e.g., Ovine adenovirus D), chovirus and Ljungan virus), Salivirus (e.g., Salivirus A). Aviadenovirus (e.g., fowl adenovirus A), Ichtadenovirus Sapelovirus (e.g., avian Sapelovirus, porcine Sapelovirus, and (e.g., Sturgeon adenovirus A), Mastadenovirus (e.g., human simian Sapelovirus), Senecavirus (e.g., Seneca Valley virus), adenovirus C and AD-36), and Siadenovirus (e.g., frog aden Teschovirus (e.g., porcine teschovirus), and Tremovirus (e.g., ovirus); hepadnaviruses including Orthohepadnavirus (e.g., avian encephalomyelitis virus); retroviruses including hepatitis B virus) and Avihepadnavirus (e.g., duck hepatitis B Alpharetrovirus (e.g., avian leukosis virus and rous sarcoma virus); herpesviruses including human herpesviruses (e.g., virus), Betaretrovirus (e.g., mouse mammary tumour virus), herpes simplex virus-1, herpes simplex virus-2. Varicella Gammaretrovirus (e.g., murine leukemia virus and feline leu Zoster virus, Epstein-Barr virus, cytomegalovirus, roseolovi kemia virus), Deltaretrovirus (e.g., human T-lymphotropic rus, herpes lymphotropic virus, pityriasis rosea virus, and virus and bovine leukemia virus), Epsilon retrovirus (e.g., Kaposi's sarcoma-associated herpesvirus) and Zoonotic her Walleye dermal sarcoma virus), Lentivirus (e.g., human pesviruses (e.g., cercopithecine herpesvirus-1 and murine immunodeficiency viruses (HIV), simian immunodeficiency gammaherpesvirus-68); papillomaviruses (e.g., human pap viruses, and feline immunodeficiency viruses), and Spuma illomaviruses 1 to 18); and polyomaviruses including Ortho virus (e.g., simian foamy virus); rhabdoviruses including polyomavirus (e.g., simian virus 40, B-lymphotropic polyo Cytorhabdovirus (e.g., lettuce necrotic yellows virus), mavirus, baboon polyomavirus 1, bat polyomavirus, BK Dichorhabdovirus (e.g., orchid fleck virus), Ephemerovirus polyomavirus, Bornean orang-utan polyomavirus, Sumatran (e.g., bovine ephemeral fever virus), Lyssavirus (e.g., rabies orang-utan polyomavirus, bovine polyomavirus, California virus), Novirhabdovirus (e.g., infectious hematopoietic sea lion polyomavirus, chimpanzee polyomavirus, hamster necrosis virus), Nucleorhabdovirus (e.g., potato yellow dwarf polyomavirus, JC polyomavirus, Merkel cell polyomavirus, virus), and Vesiculovirus (e.g., vesicular stomatitis Indiana murine pneumotropic virus, murine polyomavirus, squirrel virus); and togaviruses including Rubivirus (e.g., rubella monkey polyomavirus, and trichodysplasia spinuolsa-associ virus) and Alphavirus (e.g., Chikungunya virus, Eastern ated polyomavirus), Wukipolyomavirus (e.g., human polyo equine encephalitis virus, Western equine encephalitis virus, maviruses 6 and 7, KI polyomavirus, and WU polyomavirus), Venezuelan equine encephalitis virus, Onyongnyong virus, Avipolyomavirus (e.g., avian polyomavirus, canary polyo Ross River virus, Semliki Forest virus, and Sindbis virus). mavirus, crow polyomavirus, finch polyomavirus, and goose I0087. In some embodiments, the composition containing hemorrhagic polyomavirus), and human polyomavirus 9. the biological material further comprises one or more Sub I0086 Non-limiting examples of RNA viruses include stances selected from the group consisting of reducing agents, coronaviruses including human coronaviruses (e.g., SARS antioxidants, free radical scavengers, oxygen radical scaven coronavirus); flaviviruses including Flavivirus (e.g., yellow gers, hydroxyl radical scavengers, singlet oxygen quenchers, fever virus, West Nile virus, and dengue fever virus), Hepa hydroperoxide-removing agents, protease inhibitors, civirus (e.g., hepatitis C virus), Hepatitis GVirus (e.g., the GB nuclease inhibitors, ribonuclease (RNase) inhibitors, deox US 2015/0267245 A1 Sep. 24, 2015 yribonuclease (DNase) inhibitors, metal chelators, preserva 0092. Examples of hydroperoxide-removing agents tives, anti-microbials, buffers (or buffering agents), deter include without limitation catalase, glutathione, peroxidases, gents, and chaotropes. The composition can comprise the one glutathione peroxidases, pyruvate, and analogs, derivatives or more Substances in appropriate amounts to enhance, e.g., and salts thereof. the stability and/or the solubility of the biological material in 0093. Non-limiting examples of protease inhibitors the substantially water-free fluid medium. It is understood include aspartic protease inhibitors, cysteine protease inhibi that a Substance can have one or more functions or properties. tors, metalloprotease inhibitors, serine protease inhibitors, As an example, a Substance can be a reducing agent, an threonine protease inhibitors, trypsin inhibitors (e.g., avian antioxidant, a free radical scavenger, an oxygen radical scav egg white trypsin inhibitors, bovine trypsin inhibitors, lima enger, a hydroxyl radical scavenger or a singlet oxygen bean trypsin inhibitors, and soybean trypsin inhibitors such as quencher, or any combination thereof. As another example, a Kunitz trypsin inhibitor and Bowman-Birk inhibitor), substance can be a metal chelator, a DNase inhibitor or an Kunitz-type protease inhibitors, 4-(2-aminoethyl)benzene anti-microbial, or any combination thereof. sulfonyl fluoride (AEBSF) and salts thereof (e.g., HCl salt), 0088. Examples of reducing agents, antioxidants, and free amastatin, antithrombin III, antipain, APMSF, aprotinin, radical scavengers include without limitation cysteine, bestatin, benzamidine, calpain inhibitors I and II, chymosta dithionite, dithioerythritol, dithiothreitol (DTT), dysteine, tin, 3,4-dichloroisocoumarin, diisopropyl fluorophosphate 2-mercaptoethanol, mercaptoethylene, bisulfite, sodium met (DFP), E-64, elastatinal, hirustasin, leupeptin, alpha-2-mac abisulfite, pyrosulfite, pentaerythritol, thioglycolic acid, urea, roglobulin, Pefabloc SC, pepstatin, 1,10-phenanthroline, uric acid, vitamin C. vitamin E. Superoxide dismutases, and phosphoramidon, phenylmethylsulfonyl fluoride (PMSF), analogs, derivatives and salts thereof. PMSF Plus, tissue inhibitors of metalloproteinases (e.g., 0089. Non-limiting examples of oxygen radical scaven TIMP-1, TIMP-2, TIMP-3 and TIMP-4), tosyllysine chlo gers include Sugar alcohols (e.g., erythritol, mannitol, Sorbi romethyl ketone (TLCK) and salts thereof (e.g., HCl salt), tol, and Xylitol), monosaccharides (e.g., hexoses, allose, tosyl phenylalanyl chloromethyl ketone (TPCK), and ana altrose, fructose, fucose, fuculose, galactose, glucose, gulose, logs, derivatives and salts thereof. idose, mannose, rhamnose, Sorbose, tagatose, talose, pen 0094. Non-limiting examples of RNase inhibitors include toses, arabinose, lyxose, ribose, deoxyribose, ribulose, mammalian ribonuclease inhibitor proteins e.g., porcine Xylose, Xylulose, tetroses, erythrose, erythrulose, and ribonuclease inhibitor and human ribonuclease inhibitor threose), disaccharides (e.g., cellobiose, lactose, maltose, (e.g., human placentaribonuclease inhibitor and recombinant Sucrose, and trehalose), complex Sugars (e.g., trisaccharides, human ribonuclease inhibitor), aurintricarboxylic acid kestose, isomaltotriose, maltotriose, maltotriulose, melezi (ATA) and salts thereof e.g., triammonium aurintricarboxy tose, nigerotriose, raffinose, tetrasaccharides, stachyose, late (aluminon), adenosine 5'-pyrophosphate, 2'-cytidine fructo-polysaccharides, galacto-polysaccharides, mannan monophosphate free acid (2'-CMP), 5'-diphosphoadenosine polysaccharides, gluco-polysaccharides, glycogen, starch, 3'-phosphate (pp.A-3'-p), 5'-diphosphoadenosine 2'-phos amylose, amylopectin, dextrin, cellulose, glucans, beta-glu phate (pp.A-2'-p), leucine, oligovinysulfonic acid, poly(as cans, dextran, fructans, inulin, glucosamine polysaccharides, partic acid), tyrosine-glutamic acid polymer, 5'-phospho-2'- chitin, aminoglycosides, apramycin, gentamycin, kanamy deoxyuridine 3'-pyrophosphate P->5'-ester with adenosine cin, netilmicin, neomycin, paromomycin, Streptomycin, 3'-phosphate (pdUppAp), and analogs, derivatives and salts tobramycin, glycosaminoglycans (mucopolysaccharides), thereof. chondroitin Sulfate, dermatan Sulfate, keratan Sulfate, hep 0.095 Examples of DNase inhibitors and metal chelators arin, heparan Sulfate, and hyaluronan), and analogs, deriva include without limitation aurintricarboxylic acid (ATA) and tives and salts thereof. salts thereof e.g., triammonium aurintricarboxylate (alumi 0090. Examples of hydroxyl radical scavengers include non), boric acid, borate, citric acid, citrate, Salicylic acid, without limitation azides (e.g., sodium azide), cysteine, dim salicylate, 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tet ethylsulfoxide, histidine, Salicylic acid, Salicylate, Sugar alco raacetic acid (BAPTA), diethylene triamine pentaacetic acid hols (e.g., erythritol, mannitol, Sorbitol, and Xylitol), (DTPA), ethylenediaminetetraacetic acid (EDTA), ethylene monosaccharides (e.g., those described herein), disaccha glycol tetraacetic acid (EGTA), glycoletherdiaminetetraace rides (e.g., cellobiose, lactose, maltose, Sucrose, and treha tic acid (GEDTA), N-(2-hydroxyethyl)ethylenediamine-N, lose), complex Sugars (e.g., those described herein), and ana N',N'-triacetic acid (HEDTA), nitrilotriacetic acid (NTA), logs, derivatives and salts thereof. 2,2'-bipyridine, o-phenanthroline, triethanolamine, and ana 0091. Non-limiting examples of singlet oxygen quenchers logs, derivatives and salts thereof. include azides (e.g., Sodium azide), ascorbic acid, ascorbate, 0096. Examples of preservatives include without limita alkyl imidazoles (e.g., carnosine, histamine, histidine, and tion azides (e.g., Sodium azide), polyethylene glycol (PEG), imidazole 4-acetic acid), indoles (e.g., tryptophan and deriva and anti-microbials (e.g., anti-biotic, anti-fungal, anti-para tives thereof, such as N-acetyl-5-methoxytryptamine, sitic and anti-viral agents). Anti-microbials include without N-acetyl serotonin, and 6-methoxy-1,2,3,4-tetrahydro-beta limitation beta-lactams, penicillins, semi-synthetic penicil carboline), Sulfur-containing amino acids (e.g., cysteine, lins, mono-bactams, carboxypenems, aminoglycosides, gly S-allyl-cysteine, S-aminoethyl-cysteine, djenkolic acid, copeptides, lincomycins, macrollides, allylamines, azoles, ethionine, methionine, N-formyl-methionine, lanthionine, polyenes, tetraenes, Sulfonamides, pyrimidines, thiocarbam and felinine), phenolic compounds (e.g., tyrosine and deriva ates, benzoic acid compounds, rifamycins, tetracyclines, tives thereof), aromatic carboxylic acids (e.g., salicylic acid reverse transcriptase inhibitors, protease inhibitors, thymi and derivatives thereof), vitamin A and derivatives thereof dine kinase inhibitors, glycoprotein synthesis inhibitors, (e.g., carotenoids, beta-carotene, retinol and retinal), Vitamin Sugar synthesis inhibitors, glucan synthesis inhibitors, struc E and derivatives thereof (e.g., tocopherols, alpha-tocopherol tural protein synthesis inhibitors, viral maturation inhibitors, and tocotrienols), and analogs, derivatives and salts thereof. nucleoside analogs, polypeptides, and analogs, derivatives US 2015/0267245 A1 Sep. 24, 2015

and salts thereof. For example, anti-microbials include with chain alcohols, modified oxypropylated Straight-chain alco out limitation penicillin, cephalosporin, amplicillin, amoxy hols, polyethylene glycol mono-oleate compounds, cillin, aztreonam, clavulanic acid, imipenem, streptomycin, polysorbate compounds (e.g., polyoxyethylene Sorbitan gentamycin, Vancomycin, clindamycin, polymyxin, erythro monolaurate compounds, such as Polysorbates (Tweens) 20, mycin, bacitracin, amphotericin, nystatin, rifampicin, tetra 40, 60 and 80), phenolic fatty alcohol ethers, phenolic poly cycline, chlortetracycline, doxycycline, chloramphenicol, ethylene glycols (e.g., TritonX-100), and analogs, derivatives ammolfine, butenafine, naftifine, terbinafine, ketoconazole, and salts thereof. fluconazole, elubiol, econazole, econaxole, itraconazole, iso 0099 Examples of chaotropes include without limitation conazole, imidazole, miconazole, Sulconazole, clotrimazole, formamide, guanidine and salts thereof (e.g., guanidinium enilconazole, oxiconazole, tioconazole, terconazole, buto hydrochloride), isothiocyanate, urea, and analogs, derivatives conazole, thiabendazole, Voriconazole, Saperconazole, Serta and salts thereof. conazole, fenticonazole, posaconazole, bifonazole, flutrima Zole, nystatin, pimaricin, amphotericin B, flucytosine, 0100. In some embodiments, the composition containing natamycin, tolnaftate, mafenide, dapsone, caspofungin, acto the biological material comprises an oxygen radical scaven funicone, griseofulvin, potassium iodide, Gentian Violet, ger. In certain embodiments, the concentration of the oxygen ciclopiroX, ciclopiroX olamine, haloprogin, silver Sulfadiaz radical scavenger by mass (mg) relative to the Volume (LL) of ine, undecylenate, undecylenic acid, undecylenic alkanola the at least one alcohol solvent (or the at least one ionic mide, Carbol-Fuchsin, nevirapine, delavirdine, efavirenz, organic solvent in other embodiments of a composition com saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, prising a biological material in a Substantially water-free fluid zidovudine (AZT), stavudine (d4T), lamivudine (3TC), medium, as described below) is at least about 1%. 5%, 10%, didanosine (DDI), zalcitabine (ddC), abacavir, acyclovir, 25%, 50%, 75%, 100%, 125%, 150%, 200%, 250% or 300%. penciclovir, Valacyclovir, ganciclovir, and analogs, deriva In some embodiments, the concentration of the oxygen radi tives and salts thereof. cal scavenger by mass (mg) relative to the Volume (LL) of the at least one alcohol solvent (or the at least one ionic organic 0097. In some embodiments, the buffers or buffering solvent in other embodiments) is about 25%-150%, 25%- agents provide buffering in a basic pH range (e.g., about pH 7 125%, 25%400%, 25%-7.5% or 25%-50%. or 8 to 11, about pH 7 or 8 to 10, about pH 7 or 8 to 9, about pH 10-11, about pH 9-10, about pH 8-9, or about pH 7-8). 0101. In some embodiments, the composition comprises Non-limiting examples of buffers or buffering agents that the biological material and an oxygen radical scavenger in provide buffering in a basic pH range include borate, saline ethylene glycol. 1,3-propanediol, glycerol or 1.2-butanediol. phosphate, saline Sodium citrate, 2-(methylamino)Succinic or any combination thereof. In certain embodiments, the oxy acid, N,N-bis(2-hydroxyethyl)glycine (bicine), N-tris(hy gen radical scavenger is mannitol, mannose, Sucrose or tre droxymethyl)methylglycine (tricine), tris(hydroxymethyl) halose. methylamine (Tris), 2-(cyclohexylamino)ethanesulfonic 0102) Infurther embodiments, the composition containing acid (CHES), 4-(2-hydroxyethyl)-1-piperazineethane the biological material comprises one or more Substances that sulfonic acid (HEPES), piperazine-N,N'-bis(2-ethane enhance the stability of single-stranded and double-stranded sulfonic acid) (PIPES), 2-tris(hydroxymethyl)methyl polynucleotides containing RNA nucleotides and/or DNA aminoethanesulfonic acid (TES), 3-amino-1- nucleotides. In certain embodiments, the composition com propanesulfonic acid, 3-(cyclohexylamino)-1- prises: (a) a metal chelator, a hydroxyl radical scavenger, and propanesulfonic acid (CAPS), 3-(cyclohexylamino)-2- an RNase inhibitor; or (b) a hydroxyl radical scavenger and a hydroxy-1-propanesulfonic acid (CAPSO), N-(2- DNase inhibitor. hydroxyethyl)piperazine-N'-(3-propanesulfonic acid) 0103) In additional embodiments, the composition con (EPPS), 3-(N-morpholino)propanesulfonic acid (MOPS), taining the biological material comprises a metal salt, option 3-tris(hydroxymethyl)methylamino-propanesulfonic ally in addition to one or more other substances described acid (TAPS), 3-N-tris(hydroxymethyl)methylamino-2-hy herein. The metal salt can enhance the stability and/or the droxypropanesulfonic acid (TAPSO), 4-(cyclohexylamino)- solubility of the biological material in the substantially water 1-butanesulfonic acid (CABS), and analogs, derivatives and free fluid medium. As an example, the metal salt can increase salts thereof. the melting temperature of a double-stranded polynucleotide 0098. The detergents can be denaturing detergents or non containing RNA nucleotides and/or DNA nucleotides. As denaturing detergents. Non-limiting examples of denaturing another example, the metal salt can increase the solubility and detergents and non-denaturing detergents include anionic the refolding yield, and can promote retention of the activity, Surfactants, cationic Surfactants, non-ionic Surfactants, Zwit of a protein (e.g., an enzyme) preserved in the Substantially terionic Surfactants, ampholytic Surfactants, benzethonium water-free fluid medium. In some embodiments, the metal chloride, cetyltrimethylammonium bromide (CTAB), 3-(3- salt comprises an M'' (or monovalent) salt or an M (or cholamidopropyl)dimethylammonio-1-propanesulfonate divalent) salt, or both. M'' (or monovalent) salts include (CHAPS), 3-(3-cholamidopropyl)dimethylammonio-2-hy without limitation lithium salts, sodium salts and potassium droxy-1-propanesulfonate (CHAPSO), N,N-dimethyl-decy salts of fluoride, chloride, bromide, iodide, acetate, formate, lamine-N-oxide, guanidinium thiocyanate, hexadecyltrim nitrate, perchlorate (CIO), phosphate, sulfate, tetrafluo ethylammonium bromide, lithium dodecyl sulfate (LDS), roborate (BF) and thiocyanate (SCN), and M (or diva sodium dodecyl sulfate (SDS), sodium lauryl sulfate, sodium lent) salts include magnesium salts, manganese salts and cal cholate, sodium deoxycholate, ethoxylated fatty alcohol cium salts of fluoride, chloride, bromide, iodide, acetate, ethers, lauryl ethers, ethoxylated alkyl phenol compounds formate, nitrate, perchlorate, phosphate, Sulfate, tetrafluo e.g., ethoxylated nonyl phenol compounds, such as NP-40 roborate and thiocyanate. In some embodiments, the metal (nonyl phenoxypolyethoxylethanol), octylphenoxy poly salt comprises LiCl, NaCl, KC1, MgCl2 or MnCl, or any ethoxy ethanol compounds, modified oxyethylated Straight combination thereof. US 2015/0267245 A1 Sep. 24, 2015

0104. The biological material is soluble in the substan 3 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 1.5 tially water-free alcohol solvent, and thus may not need to be years or 2 years. In certain embodiments, the biological mate re-dissolved for use in fluid-phase reactions or assays, includ rial retains at least about 90% of its function or activity after ing nucleic acid amplification reactions based on PCR and storage of the composition comprising the biological material analytical and diagnostic assays, Such as immunoassays. In in a closed container (e.g., a capped tube, vial or well) at some embodiments, at least about 50%, 60%, 70%, 80%, ambient temperature for at least about 3 months or 6 months. 90%. 95% or 99% of the biological material by mass is The function or activity of a biological material preserved in dissolved in the Substantially water-free alcohol Solvent, e.g., the substantially water-free alcohol solvent and tested at a after storage of the composition comprising the biological particular time point can be compared to the function or material in a closed container (e.g., a capped tube, vial or activity of a positive control, e.g., the function or activity of well) at a temperature from ambient temperature to about 40° the biological material prepared under appropriate conditions C. for at least about 1 day, 3 days, week, 2 weeks, 3 weeks, 1 (e.g., in an aqueous medium) shortly before its use in the test month, 2 months, 3 months, 6 months, 1 year, 1.5 years or 2 protocol (e.g., test reaction or test assay). years. In certain embodiments, at least about 80% or 90% of 0107 Non-limiting examples of retention of a biological the biological material by mass is dissolved in the Substan material's function or activity include: a polypeptide enzyme tially water-free alcohol solvent after storage of the compo or a polynucleotide enzyme retaining its enzymatic or cata sition comprising the biological material in a closed container lytic function or activity; a polypeptide retaining its ability to (e.g., a capped tube, vial or well) at ambient temperature for regulate (e.g., agonize or antagonize/inhibit) an enzyme; an at least about 3 months or 6 months. antibody, a polypeptide aptamer or a polynucleotide aptamer 0105. Furthermore, the biological material is stable (e.g., retaining its binding affinity or specificity for a target antigen, retains its structural integrity) in the substantially water-free ligand or analyte; a polypeptide ligand of an antibody retain alcohol solvent at ambient temperature or higher, and thus ing its ability to be recognized and bound by the antibody; a does not need to be refrigerated or frozen during shipping or hormone or a cytokine retaining its biological function or storage. In some embodiments, the biological material is activity; a polypeptide therapeutic retaining its pharmaco stable (e.g., retains its structural integrity) in the Substantially logical function or activity; a vaccine retaining its prophylac water-free alcohol solvent after storage of the composition tic or immune function or activity; a pair of forward and comprising the biological material in a closed container (e.g., reverse primers retaining their ability to prime amplification a capped tube, vial or well) at a temperature from ambient of a target poly deoxyribonucleotide or a target nucleic acid temperature to about 40°C. for at least about 1 day, 3 days, 1 (e.g., genetic) locus; a reverse transcription primer retaining week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 its ability to prime reverse transcription of a target polyribo months, 1 year, 1.5 years or years. In further embodiments, nucleotide; a biological sample retaining its biological activ the biological material is resistant to irreversible hydrolytic ity or its function as an analyte in an assay, or components in damage, irreversible oxidative damage, and irreversible dena the biological sample retaining their biological activity or turation (e.g., irreversible unfolding or irreversible loss of their function as analytes in an assay; and bacterial cells secondary structure or tertiary structure) after storage of the retaining their infectivity in an appropriate medium (e.g., an composition comprising the biological material in a closed agar medium or a fluid culture), or viral particles retaining container (e.g., a capped tube, vial or well) at a temperature their infectivity in an appropriate medium (e.g., a natural fluid from ambient temperature to about 40°C. for at least about 1 or a laboratory cell culture). day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 1.5 years or 2 years. In certain Method of Preserving a Biological Material in an Anhydrous, embodiments, the biological material is resistant to irrevers Non-Ionic Organic Solvent ible hydrolytic damage, irreversible oxidative damage, and 0.108 Further embodiments of the disclosure relate to a irreversible denaturation (e.g., irreversible unfolding or irre method of preserving a biological material in a Substantially versible loss of secondary structure or tertiary structure) after water-free, non-ionic organic solvent (e.g., an alcohol Sol storage of the composition comprising the biological material Vent). In some embodiments, the method comprises: mixing in a closed container (e.g., a capped tube, vial or well) at an aqueous mixture comprising a polypeptide, a polynucle ambient temperature for at least about 3 months or 6 months. otide or a biological sample, or any combination thereof, with 0106. In addition, the biological material retains its func at least one alcohol solvent to produce an aqueous organic tion or activity when it is preserved in the substantially water mixture, wherein: the polypeptide comprises an enzyme that free alcohol solvent at ambient temperature or higher and is mediates a nucleic acid reaction, a polypeptide that regulates tested for its function or activity under appropriate conditions an enzyme, an antibody, a polypeptide ligand of an antibody, (e.g., in an aqueous medium). In some embodiments, the a polypeptide aptamer, a protein or enzyme useful for detec biological material retains its function or activity after storage tion, a toxin, a hormone, a cytokine, a polypeptide therapeutic of the composition comprising the biological material in a or a vaccine, or a derivative thereof or any combination closed container (e.g., a capped tube, vial or well) at a tem thereof; the polynucleotide comprises a polynucleotide used perature from ambient temperature to about 40°C. for at least in a nucleic acid reaction, a catalytic polynucleotide, or a about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 polynucleotide that binds specifically to a target ligand, or a months, 3 months, 6 months, 1 year, 1.5 years or 2 years. In derivative thereof or any combination thereof; the biological further embodiments, the biological material retains at least sample comprises a biological fluid, a biological Suspension, about 50%, 60%, 70%, 80%, 90%, 95% or 99% of its function a fluid aspirate, blood, plasma, serum, lymph, cerebrospinal or activity after storage of the composition comprising the fluid, gastric fluid, bile, perspiration, ocular fluid, tears, oral biological material in a closed container (e.g., a capped tube, fluid, sputum, saliva, a buccal sample, a tonsil sample, a nasal vial or well) at a temperature from ambient temperature to sample, mucus, a nasopharyngeal sample, semen, urine, a about 40°C. for at least about 1 day, 3 days, 1 week, 2 weeks, vaginal sample, a cervical sample, a rectal sample, a fecal US 2015/0267245 A1 Sep. 24, 2015 sample, a wound or purulent sample, hair, a tissue, a tissue embodiments, water is removed from the aqueous organic homogenate, cells, a cellular lysate, a tissue or cell biopsy, mixture at reduced pressure and at reduced temperature but skin cells, tumor or cancer cells, a microbe, a pathogen, a above the freezing point of the aqueous organic mixture or the bacterium, a fungus, a protozoan or a virus, or any combina at least one alcohol solvent. In further embodiments, water is tion thereof, and the at least one alcohol solvent is selected removed from the aqueous organic mixture at a relative from the group consisting of linear and branched C2-C6 humidity of no more than about 60%, 50%, 40%, 30% or acyclic alcohols having one or more hydroxyl groups and 20%. C3-C6 cyclic alcohols having one or more hydroxyl groups 0112 The composition can be any composition compris and three to six ring carbon atoms, wherein the acyclic alco ing a biological material in a Substantially water-free alcohol hols and the cyclic alcohols optionally comprise one or more Solvent as described herein. For example, the composition halide atoms; and removing water from the aqueous organic can comprise one or more Substances as described herein, mixture to produce a composition comprising the polypep where the aqueous mixture, the at least one alcohol Solvent or tide, the polynucleotide or the biological sample, or any com the aqueous organic mixture, or any combination thereof, can bination thereof, and the at least one alcohol solvent, wherein comprise the one or more Substances (e.g., the one or more the composition is in a fluid state and is substantially free of Substances can be added to the aqueous mixture, the at least Water. one alcohol solvent or the aqueous organic mixture, or any 0109. In some embodiments, the at least one alcohol sol combination thereof). In certain embodiments, the one or vent in the composition comprises no more than about 20%, more Substances comprise: (a) a protease inhibitor; (b) an 15%, 10%, 5%, 4%, 3%, 2%, 1% or 0.5% water by mass oxygen radical scavenger; (c) a metal chelator, a hydroxyl relative to the combined mass of water and the at least one radical scavenger, and an RNase inhibitor, or (d) a hydroxyl alcohol solvent after removal of water from the aqueous radical scavenger and a DNase inhibitor. Furthermore, the organic mixture, and optionally after storage of the compo composition can comprise a metal salt as described herein, sition in a closed container (e.g., a capped tube, vial or well) where the aqueous mixture, the at least one alcohol Solvent or at a temperature from ambient temperature to about 40°C. for the aqueous organic mixture, or any combination thereof, can at least about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 comprise the metal salt (e.g., the metal salt can be added to the month, 2 months, 3 months, 6 months, 1 year, 1.5 years or 2 aqueous mixture, the at least one alcohol solvent or the aque years. In certain embodiments, the at least one alcohol Solvent ous organic mixture, or any combination thereof). in the composition comprises no more than about 10%, 5% or 0113. The composition can be re-hydrated by addition of 1% water by mass relative to the combined mass of water and an aqueous solution (e.g., water or an aqueous buffer) shortly the at least one alcohol solvent after removal of water from the before the composition is to be used in a biochemical reaction aqueous organic mixture, and optionally after storage of the (e.g., PCR) or an analysis (e.g., an immunoassay). composition in a closed container (e.g., a capped tube, vial or well) at a temperature from ambient temperature to about 40° C. for at least about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, Compositions Comprising a Biological Material in an 1 month, 2 months, 3 months, 6 months, 1 year, 1.5 years or Anhydrous, Ionic Organic Solvent 2 years. In some embodiments, the at least one alcohol solvent 0114. Other embodiments of the disclosure relate to com in the composition comprises no more than about 10%, 5% or positions comprising a biological material in a Substantially 1% water by mass relative to the combined mass of water and water-free, ionic organic solvent. In some embodiments, such the at least one alcohol solvent after removal of water from the a composition comprises: a polypeptide, a polynucleotide or aqueous organic mixture and after storage of the composition a biological sample, or any combination thereof, wherein the in a closed container (e.g., a capped tube, vial or well) at polypeptide comprises an enzyme that mediates a nucleic ambient temperature for at least about 3 months or 6 months. acid reaction, a polypeptide that regulates an enzyme, an 0110. The at least one alcohol solvent can comprise one or antibody, a polypeptide ligand of an antibody, a polypeptide more of any of the alcohol solvents described herein. Further aptamer, a protein or enzyme useful for detection, a toxin, a more, the polypeptide, the polynucleotide or the biological hormone, a cytokine, a polypeptide therapeutic or a vaccine, sample, or any combination thereof, can comprise any or a derivative thereof or any combination thereof; wherein polypeptide, any polynucleotide or any biological sample the polynucleotide comprises a polynucleotide used in a described herein. nucleic acid reaction, a catalytic polynucleotide, or a poly 0111. In certain embodiments, water is removed from the nucleotide that binds specifically to a target ligand, or a aqueous organic mixture by evaporation. In further embodi derivative thereof or any combination thereof; and wherein ments, removing water from the aqueous organic mixture to the biological sample comprises a biological fluid, a biologi produce the composition in a fluid State does not pass through cal Suspension, a fluid aspirate, blood, plasma, serum, lymph, an intermediate Solid State. In some embodiments, water is cerebrospinal fluid, gastric fluid, bile, perspiration, ocular removed from the aqueous organic mixture at ambient tem fluid, tears, oral fluid, sputum, saliva, a buccal sample, a tonsil perature or at reduced temperature but above the freezing sample, a nasal sample, mucus, a nasopharyngeal sample, point of the aqueous organic mixture or the at least one semen, urine, a vaginal sample, a cervical sample, a rectal alcohol solvent. In further embodiments, water is removed sample, a fecal sample, a wound or purulent sample, hair, a from the aqueous organic mixture at ambient pressure (e.g., at tissue, a tissue homogenate, cells, a cellular lysate, a tissue or about 1 atm) or at reduced pressure (e.g., at about 0.1 atm, 0.2 cell biopsy, skin cells, tumor or cancer cells, a microbe, a atm, atm, 0.4 atm, or 0.5 atm or greater). In certain embodi pathogen, a bacterium, a fungus, a protozoan or a virus, or any ments, water is removed from the aqueous organic mixture at combination thereof, and at least one ionic organic solvent ambient temperature and ambient pressure. In other embodi comprising an organic salt and an organic hydrogen bond ments, water is removed from the aqueous organic mixture at donor, wherein the composition is in a fluid State and is ambient temperature and reduced pressure. In yet other substantially free of water. US 2015/0267245 A1 Sep. 24, 2015

0115. In some embodiments, the at least one ionic organic 0.120. In certain embodiments, the at least one ionic Solvent in the composition comprises no more than about organic Solvent is a eutectic solvent. In further embodiments, 20%, 15%, 10%, 5%, 4%,3%,2%, 1% or 0.5% water by mass the at least one ionic organic solvent is a deep eutectic solvent. relative to the combined mass of water and the at least one I0121. In some embodiments, the molar ratio of the organic ionic organic solvent, e.g., after storage of the composition in salt to the organic hydrogen bond donor in the at least one a closed container (e.g., a capped tube, vial or well) at a ionic organic solvent is from about 5:1 to about 1:5, or from temperature from ambient temperature to about 40°C. for at about 4:1 to about 1:4, or from about 3:1 to about 1:3, or from least about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, about 2:1 to about 1:2, or from about 1.5:1 to about 1:1.5. In 2 months, 3 months, 6 months, 1 year, 1.5 years or 2 years. In certain embodiments, the molar ratio of the organic salt to the certain embodiments, the at least one organic hydrogen bond donor in the at least one ionic organic solvent is from about 1:1 to about 1:2, or is about 1:1, about 0116 ionic organic solvent in the composition comprises 1:1.5 or about 1:2. no more than about 10%, 5% or 1% water by mass relative to the combined mass of water and the at least one ionic organic 0.122 The organic salt of the at least one ionic organic Solvent, e.g., after storage of the composition in a closed Solvent can be any organic salt capable of forming a solvent container (e.g., a capped tube, vial or well) at a temperature with an organic hydrogen bond donor (e.g., by heating a from ambient temperature to about 40°C. for at least about 1 mixture of the organic salt and the organic hydrogen bond day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 donor). In certain embodiments, the organic salt of the at least months, 6 months, 1 year, 1.5 years or 2 years. In some one ionic organic solvent comprises one or more organic salts embodiments, the at least one ionic organic solvent in the selected from the group consisting of primary ammonium composition comprises no more than about 10%, 5% or 1% salts, secondary ammonium salts, tertiary ammonium salts, water by mass relative to the combined mass of water and the and quaternary ammonium salts. Examples of primary at least one ionic organic solvent after storage of the compo ammonium salts include without limitation methylammo sition in a closed container (e.g., a capped tube, vial or well) nium salts, ethylammonium salts, propylammonium salts, at ambient temperature for at least about 3 months or 6 butylammonium salts, 2-hydroxyethylammonium salts, months. 2-acetylethylammonium salts, 2-chloroethylammonium salts, 2-fluoroethylammonium salts, and benzylammonium 0117. In further embodiments, the at least one ionic salts. organic solventis Substantially soluble in water—e.g., at least (0123 Non-limiting examples of secondary ammonium about 50%, 60%, 70%, 80%, 90%, 95% or 99% of the at least salts include dimethylammonium salts, diethylammonium one ionic organic solvent by mass or Volume is soluble in salts, dipropylammonium salts, dibutylammonium salts, bis water. In certain embodiments, at least about 90%, 95% or (2-hydroxyethyl)-ammonium salts, dibenzylammonium 99% of the at least one ionic organic solvent by mass or salts, ethylmethylammonium salts, (2-hydroxyethyl)methyl Volume is soluble in water. In an embodiment, the at least one ammonium salts, (2-hydroxyethyl)ethylammonium salts, ionic organic solvent is miscible with water. Solubility of the (2-acetylethyl)methylammonium salts, (2-acetylethyl)ethy at least one ionic organic solvent in waterpromotes transfer of lammonium salts, (2-chloroethyl)methylammonium salts, a biological material from an aqueous medium to the at least (2-chloroethyl)ethyl-ammonium salts, (2-fluoroethyl)me one ionic organic solvent. thylammonium salts, (2-fluoroethyl)ethylammonium salts, 0118. In yet further embodiments, the at least one ionic benzylmethylammonium salts, benzylethylammonium salts, organic solvent has a boiling point Substantially greater than benzyl (2-hydroxyethyl)ammonium salts, benzyl (2-acetyl that of water—e.g., a boiling point at least or greater than ethyl)ammonium salts, benzyl (2-chloroethyl)ammonium about 125° C., 150° C., 1759 C., 200° C., 250° C. or 300° C. salts, and benzyl(2-fluoroethyl)ammonium salts. at a pressure of about 1 atm. In certain embodiments, the at 0.124 Non-limiting examples of tertiary ammonium salts least one ionic organic solvent has a boiling point at least or include trimethylammonium salts, triethylammonium salts, greater than about 150° C., 200° C. or 250° C. at a pressure of dimethylethylammonium salts, diethylmethylammonium about 1 atm. The at least one ionic organic solvent having a salts, (benzyl)(ethyl)methylammonium salts, (benzyl)dim boiling point greater than the boiling point of water allows for ethylammonium salts, (benzyl)diethylammonium salts, an aqueous mixture comprising a biological material to be (2-hydroxyethyl)dimethylammonium salts, (2-hydroxy mixed with at least one ionic organic solvent and for water to ethyl)diethylammonium salts, (2-acetylethyl)dimethylam be selectively removed (e.g., by evaporation) from the result monium salts, (2-acetylethyl)diethylammonium salts, ing aqueous organic mixture without Substantial loss of the at (2-chloroethyl)-dimethylammonium salts, (2-chloroethyl)di least one ionic organic solvent. ethylammonium salts, (2-fluoroethyl)dimethyl-ammonium 0119. In additional embodiments, the at least one ionic salts, (2-fluoroethyl)diethylammonium salts, (2-hydroxy organic solvent has a dynamic (or absolute) viscosity of no ethyl)(benzyl)methylammonium salts, (2-hydroxyethyl) more than about 2000, 1500, 1000, 500, 400, 300, 200, 100, (benzyl)ethylammonium salts, (2-acetylethyl) (benzyl)me 50 or 25 centipoise (cP) or mPa is at ambient temperature. In thylammonium salts, (2-acetylethyl)(benzyl) certain embodiments, the at least one ionic organic solvent ethylammonium salts, (2-chloroethyl)(benzyl) has a dynamic (or absolute) viscosity of no more than about methylammonium salts, (2-chloroethyl)(benzyl) 1000,500, 200, 100 or 50cP or mPasat ambient temperature. ethylammonium salts, (2-fluoroethyl)(benzyl) A lower dynamic (or absolute) viscosity of the at least one methylammonium salts, (2-fluoroethyl)(benzyl) ionic organic solvent allows for more facile handling of the ethylammonium salts, bis(2-hydroxyethyl) composition comprising the biological material and the at methylammonium salts, bis(2-hydroxyethyl) least one ionic organic solvent (e.g., using a pipette or other ethylammonium salts, and bis(2-hydroxyethyl) means of transferring the composition). benzylammonium salts. US 2015/0267245 A1 Sep. 24, 2015

0.125 Examples of quaternary ammonium salts include ylbutanamide, N-methylpentanamide, and N-methylbenza without limitation tetramethylammonium salts, tetraethylam mide. In an embodiment, the organic hydrogen bond donor monium salts, (2-hydroxyethyl)trimethylammonium (cho comprises acetamide. line) salts, (2-hydroxyethyl)triethylammonium salts, (2-acet I0131 Non-limiting examples of carboxylic acids include ylethyl)trimethylammonium salts, (2-acetylethyl)- adipic acid, benzoic acid, citric acid, ethylenediaminetet triethylammonium salts, (2-chloroethyl) raacetic acid, fumaric acid, maleic acid, malonic acid, oxalic trimethylammonium salts, (2-chloroethyl) acid, phenylacetic acid, phenylpropionic acid, propane-1,2, triethylammonium salts, (2-fluoroethyl)trimethylammonium 3-tricarboxylic acid (tricarballylic acid). Succinic acid, and salts, (2-fluoroethyl)triethylammonium salts, (benzyl)(dim tartaric acid. In certain embodiments, the organic hydrogen ethyl)(2-hydroxyethyl)ammonium salts, (benzyl)(dimethyl) bond donor comprises citric acid, malonic acid or oxalic acid, (2-acetylethyl)ammonium salts, (benzyl)(dimethyl)(2-chlo or any combination thereof. Examples of phenolic com roethyl)ammonium salts, (benzyl)(dimethyl)(2-fluoroethyl)- pounds include without limitation phenol and tyrosine. ammonium salts, (benzyl)(diethyl)(2-hydroxyethyl) I0132) Examples of acyclic alcohols include without limi ammonium salts, (benzyl)(diethyl)(2-acetylethyl) tation the linear and branched C2-C6 acyclic alcohols having ammonium salts, (benzyl)(diethyl)(2-chloroethyl) one or more hydroxyl groups and optionally comprising one ammonium salts, (benzyl)(diethyl)(2-fluoroethyl) or more halide atoms described herein. Examples of cyclic ammonium salts, bis(2-hydroxyethyl) dimethylammonium alcohols include without limitation ascorbic acid and the salts, bis(2-hydroxyethyl)-diethylammonium salts, bis(2- C3-C6 cyclic alcohols having one or more hydroxyl groups hydroxyethyl)(benzyl)(methyl)ammonium salts, bis(2-hy and three to six ring carbon atoms and optionally comprising droxyethyl)(benzyl)(ethyl)ammonium salts, (dimethyl) one or more halide atoms described herein. In certain embodi (ethyl)(2-hydroxyethyl)ammonium salts, (diethyl)(methyl) ments, the organic hydrogen bond donor comprises ethylene (2-hydroxyethyl)ammonium salts, (benzyl) glycol, 1,2-propanediol. 1,3-propanediol, glycerol, 1.2-bu trimethylammonium salts, and (benzyl)triethylammonium tanediol, 1,3-butanediol, 1,4-butanediol, 2,3-butanediol, 1.2, salts. 4-butanetriol or 1.5-pentanediol, or any combination thereof. 0126 The anion of the organic salt can be any anion In further embodiments, the organic hydrogen bond donor capable of interacting (e.g., complexing or hydrogen bond comprises ethylene glycol. 1,3-propanediol, glycerol or 1.2- ing) with an organic hydrogen bond donor. In some embodi butanediol, or any combination thereof. ments, the anion of the organic salt is a monovalent anion. In 0.133 Examples of ionic organic solvents include without certain embodiments, the anion of the organic salt (e.g., the limitation: choline chloride or ethylammonium chloride and primary ammonium salts, the secondary ammonium salts, the urea; choline chloride or ethylammonium chloride and aceta tertiary ammonium salts, and the quaternary ammonium salts mide; choline chloride or ethylammonium chloride and citric described herein) is fluoride, chloride, bromide, iodide, acid; choline chloride or ethylammonium chloride and mal acetate, formate, nitrate, perchlorate (ClO4-), phosphate, Sul onic acid; choline chloride or ethylammonium chloride and fate, tetrafluoroborate (BF4-), or thiocyanate (—SCN). oxalic acid; choline chloride or ethylammonium chloride and 0127. In some embodiments, the organic salt comprises a ethylene glycol; choline chloride or ethylammonium chloride quaternary ammonium salt. In certain embodiments, the and glycerol; and choline chloride or ethylammonium chlo organic salt comprises a (2-hydroxyethyl)trimethylammo ride and 1,2-butanediol. nium (choline) salt. In further embodiments, the organic salt I0134. The at least one ionic organic solvent can be pre comprises choline chloride or choline acetate. pared by any method known in the art. For example, the at 0128. The organic hydrogen bond donor of the at least one least one ionic organic solvent can be prepared by heating at ionic organic solvent can be any organic hydrogen bond elevated temperature (e.g., at about 50° C., 75° C. or 100° C. donor capable of forming a solvent with an organic salt, or or higher) a mixture comprising one or more organic salts and any organic hydrogenbond donor capable of interacting (e.g., one or more organic hydrogen bond donors with stirring to complexing or hydrogen bonding) with the anion of an produce a liquid (e.g., a homogeneous liquid). An organic salt organic salt. In certain embodiments, the organic hydrogen having a melting point above ambient temperature, and/oran bond donor of the at least one ionic organic solvent comprises organic hydrogen bond donor having a melting point above one or more organic hydrogen bond donors selected from the ambient temperature, can be used to prepare the at least one group consisting of urea compounds, thiourea compounds, ionic organic solvent. carbamates, amides, carboxylic acids, phenolic compounds, 0.135 The composition can comprise in the substantially acyclic alcohols, and cyclic alcohols. water-free ionic organic solvent any polypeptide, any poly 0129 Non-limiting examples of urea compounds and nucleotide or any biological sample, or any combination thiourea compounds include urea, N-methylurea, N,N'-dim thereof, described herein. To enhance, e.g., the stability and/ ethylurea, N,N-dimethylurea, N.N.N'-trimethylurea, thio or the solubility of the biological material in the substantially urea, N-methylthiourea, N,N'-dimethylthiourea, N,N-dim water-free ionic organic solvent, the composition can further ethylthiourea, and N.N.N'-trimethylthiourea. In an comprise a metal salt, and/or one or more Substances selected embodiment, the organic hydrogen bond donor comprises from the group consisting of reducing agents, antioxidants, lea. free radical scavengers, oxygen radical scavengers, hydroxyl 0130. Examples of carbamates include without limitation radical scavengers, singlet oxygen quenchers, hydroperoX methyl carbamate, ethyl carbamate, propyl carbamate, butyl ide-removing agents, protease inhibitors, nuclease inhibitors, carbamate, methyl N-methylcarbamate, ethyl N-methylcar ribonuclease (RNase) inhibitors, deoxyribonuclease (DNase) bamate, propyl N-methylcarbamate, and butyl N-methylcar inhibitors, metal chelators, preservatives, anti-microbials, bamate. Examples of amides include without limitation buffers (or buffering agents), detergents, and chaotropes, as acetamide, propanamide, butanamide, pentanamide, benza described herein. In certain embodiments, the composition mide, N-methylacetamide, methylpropanamide, N-meth comprises the biological material and: a protease inhibitor, an US 2015/0267245 A1 Sep. 24, 2015 oxygen radical scavenger; a metal chelator, a hydroxyl radical 99% of its function or activity after storage of the composition Scavenger, and an RNase inhibitor; or a hydroxyl radical comprising the biological material in a closed container (e.g., scavenger and a DNase inhibitor. a capped tube, vial or well) at a temperature from ambient 0136. The biological material is soluble in the substan temperature to about 40°C. for at least about 1 day, 3 days, 1 tially water-free ionic organic solvent, and thus may not need week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 to be re-dissolved for use in fluid-phase reactions or assays, months, 1 year, 1.5 years or 2 years. In certain embodiments, including nucleic acid amplification reactions based on PCR the biological material retains at least about 90% of its func and analytical and diagnostic assays, such as immunoassays. tion or activity after storage of the composition comprising In some embodiments, at least about 50%, 60%, 70%, 80%, the biological material in a closed container (e.g., a capped 90%. 95% or 99% of the biological material by mass is tube, vial or well) at ambient temperature for at least about 3 dissolved in the Substantially water-free ionic organic Sol months or 6 months. The function or activity of a biological vent, e.g., after storage of the composition comprising the material preserved in the substantially water-free ionic biological material in a closed container (e.g., a capped tube, organic solvent and tested at a particular time point can be vial or well) at a temperature from ambient temperature to compared to the function or activity of a positive control, e.g., about 40°C. for at least about 1 day, 3 days, 1 week, 2 weeks, the function or activity of the biological material prepared 3 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 1.5 under appropriate conditions (e.g., in an aqueous medium) years or 2 years. In certain embodiments, at least about 80% shortly before its use in the test protocol (e.g., test reaction or or 90% of the biological material by mass is dissolved in the test assay). Substantially water-free ionic organic solvent after storage of the composition comprising the biological material in a Method of Preserving a Biological Material in an Anhydrous, closed container (e.g., a capped tube, vial or well) at ambient Ionic Organic Solvent temperature for at least about 3 months or 6 months. 0.139. Additional embodiments of the disclosure relate to a 0.137 Furthermore, the biological material is stable (e.g., method of preserving a biological material in a Substantially retains its structural integrity) in the substantially water-free water-free, ionic organic solvent. In some embodiments, the ionic organic solvent at ambient temperature or higher, and method comprises: mixing an aqueous mixture comprising a thus does not need to be refrigerated or frozen during shipping polypeptide, a polynucleotide or a biological sample, or any or storage. In some embodiments, the biological material is combination thereof, with at least one ionic organic solvent to stable (e.g., retains its structural integrity) in the Substantially produce anaqueous organic mixture, wherein the polypeptide water-free ionic organic solvent after storage of the compo comprises an enzyme that mediates a nucleic acid reaction, a sition comprising the biological material in a closed container polypeptide that regulates an enzyme, an antibody, a polypep (e.g., a capped tube, vial or well) at a temperature from tide ligand of an antibody, a polypeptide aptamer, a protein or ambient temperature to about 40°C. for at least about 1 day, enzyme useful for detection, a toxin, a hormone, a cytokine, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 a polypeptide therapeutic or a vaccine, or a derivative thereof months, 6 months, 1 year, 1.5 years or 2 years. In further or any combination thereof; the polynucleotide comprises a embodiments, the biological material is resistant to irrevers polynucleotide used in a nucleic acid reaction, a catalytic ible hydrolytic damage, irreversible oxidative damage, and polynucleotide, or a polynucleotide that binds specifically to irreversible denaturation (e.g., irreversible unfolding or irre a target ligand, or a derivative thereof or any combination versible loss of secondary structure or tertiary structure) after thereof; the biological sample comprises a biological fluid, a storage of the composition comprising the biological material biological Suspension, a fluid aspirate, blood, plasma, serum, in a closed container (e.g., a capped tube, vial or well) at a lymph, cerebrospinal fluid, gastric fluid, bile, perspiration, temperature from ambient temperature to about 40°C. for at ocular fluid, tears, oral fluid, sputum, saliva, a buccal sample, least about 1 day, 3 days, week, 2 weeks, 3 weeks, 1 month, 2 a tonsil sample, a nasal sample, mucus, a nasopharyngeal months, 3 months, 6 months, 1 year, 1.5 years or 2 years. In sample, semen, urine, a vaginal sample, a cervical sample, a certain embodiments, the biological material is resistant to rectal sample, a fecal sample, a wound or purulent sample, irreversible hydrolytic damage, irreversible oxidative dam hair, a tissue, a tissue homogenate, cells, a cellular lysate, a age, and irreversible denaturation (e.g., irreversible unfolding tissue or cell biopsy, skin cells, tumor or cancer cells, a or irreversible loss of secondary structure or tertiary struc microbe, a pathogen, a bacterium, a fungus, a protozoan or a ture) after storage of the composition comprising the biologi virus, or any combination thereof, and the at least one ionic cal material in a closed container (e.g., a capped tube, vial or organic solvent comprises an organic salt and an organic well) at ambient temperature for at least about 3 months or 6 hydrogen bond donor, and removing water from the aqueous months. organic mixture to produce a composition comprising the 0.138. In addition, the biological material retains its func polypeptide, the polynucleotide or the biological sample, or tion or activity when it is preserved in the substantially water any combination thereof, and the at least one ionic organic free ionic organic solvent at ambient temperature or higher Solvent, wherein the composition is in a fluid state and is and is tested for its function or activity under appropriate substantially free of water. conditions (e.g., in an aqueous medium). In some embodi 0140. In some embodiments, the at least one ionic organic ments, the biological material retains its function or activity Solvent in the composition comprises no more than about after storage of the composition comprising the biological 20%, 15%, 10%, 5%, 4%,3%,2%, 1% or 0.5% water by mass material in a closed container (e.g., a capped tube, vial or relative to the combined mass of water and the at least one well) at a temperature from ambient temperature to about 40° ionic organic solvent after removal of water from the aqueous C. for at least about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, organic mixture, and optionally after storage of the compo 1 month, 2 months, 3 months, 6 months, 1 year, 1.5 years or sition in a closed container (e.g., a capped tube, vial or well) 2 years. In further embodiments, the biological material at a temperature from ambient temperature to about 40°C. for retains at least about 50%, 60%, 70%, 80%, 90%, 95% or at least about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 US 2015/0267245 A1 Sep. 24, 2015

month, 2 months, 3 months, 6 months, 1 year, 1.5 years or 2 ture, or any combination thereof, can comprise the metal salt years. In certain embodiments, the at least one ionic organic (e.g., the metal salt can be added to the aqueous mixture, the Solvent in the composition comprises no more than about at least one ionic organic Solvent or the aqueous organic 10%, 5% or 1% water by mass relative to the combined mass mixture, or any combination thereof). of water and the at least one ionic organic solvent after 0144. The composition can be re-hydrated by addition of removal of water from the aqueous organic mixture, and an aqueous solution (e.g., water oran aqueous buffer) shortly optionally after storage of the composition in a closed con before the composition is to be used in a biochemical reaction tainer (e.g., a capped tube, vial or well) at a temperature from (e.g., PCR) or an analysis (e.g., an immunoassay). ambient temperature to about 40°C. for at least about 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months, 1 year, 1.5 years or 2 years. In some Containers and Kits Comprising Biological Materials in embodiments, the at least one ionic organic solvent in the Anhydrous Fluid Media composition comprises no more than about 10%, 5% or 1% (0145. Further embodiments of the disclosure relate to con water by mass relative to the combined mass of water and the tainers and kits containing compositions comprising biologi at least one ionic organic solvent after removal of water from cal materials in substantially water-free fluid media. A con the aqueous organic mixture and after storage of the compo tainer can comprise any composition comprising a biological sition in a closed container (e.g., a capped tube, vial or well) material in a substantially water-free fluid medium described at ambient temperature for at least about 3 months or 6 herein. The container can be any vessel suitable for holding or months. storing a fluid composition. In certain embodiments, the con 0141. The at least one ionic organic solvent can comprise tainer is a tube, a vial, a well or a chamber (including a well or any ionic organic solvent described herein. Furthermore, the chamber in a cartridge). In further embodiments, the con polypeptide, the polynucleotide or the biological sample, or tainer is any vessel Suitable for keeping away moisture during any combination thereof, can comprise any polypeptide, any storage of a composition, such as a capped tube, Vial, well or polynucleotide or any biological sample described herein. chamber. A capped container can have any suitable cap. Such 0142. In certain embodiments, water is removed from the as a Snap-on cap or a screw cap. In certain embodiments, the aqueous organic mixture by evaporation. In further embodi container is a screw-cap tube or a screw-cap vial. A screw-cap ments, removing water from the aqueous organic mixture to tube or a screw-cap vial can have a gasket for improved produce the composition in a fluid State does not pass through sealing of the screw cap to the tube or the vial. an intermediate solid state. In some embodiments, water is 0146 A kit can contain one or more compositions com removed from the aqueous organic mixture at ambient tem prising a biological material in a Substantially water-free fluid perature or at reduced temperature but above the freezing medium described herein. The kit can contain one or more point of the aqueous organic mixture or the at least one ionic containers comprising one or more compositions, as organic solvent. In further embodiments, water is removed described herein. For example, a kit can contain reagents for from the aqueous organic mixture at ambient pressure (e.g., at performing a biochemical reaction (e.g., a nucleic acid ampli about 1 atm) or at reduced pressure (e.g., at about 0.1 atm, 0.2 fication reaction, such as PCR or RT-PCR) or an assay (e.g., atm, 0.3 atm, 0.4 atm, or 0.5 atm or greater). In certain an immunoassay, Such as an ELISA or a sandwich immunoas embodiments, water is removed from the aqueous organic say), wherein: the kit can include a container containing a mixture at ambient temperature and ambient pressure. In composition comprising all the reagents for performing the other embodiments, water is removed from the aqueous biochemical reaction or the assay; or the kit can include two organic mixture at ambient temperature and reduced pres or more containers, where each container contains a compo sure. In yet other embodiments, water is removed from the sition comprising one or more reagents for performing the aqueous organic mixture at reduced pressure and at reduced biochemical reaction or the assay as described herein, and the temperature but above the freezing point of the aqueous one or more reagents in each of the containers can be com organic mixture or the at least one ionic organic solvent. In bined prior to or at the time of their use in the biochemical further embodiments, water is removed from the aqueous reaction or the assay or can be used at different times in organic mixture at a relative humidity of no more than about performing the biochemical reaction or the assay as appro 60%, 50%, 40%, 30% or 20%. priate. 0143. The composition can be any composition compris ing a biological material in a Substantially water-free ionic 0147 If an antibody used in an immunoassay is conju organic solvent as described herein. For example, the com gated to a detection enzyme, the kit can further comprise a position can comprise one or more Substances as described Substrate with which the enzyme can react to produce a herein, where the aqueous mixture, the at least one ionic detectable signal (e.g., a color change in the Substrate). organic solvent or the aqueous organic mixture, or any com 0.148. As an example, a kit can comprise reagents for per bination thereof, can comprise the one or more substances forming PCR or RT-PCR, wherein: the kit can include a (e.g., the one or more substances can be added to the aqueous container containing a composition comprising all the mixture, the at least one ionic organic solvent or the aqueous reagents for performing PCR or RT-PCR; or the kit can organic mixture, or any combination thereof). In certain include two or more containers, where each container con embodiments, the one or more Substances comprise: (a) a tains a composition comprising one or more reagents for protease inhibitor; (b) an oxygen radical scavenger; (c) a performing PCR or RT-PCR as described herein (e.g., a DNA metal chelator, a hydroxyl radical scavenger, and an RNase polymerase in one container and at least one pair of forward inhibitor; or (d) a hydroxyl radical scavenger and a DNase and reverse primers in a separate container for PCR, or a inhibitor. Furthermore, the composition can comprise a metal reverse transcriptase and a DNA polymerase in one container salt as described herein, where the aqueous mixture, the at and at least one reverse transcription primer and at least one least one ionic organic solvent or the aqueous organic mix pair of forward and reverse primers in a separate container for US 2015/0267245 A1 Sep. 24, 2015

RT-PCR), and the one or more reagents in each of the con were added to the evaporated mixture in the PCR tube to a tainers can be combined prior to or at the time of their use in final volume of about 25 uL. Reverse transcription PCR was PCR or RT-PCR. performed according to the manufacturer's recommended 0149. As another example, a kit can comprise reagents for protocol. Production of the target 18S rRNA amplicon prod performing an ELISA or a sandwich immunoassay, wherein: uct having about 313 base pairs (bp) was analyzed by gel the kit can include two or more containers, each container electrophoresis using 2% agarose gel and UV visualization. containing a composition comprising one or more reagents Positive control was the Master Mix not treated with sucrose for performing the ELISA or the sandwich immunoassay as and not subjected to water removal, where the Master Mix described herein (e.g., a first antibody in one container and a was prepared by taking unmixed RT-PCR reagents out of a second antibody in a separate container for an ELISA or a -20° C. freezer and mixing them shortly before use at all sandwich immunoassay); and the one or more reagents in tested time points (untreated and unevaporated Master Mix), each of the containers can be used at different times in per with addition of the template RNA. Negative control was forming the ELISA or the Sandwich immunoassay as appro untreated and unevaporated Master Mix without addition of priate (e.g., the second antibody can be used Subsequent to the template RNA. use of the first antibody in the ELISA or the sandwich immu 0154 Preservation of the reagents was assessed by elec noassay). trophoresis of the RT-PCR products at time-0, about 7 days or 0150. A kit can further comprise water or an aqueous about 14 days of storage (FIG. 1). When RT-PCR was per Solution (e.g., an aqueous buffer) in a container (e.g., a vial, formed shortly after preparation of the evaporated mixture bottle or cartridge) for re-hydration of the biological material (t=0), the target 313 bp 18S rRNA product was produced if no in a composition for use, e.g., in a reaction (e.g., a PCR Sucrose or about 1.3 mg, 2.5 mg or 5 mg of Sucrose had been amplification reaction) or an assay (e.g., an analytical or added to the RT-PCR Master Mix. When RT-PCR was per diagnostic assay, Such as an immunoassay) which is per formed after about 7 days of storage of the evaporated mixture formed in an aqueous medium. In addition, a kit can comprise at ambient temperature (t–7 days), addition of no Sucrose to a desiccant for promoting preservation of the biological mate the Master Mix yielded a much lower amount of the target 313 rial in a Substantially anhydrous state. Non-limiting examples bp product compared to addition of about 1.3 mg, 2.5 mg or 5 of desiccants include activated alumina, aerogel, silica gel. mg of sucrose to the Master Mix. When RT-PCR was per benzophenone, calcium chloride, calcium sulfate, cobalt formed after about 14 days of storage of the evaporated mix chloride, copper sulfate, lithium chloride, lithium bromide, ture at ambient temperature (t=14 days), addition of no magnesium chloride, magnesium perchlorate, magnesium sucrose or about 1.3 mg of sucrose to the Master Mix resulted Sulfate, potassium carbonate, sodium chlorate, Sodium chlo in no detectable target 313 bp product, whereas addition of ride, sodium hydroxide, Sodium sulfate. Sucrose, clay (e.g., about 2.5 mg or 5 mg of sucrose to the Master Mix produced bentonite clay and montmorillonite clay), and molecular the target 313 bp product. sieves. Moreover, a kit can comprise instruction for storing and using a composition, and optionally instruction for using Example 2 water or an aqueous solution (e.g., an aqueous buffer) to re-hydrate the biological material in a composition. Preservation of RT-PCR Reagents in Gycerol for Avian Flu Virus RNA Analysis EXAMPLES (O155 An undiluted volume (about 25 t L) of the RT-PCR 0151. The following examples are intended only to illus Master Mix of the TaqMan R. Avian Influenza Virus (AIV-M) trate the disclosure. Other procedures, methodologies, tech Reagents (Life Technologies) for analysis of AIV matrix niques, conditions and reagents may alternatively be used as (AIV-M) RNA was placed in a well of a 96-well optical PCR appropriate. plate. The volume of Master Mix contained some amount (e.g., about 0.5-4 uL) of glycerol. Varying amounts of Sucrose Example 1 (0 mg. or about 1.3 mg or 2.5 mg) were added to the Master Mix. Water was removed from the resulting mixture by Preservation of RT-PCR Reagents in Gycerol for evaporation at reduced pressure (about 0.2 atm) and ambient Human 18S rRNA Analysis temperature overnight to yield a fluid mixture having a Vol ume of about 4 LL. The evaporated mixture containing the 0152. An undiluted volume (about 25 uL) of the Master RT-PCR reagents and optionally sucrose in glycerol was Mix of the Ag-Path IDTM. One-Step RT-PCR Kit (Life Tech stored in the well capped with a Snap-on cap at ambient nologies) for analysis of human 18S ribosomal RNA (rRNA) temperature (about 25°C.) for varying periods of time (about was placed in a PCR tube. The volume of Master Mix con 0 day or 7 days). The PCR plate was kept in the dark during tained some amount (e.g., about 0.5-4 uL) of glycerol. Vary storage to protect a dye-labeled probe in the RT-PCR reagents ing amounts of Sucrose (0 mg, or about 1.3 mg, 2.5 mg or 5 from light. mg) were added to the Master Mix. Water was removed from 0156 For analysis of avian influenza virus matrix RNA, the resulting mixture by evaporation at reduced pressure deionized nuclease-free water and template RNA (about 10 (about 0.2 atm) and ambient temperature overnight to yield a ng of AIV genomic RNA) were added to the evaporated fluid mixture having a volume of about 4 LL. The evaporated mixture in the well to a final volume of about 25uL. Reverse mixture containing the RT-PCR reagents and optionally transcription PCR was performed according to the manufac sucrose in glycerol was stored in the PCR tube capped with a turer's recommended protocol. Production of the target Snap-on cap at ambient temperature (about 25°C.) for vary amplicon product of the matrix (M) gene of AIV was ana ing periods of time (about 0 day, 7 days or 14 days). lyzed using an Applied Biosystems 7500 Fast Real-Time 0153. For analysis of human 18S rRNA, nuclease-free PCR System (Life Technologies), which generated Ct values water and template RNA (about 10 ng of HeLa total RNA) when a fluorescent signal reached a level detectable by the US 2015/0267245 A1 Sep. 24, 2015

real-time PCR instrument. Positive control was the Master erol was stored in the PCR tube capped with a snap-on cap at Mix not treated with sucrose and not subjected to water ambient temperature (about 25°C.) for varying periods of removal, where the Master Mix was prepared by taking time (about 0 day or 7 days). The PCR tube was kept in the unmixed RT-PCR reagents out of a -20°C. freezer and mix dark during storage to protect the dye-labeled primers from ing them shortly before use at all tested time points untreated light. and unevaporated Master Mix), with addition of the template 0159 For analysis of the 13 CODIS STR loci, Penta D, RNA. Negative control was untreated and unevaporated Mas Penta E and amelogenin, nuclease-free water and template ter Mix without addition of the template RNA. DNA (about 1 ng of human genomic DNA) were added to the 0157 Table 1 shows results when reverse transcription evaporated mixture in the PCR tube to a final volume of about PCR was performed at time=0 or after about 7 days of storage. 20 uL. Multiplex PCR was performed according to the manu When RT-PCR was performed shortly after preparation of the facturer's recommended protocol. Production of the ampli evaporated mixture (t=0), addition of no sucrose to the RT con products of the 16 loci was analyzed using an Applied PCR Master Mix yielded a slightly lower amount of the target Biosystems 3130 Genetic Analyzer instrument (Life Tech matrix product compared to addition of about 1.3 mg or 2.5 nologies). Positive control was the Master Mix not treated mg of sucrose to the Master Mix. When RT-PCR was per with sucrose and not subjected to water removal, where the formed after about 7 days of storage of the evaporated mixture Master Mix was prepared by taking unmixed PCR reagents at ambient temperature (t=7 days), addition of no Sucrose to out of a -20°C. freezer and mixing them shortly before use at the Master Mix resulted in no detectable target matrix prod all tested time points (untreated and unevaporated Master uct, while addition of about 1.3 mg of sucrose to the Master Mix), with addition of the template DNA. Negative control Mix produced a lower amount of the target matrix product was untreated and unevaporated Master Mix without addition than addition of about 2.5 mg of sucrose to the Master Mix. of the template DNA. 0160 Electropherograms generated by amplicon product TABLE 1. analysis of multiplex PCR products performed at time=0 or after about 7 days of storage are shown in FIGS. 2 and 3. Results of RT-PCR respectively. When PCR was performed shortly after prepa ration of the evaporated mixture (t=0), addition of no sucrose time = 0 time = 7 days or about 1 mg or 2 mg of sucrose to the PCR Master Mix A Ave. Ct A Ave. Ct yielded similar amounts of the amplicon products of the 16 Ave. (vs. pos. Ave. (vs. pos. loci (FIG.2). When PCR was performed after about 7 days of Ct Ct control) Ct Ct control) storage of the evaporated mixture at ambient temperature (t=7 Positive 25.02 25.33 24.92 24.89 days), addition of no sucrose to the Master Mix resulted in control 25.64 24.86 lower amounts of the amplicon products of the 16 loci com (wet) No Sucrose 27.00 26.89 1.56 >>35 pared to addition of about 1 mg or 2 mg of Sucrose to the (dry) 26.77 >>35 Master Mix (FIG.3). 1.3 mg 26.07 25.91 O.S8 36.22 34.34 9.45 SUCOS 25.75 32.47 Example 4 (dry) 2.5 mg 25.85 25.63 O.30 27.72 27.04 2.15 SUCOS 25.40 26.37 Preservation of Human Serum Solids in Glycerol (dry) 0.161 An aqueous solution containing varying amounts of Negative >>35 >>35 control glycerol (0 mg, or about 2.5 mg, 5 mg, 7.5 mg or 10 mg) and (wet) an aqueous solution containing varying amounts of Sucrose (0 mg, or about 5 mg, 7.5 mg or 10 mg) were added to a Volume (about 50 uL) of human serum in natural serum fluid in a tube. The volume of human serum contained about 7.5 mg of serum Example 3 solids (non-volatile components of serum). Water was removed from the resulting mixture by evaporation at reduced Preservation of PCR Reagents in Gycerol for pressure (about 0.2 atm) and ambient temperature for about STR-Based Human Identification two days. 0158 An undiluted volume (about 20 uL) of the Master 0162 FIG. 4 shows the approximate volume of evaporated Mix of the PowerPlex(R) 16 HS System (Promega) for human mixtures containing about 7.5 mg of human serum Solids and identification was placed in a PCR tube. The volume of Mas varying amounts of glycerol and Sucrose, after evaporation of ter Mix contained some amount (e.g., about 0.5-4 uL) of water at about 0.2 atm and ambient temperature for about two glycerol. The Master Mix contained 16 different pairs of days. The five evaporated mixtures containing human serum forward and reverse primers for amplifying all 13 current Solids and Sucrose in glycerol were clear and showed no CODIS STR loci, as well as Penta D, Penta E and ameloge apparent flocculation or precipitation of serum Solids. nin. Each primer was labeled with a fluorescent dye, and a total of three different fluorescent dyes were used to label the Example 5 16 different pairs of forward and reverse primers. Varying amounts of sucrose (0 mg, or about 1 mg or 2 mg) were added Preservation of RT-PCR Reagents in Glycerol or to the Master Mix. Water was removed from the resulting Glycerol/1,3-Propanediol for Human 18S rRNA mixture by evaporation at reduced pressure (about 0.2 atm) Analysis and ambient temperature overnight to yield a fluid mixture 0163 An undiluted volume (about 25 uL) of the Master having a Volume of about 4 LL. The evaporated mixture Mix of the Ag-Path IDTM. One-Step RT-PCR Kit (Life Tech containing the PCR reagents and optionally Sucrose in glyc nologies) for analysis of human 18S ribosomal RNA (rRNA) US 2015/0267245 A1 Sep. 24, 2015 20 was placed in a PCR tube. The volume of Master Mix con What is claimed is: tained some amount (e.g., about 0.5-4 uL) of glycerol. 1. A composition comprising: Sucrose (about 2.5 mg), and glycerol (about 1 LL) or 1.3- a) biological material, comprising a polypeptide, a poly propanediol (about 1 LL), were added to the Master Mix. nucleotide or a biological sample, or any combination Water was removed from the resulting mixture by evapora thereof, and tion at reduced pressure (about 0.2 atm) and ambient tem b) at least one solvent, wherein said solvent or solvents are perature overnight to yield a fluid mixture having a Volume of (i) at least one alcohol solvent selected from the group about 8 t L. After adding mineral oil (about 1 uL), 1% Tri consisting of linear and branched C2-C6 acyclic alco ton R-X (about 1 uL of a stock solution diluted 100-fold with hols having one or more hydroxyl groups and C3-C6 distilled water) or 1% Tween(R)-20 (about 1 uL of a stock cyclic alcohols having one or more hydroxyl groups solution diluted 100-fold with distilled water), or no addi and three to six ring carbonatoms, wherein the acyclic tional substance, to the fluid mixture, the mixture was sub alcohols and the cyclic alcohols optionally comprise jected to reduced pressure (about 0.2 atm) at ambient tem one or more halide atoms; or perature for about two additional hours. The evaporated (ii) at least one ionic organic solvent comprising an mixture containing the RT-PCR reagents and Sucrose, and organic salt and an organic hydrogen bond donor, optionally mineral oil or a detergent, in glycerol or glycerol/ wherein the composition is in a fluid state and is substantially 1,3-propanediol was rehydrated for immediate testing or was free of water. stored in the PCR tube capped with a snap-on cap at ambient 2. The composition of claim 1, wherein the at least one temperature (about 25°C.) for about 7 days before rehydra alcohol solvent is substantially soluble in water; has a boiling tion and testing. point Substantially greater than that of water, and comprises no more than about 10% water by mass. 0164. For analysis of human 18S rRNA, nuclease-free 3. The composition of claim 1, wherein the at least one water and template RNA (about 10 ng of HeLa total RNA) alcohol solvent comprises ethylene glycol, 1.2-propanediol. were added to the evaporated mixture in the PCR tube to a 1,3-propanediol, glycerol. 1.2-butanediol. 1,3-butanediol. final volume of about 25 uL. Reverse transcription PCR was 1,4-butanediol. 2,3-butanediol, 1,2,4-butanetriol or 1.5-pen performed according to the manufacturer's recommended tanediol, or any combination thereof. protocol. Production of the target 18S rRNA amplicon prod 4. The composition of claim 1, wherein said biological uct having about 313 base pairs (bp) was analyzed by gel material comprises one or more reagents for performing PCR, electrophoresis using 2% agarose gel and ethidium bromide wherein: for UV visualization. the reagents for performing PCR comprise a DNA poly 0.165 Preservation of the reagents was assessed by elec merase and at least one pair of a forward primer and a trophoresis of the RT-PCR products generated at time=0 or reverse primer for amplifying at least one nucleic acid after about 7 days of storage in Sucrose and glycerol or 1.3- locus; and propanediol (“1.3-PD), and optionally mineral oil or a deter the at least one pair of forward primer and reverse primer gent, added to the Master Mix (FIGS. 5A and 5B). For the optionally is labeled with a dye. “No overlay tests, no mineral oil and no detergent had been 5. The composition of claim 4, wherein the at least one pair added to the Master Mix. For the "Mineral oil” tests, mineral of forward and reverse primers comprises at least 5 different oil, but no detergent, had been added to the Master Mix, and pairs of forward and reverse primers for amplifying at least 5 the mineral oil was removed before RT-PCR was conducted. different short tandem repeat (STR) loci utilized in a forensic For the “1% Triton-X' tests, 1% Triton(R)-X, but no mineral database, and wherein each of the at least 5 different pairs of oil, had been added to the Master Mix. For the “1% Tween forward and reverse primers optionally is labeled with a dye. 20' tests, 1% Tween R-20, but no mineral oil, had been added 6. The composition of claim 1, wherein said biological to the Master Mix. These results show that RT-PCR generated material comprises one or more reagents for performing comparable levels of the target 313 bp 18S rRNA amplicon reverse transcription PCR (RT-PCR), wherein: product at time-0 or about 7 days after sucrose and glycerol the reagents for performing RT-PCR comprise a reverse or 1,3-propanediol, and optionally mineral oil or a detergent, transcriptase, a DNA polymerase, at least one reverse had been added to the Master Mix. transcription primer for reverse transcribing at least one polyribonucleotide to produce at least one polydeoxyri 0166 Therefore, the present invention is well adapted to bonucleotide complementary to the at least one polyri attain the ends and advantages mentioned as well as those that bonucleotide, and at least one pair of a forward primer are inherent therein. The particular embodiments disclosed and a reverse primer for amplifying the at least one above are illustrative only, as the present invention may be complementary polydeoxyribonucleotide; and modified and practiced in different but equivalent manners wherein the at least one reverse transcription primer option apparent to those skilled in the art having the benefit of the ally is labeled with a dye and the at least one pair of forward teachings herein. Furthermore, no limitations are intended to primer and reverse primer optionally is labeled with a dye. the details of construction or design herein shown, other than 7. The composition of claim 1, wherein said biological as described in the claims below. It is therefore evident that material comprises one or more reagents for performing an the particular illustrative embodiments disclosed above may immunoassay, wherein the one or more reagents for perform be altered, combined, or modified and all such variations are ing an immunoassay comprise an antibody that is specific for considered within the scope and spirit of the present inven a target antigen or analyte; the antibody is labeled with a dye tion. or is conjugated to a detection protein or enzyme; and the 0167. The invention being thus described, it will be obvi antibody optionally is bound to a solid substrate. ous that the same may be varied in many ways. Such varia 8. The composition of claim 7, further comprising one or tions are not to be regarded as a departure from the spirit and more reagents for performing a sandwich immunoassay, Scope of the inventions. wherein the reagents for performing a sandwich immunoas US 2015/0267245 A1 Sep. 24, 2015

say comprise a first antibody that is specific for a target 14. The method of claim 13, wherein the at least one antigen or analyte and a second antibody that is specific for Solvent is Substantially soluble in water, has a boiling point the target antigen or analyte; said second antibody is labeled Substantially greater than that of water; and comprises no with a dye or is conjugated to a detection protein or enzyme; more than about 10% water by mass. and the first antibody optionally is bound to a solid substrate. 15. The method of claim 13, wherein said removing water 9. The composition of claim 1, wherein said biological from the aqueous organic mixture comprises evaporation. material comprises: 16. The method of claim 13, wherein said removing water a) whole or fractionated animal blood; from the aqueous organic mixture does not produce, include, b) whole or fractionated animal plasma; or or involve an intermediate solid state. c) whole or fractionated animal serum. 17. The composition of claim 1, wherein the at least one 10. The composition of claim 1, wherein said biological ionic organic solvent is a eutectic solvent. material comprises animal cells, mammalian cells, human 18. The composition of claim 1, wherein the molar ratio of cells, plant cells, microbial cells, pathogenic cells, bacterial the organic salt to the organic hydrogen bond donor in the at cells, fungal cells, protozoan cells or viral particles, or any least one ionic organic solvent is from about 1:1 to about 1:2. combination thereof, or lysates or extracts thereof. 19. The composition of claim 1, wherein the organic salt of 11. The composition of claim 1, further comprising one or the at least one ionic organic solvent comprises one or more more Substances selected from the group consisting of reduc organic salts selected from the group consisting of primary ing agents, antioxidants, free radical Scavengers, oxygen radi ammonium salts, secondary ammonium salts, tertiary ammo cal scavengers, hydroxyl radical scavengers, singlet oxygen nium salts, and quaternary ammonium salts. quenchers, hydroperoxide-removing agents, protease inhibi 20. The composition of claim 19, wherein the organic salt tors, nuclease inhibitors, ribonuclease (RNase) inhibitors, comprises a choline salt. deoxyribonuclease (DNase) inhibitors, metal chelators, pre 21. The composition of claim 1, wherein the organic hydro servatives, anti-microbials, buffers (or buffering agents), gen bond donor of the at least one ionic organic solvent detergents, chaotropes, M' salts, and M" salts. comprises one or more organic hydrogen bond donors 12. The composition of claim 1, wherein said biological selected from the group consisting of urea compounds, thio material is stable, retains its function, and/or retains its activ urea compounds, carbamates, amides, carboxylic acids, phe ity after storage at ambient temperature for at least about 2 nolic compounds, acyclic alcohols, and cyclic alcohols. weeks or 1 month. 22. The composition of claim 21, wherein the organic 13. A method of preserving a biological material, compris hydrogen bond donor comprises urea, acetamide, citric acid, ing: malonic acid, oxalic acid, ethylene glycol, 1.2-propanediol. a) mixing an aqueous mixture comprising the biological 1,3-propanediol, glycerol. 1.2-butanediol. 1,3-butanediol. material with at least one solvent, wherein said solvent or solvents are 1,4-butanediol. 2,3-butanediol, 1,2,4-butanetriol or 1.5-pen (i) at least one alcohol Solvent selected from the group tanediol, or any combination thereof. consisting of linear and branched C2-C6 acyclic alco 23. A container comprising the composition of claim 1. hols having one or more hydroxyl groups and C3-C6 24. A kit for preserving biological material comprising: cyclic alcohols having one or more hydroxyl groups a closable container for storage; and and three to six ring carbonatoms, wherein the acyclic at least one solvent, wherein said solvent or solvents are (i) alcohols and the cyclic alcohols optionally comprise at least one alcohol solvent selected from the group one or more halide atoms; or consisting of linear and branched C2-C6 acyclic alco (ii) at least one ionic organic solvent comprising an hols having one or more hydroxyl groups and C3-C6 organic salt and an organic hydrogen bond donor, and cyclic alcohols having one or more hydroxyl groups and b) removing water from the aqueous organic mixture to three to six ring carbon atoms, wherein the acyclic alco produce a composition comprising the polypeptide, the hols and the cyclic alcohols optionally comprise one or polynucleotide or the biological sample, or any combi more halide atoms; or (ii) at least one ionic organic nation thereof, and the at least one alcohol solvent, Solvent comprising an organic salt and an organic hydro wherein the composition is in a fluid state and is substantially gen bond donor. free of water.