Analytical Methods Accepted Manuscript

Total Page:16

File Type:pdf, Size:1020Kb

Analytical Methods Accepted Manuscript Analytical Methods Accepted Manuscript This is an Accepted Manuscript, which has been through the Royal Society of Chemistry peer review process and has been accepted for publication. Accepted Manuscripts are published online shortly after acceptance, before technical editing, formatting and proof reading. Using this free service, authors can make their results available to the community, in citable form, before we publish the edited article. We will replace this Accepted Manuscript with the edited and formatted Advance Article as soon as it is available. You can find more information about Accepted Manuscripts in the Information for Authors. Please note that technical editing may introduce minor changes to the text and/or graphics, which may alter content. The journal’s standard Terms & Conditions and the Ethical guidelines still apply. In no event shall the Royal Society of Chemistry be held responsible for any errors or omissions in this Accepted Manuscript or any consequences arising from the use of any information it contains. www.rsc.org/methods Page 1 of 54 Analytical Methods 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Manuscript 15 16 17 18 19 20 Accepted 21 22 CEC 23 CIEF 24 CGE 25 CZE 26 Methods 27 28 29 30 31 32 33 Analytical 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 Analytical Methods Page 2 of 54 1 2 3 Review 4 5 6 Recent advances in the analysis of therapeutic proteins by capillary and microchip electrophoresis 7 1,3 2,3 1,2,3 8 Jessica S. Creamer, Nathan J. Oborny, Susan M. Lunte 9 10 1Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA 11 12 2Department of Bioengineering, University of Kansas, Lawrence, KS, USA 13 14 3Ralph N. Adams Institute for Bioanalytical Chemistry, University of Kansas, Lawrence, KS, USA 15 16 17 18 19 Susan M. Lunte, Ph.D. 20 21 Ralph N. Adams Professor of Chemistry and Pharmaceutical Chemistry Manuscript 22 23 Ralph N. Adams Institute for Bioanalytical Chemistry 24 25 2030 Becker Dr. Lawrence, KS, 66044 26 27 [email protected] 28 29 30 Fax: 785-864-1916 31 Accepted 32 List of abbreviations in alphabetical order: 33 + 34 1-vinyl-3-octylimidazolium (ViOcIm ), 8-aminopyrene-1,3,6-trisulfonic acid (APTS), background 35 − electrolyte (BGE), bis-trifluoromethanesulfonylimide (NTf 2 ), bovine serum albumin (BSA), capillary 36 37 electrochromatography (CEC), capillary electrophoresis (CE), capillary gel electrophoresis (CGE), capillary 38 isoelectric focusing (CIEF), capillary isotachophoresis (CITP), capillary zone electrophoresis (CZE), 39 diazoresin (DR), electroosmotic flow (EOF), electrospray ionization (ESI), erythropoietin (EPO), galactose- 40 Methods α-1,3-galactose (α1,3-Gal), gold nanoparticles (AuNP), graphene (G), graphene oxide (GO), hydrophilic 41 42 interaction chromatography (HILIC), hydroxyproplymethylcellulose (HPMC), imaging capillary isoelectric 43 focusing (iCIEF), ionic liquid (IL), isoelectric point (pI), laser-induced fluorescence (LIF), liquid 44 chromatography (LC), mass spectrometry (MS), matrix-assisted laser desorption ionization (MALDI), 45 46 microchip electrophoresis (ME), microchip gel electrophoresis (MGE), microchip isoelectric focusing 47 (MIEF), molecular weight (MW), monoclonal antibodies (mAbs), nanoparticles (NP), N- 48 glycolylneuraminic acid (Neu5Gc), N-methyl-2-pyrrolidonium methyl sulfonate ([NMP] +CH SO – ), open- 49 3 3 50 tubular capillary electrochromatography (OTCEC), pentaerythritol (PETA), phospholipid bilayers (PLB), Analytical 51 polyamidoamine-grafted silica nanoparticles (PAMAM-SNP), polybrene (PB), polyethylene glycol (PEG), 52 polyvinyl alcohol (PVA), post translational modifications (PTMs), pseudostationary phase (PSP), 53 54 quaternized celluoses (QC), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), silica 55 nanoparticles (SNP), sodium dodecyl sulfate (SDS), stabilized phospholipid bilayer (SPB), sulfobutyl ether 56 β-cyclodextrins (SBE β-CD) 57 58 59 60 1 Page 3 of 54 Analytical Methods 1 2 3 Abstract 4 5 6 The development of therapeutic proteins and peptides is an expensive and time-intensive process. 7 8 Biologics, which have become a multi-billion dollar industry, are chemically complex products that 9 10 require constant observation during each stage of development and production. Post-translational 11 12 13 modifications along with chemical and physical degradation from oxidation, deamidation, and 14 15 aggregation, lead to high levels of heterogeneity that affect drug quality and efficacy. The various 16 17 separation modes of capillary electrophoresis (CE) are commonly utilized to perform quality control and 18 19 assess protein heterogeneity. This review attempts to highlight the most recent developments and 20 21 Manuscript 22 applications of CE separation techniques for the characterization of protein and peptide therapeutics by 23 24 focusing on papers accepted for publication in the in the two-year period between January 2012 and 25 26 December 2013. The separation principles and technological advances of CE, capillary gel 27 28 29 electrophoresis, capillary isoelectric focusing, capillary electrochromatography and CE-mass 30 31 spectrometry are discussed, along with exciting new applications of these techniques to relevant Accepted 32 33 pharmaceutical issues. Also included is a small selection of papers on microchip electrophoresis to show 34 35 36 the direction this field is moving with regards to the development of inexpensive and portable analysis 37 38 systems for on-site, high-throughput analysis. 39 40 Methods 41 42 43 44 45 46 47 48 49 50 51 Analytical 52 53 54 55 56 57 58 59 60 2 Analytical Methods Page 4 of 54 1 2 3 1. Introduction 4 5 6 The characterization of protein therapeutics presents a unique analytical challenge due to the inherent 7 8 heterogeneity of recombinant protein expression. Even small changes in the manufacturing process can 9 10 lead to vastly different active pharmaceutical ingredients. Additionally, numerous physical and chemical 11 12 13 degradation pathways can occur during manufacturing and storage that compromise protein integrity, 14 15 leading to a potentially harmful, unstable product [1]. Thorough characterization of protein therapeutics 16 17 is necessary at every step of the research and development process, from drug discovery to lot release. 18 19 Due to the potential complexity of product degradation during preformulation and formulation 20 21 Manuscript 22 studies, additional separation techniques are needed to complement the more widely used column 23 24 liquid chromatography (LC) methods. To address this issue, capillary electrophoresis (CE) has become a 25 26 popular choice for the separation and analysis of therapeutic proteins and peptides. 27 28 29 CE provides several distinct advantages over LC. First, due to the faster separation times and the use 30 31 of multi-capillary arrays, hundreds of samples can be processed by CE per day. Second, CE is capable of Accepted 32 33 achieving very high efficiency separations due to the low diffusion coefficients of biomolecules. Lastly, 34 35 36 the small dimensions of the capillary and the low sample volume requirements keep reagent and 37 38 analyte use to a minimum, reducing the cost-per-test. The benefits of CE for the analysis of therapeutic 39 40 peptides and proteins have been addressed in several excellent reviews to date [2-5]. Methods 41 42 This review is aimed at highlighting the advances made in the field of CE therapeutic protein analysis 43 44 45 during 2012 and 2013 by expanding on a paper that was recently published by Zhao et al. [5]. Following 46 47 brief descriptions of the working principles of the different CE separation and detection methods, the 48 49 recent technological improvements and novel applications are discussed. Two additional sections have 50 51 Analytical 52 been included to further explore the use of CE for the determination of protein glycosylation and the 53 54 comparison of biosimilars. Finally, a brief introduction into microfluidic approaches to protein analysis is 55 56 given. Microchip electrophoresis (ME) has the additional advantages of increased speed, high- 57 58 59 60 3 Page 5 of 54 Analytical Methods 1 2 3 throughput capabilities, and portability for on-site analyses. Tables are presented in each section to 4 5 6 highlight the relevant CE and ME application-based citations. 7 8 2. Techniques 9 10 Historically, capillary zone electrophoresis (CZE) has been the most commonly employed form of CE. Yet, 11 12 13 the principles of electrophoretic separations and the benefits of capillary-based techniques are 14 15 applicable to other CE separation modes as well. Protein analysis based on size can be accomplished by 16 17 capillary gel electrophoresis (CGE), capillary isoelectric focusing (CIEF) can be used to determine 18 19 isoelectric points and charge heterogeneity, and capillary electrochromatography (CEC), which combines 20 21 Manuscript 22 the high efficiency electrophoretic separation with chromatographic retention, can be used for more 23 24 selective separations and analysis of neutral species. Depending on the properties of the analyte and 25 26 requirements of the assay, each of these separation modes can be coupled to a number of detection 27 28 29 methods such as UV-Vis absorbance, laser-induced fluorescence (LIF), and mass spectrometry (MS). 30 31 2.1 Capillary zone electrophoresis Accepted 32 33 Of the electrophoresis-based separation techniques, CZE is most frequently used for the analysis of
Recommended publications
  • Enabling Sweat-Based Biosensors: Solving the Problem of Low
    Enabling sweat-based biosensors: Solving the problem of low biomarker concentration in sweat A dissertation submitted to the Graduate School of the University of Cincinnati in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Biomedical Engineering of the College of Engineering & Applied Science by Andrew J. Jajack B.S., Biology, Wittenberg University, 2014 Committee Chairs: Jason C. Heikenfeld, Ph.D. and Chia-Ying Lin, Ph.D. Abstract Non-invasive, sweat biosensing will enable the development of an entirely new class of wearable devices capable of assessing health on a minute-to-minute basis. Every aspect of healthcare stands to benefit: prevention (activity tracking, stress-level monitoring, over-exertion alerting, dehydration warning), diagnosis (early-detection, new diagnostic techniques), and management (glucose tracking, drug-dose monitoring). Currently, blood is the gold standard for measuring the level of most biomarkers in the body. Unlike blood, sweat can be measured outside of the body with little inconvenience. While some biomarkers are produced in the sweat gland itself, most are produced elsewhere and must diffuse into sweat. These biomarkers come directly from blood or interstitial fluid which surrounds the sweat gland. However, a two-cell thick epithelium acts as barrier and dilutes most biomarkers in sweat. As a result, many biomarkers that would be useful to monitor are diluted in sweat to concentrations below what can be detected by current biosensors. This is a core challenge that must be overcome before the advantages of sweat biosensing can be fully realized. The objective of this dissertation is to develop methods of concentrating biomarkers in sweat to bring them into range of available biosensors.
    [Show full text]
  • Egg Consumption and Human Health
    nutrients Egg Consumption and Human Health Edited by Maria Luz Fernandez Printed Edition of the Special Issue Published in Nutrients www.mdpi.com/journal/nutrients Egg Consumption and Human Health Special Issue Editor Maria Luz Fernandez MDPI • Basel • Beijing • Wuhan • Barcelona • Belgrade Special Issue Editor Maria Luz Fernandez University of Connecticut USA Editorial Office MDPI AG St. Alban-Anlage 66 Basel, Switzerland This edition is a reprint of the Special Issue published online in the open access journal Nutrients (ISSN 2072-6643) in 2015–2016 (available at: http://www.mdpi.com/journal/nutrients/special issues/egg-consumption-human-health). For citation purposes, cite each article independently as indicated on the article page online and as indicated below: Lastname, F.M.; Lastname, F.M. Article title. Journal Name. Year. Article number, page range. First Edition 2018 ISBN 978-3-03842-666-0 (Pbk) ISBN 978-3-03842-667-7 (PDF) Articles in this volume are Open Access and distributed under the Creative Commons Attribution (CC BY) license, which allows users to download, copy and build upon published articles even for commercial purposes, as long as the author and publisher are properly credited, which ensures maximum dissemination and a wider impact of our publications. The book taken as a whole is c 2018 MDPI, Basel, Switzerland, distributed under the terms and conditions of the Creative Commons license CC BY-NC-ND (http://creativecommons.org/licenses/by-nc-nd/4.0/). Table of Contents About the Special Issue Editor ...................................... v Preface to ”Egg Consumption and Human Health” .......................... vii Jose M. Miranda, Xaquin Anton, Celia Redondo-Valbuena, Paula Roca-Saavedra, Jose A.
    [Show full text]
  • Post-Polymerization Modifications of Polymeric Monolithic Columns: a Review
    Chromatography 2014, 1, 24-53; doi:10.3390/chromatography1010024 OPEN ACCESS chromatography ISSN 2227-9075 www.mdpi.com/journal/chromatography Review Post-Polymerization Modifications of Polymeric Monolithic Columns: A Review Sinéad Currivan and Pavel Jandera * Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, Pardubice 532 10, Czech Republic; E-Mail: [email protected] * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +420-460-037-023. Received: 18 December 2013; in revised form: 16 January 2014 / Accepted: 17 January 2014 / Published: 10 February 2014 Abstract: The vast cache of methods used in polymeric monolithic column modification is presented herein, with specific attention to post-polymerization modification reactions. The modification of polymeric monolithic columns is defined and can include the modification of pre-existing surface groups, the addition of polymeric chains or indeed the addition of structures such as nano-particles and nano-structures. The use of these modifications can result in the specific patterning of monoliths, useful in microfluidic device design or in the investigation of modification optimization. Keywords: polymer monoliths; modifications; HPLC; CEC; grafting; click chemistry; hypercross-linking; nano-particles List of Abbreviations: 2-acrylamido-2-methyl-1-propanesulphonic acid AMPS, α, α’-azoisobutyronitrile AIBN, adenosine triphosphate ATP, atom transfer radical polymerization ATRP, benzoyl peroxide
    [Show full text]
  • Tangential Flow Filtration of Haptoglobin
    Received: 10 January 2020 Revised: 7 April 2020 Accepted: 21 April 2020 DOI: 10.1002/btpr.3010 RESEARCH ARTICLE Tangential flow filtration of haptoglobin Ivan S. Pires | Andre F. Palmer William G. Lowrie Department of Chemical and Biomolecular Engineering, The Ohio State Abstract University, Columbus, Ohio Haptoglobin (Hp) is a plasma glycoprotein that scavenges cell-free hemoglobin (Hb). Correspondence Hp has various potential therapeutic applications, but it has been mainly studied for *Andre F. Palmer, William G. Lowrie treatment of acute hemolytic conditions that can arise from situations such as mas- Department of Chemical and Biomolecular Engineering, The Ohio State University, sive blood transfusion, infusion of stored red blood cells, severe burns, trauma, sepsis, 151 West Woodruff Ave, Columbus, OH radiation injury, and others. Therefore, Hp may also be beneficial during chronic 43210. Email: [email protected] hemolytic disease states such as hereditary spherocytosis, nocturnal hemoglobinuria, sickle-cell anemia, and malaria. Various methods have been developed to purify Hp Funding information National Cancer Institute, Grant/Award from plasma or plasma fractions. However, none of these methods have exploited Number: P30 CA016058; National Heart, the large molecular weight (MW) range distribution of Hp polymers to easily isolate Lung, and Blood Institute, Grant/Award Numbers: R01HL126945, R01HL138116, Hp from other plasma proteins. The present study used tangential flow filtration R56HL123015; National Institute of (TFF) to isolate polymeric Hp from plasma proteins using human Fraction IV (FIV) as Biomedical Imaging and Bioengineering, Grant/ Award Number: R01EB021926; NIH Office of the starting material. After removal of insoluble material from a suspension of FIV the Director, Grant/Award Number: S10 paste, the protein mixture was clarified on a 0.2 μm hollow fiber (HF) TFF filter.
    [Show full text]
  • Life Technologies (India) Pvt. Ltd. 306, Aggarwal City Mall, Opposite M2K Pitampura, Delhi – 110034 (INDIA)
    Product Specification Sheet Chicken conalbumin (ovotransferrin) proteins Antibodies Cat # CECA15-S Rabbit Anti-Chicken conalbumin protein antiserum SIZE: 100 ul Cat # CECA15-C Chicken conalbumin protein control for Western blot SIZE: 100 ul Cat # CECA16-N Chicken conalbumin proteins SIZE: 10 mg liquid at 10 mg/ml in PBS 0.05% azide or in powder form. Allergy to chicken egg or proteins is one of the more frequent Reconstitute powder in PBS at 1-10 mg/ml. It can be used causes of food hypersensitivity in infants and young children. Both positive control for antibody #CECA15-S or for coating IgG and IgA class antibodies may be detected. Ovalbumin ELISA plates. intolerance has been implicated in a number of conditions affecting children. In particular, children with cystic fibrosis show elevated anti-ovalbumin antibodies. Ovalbumin antibodies have For Western blot +ve control (Cat # CECA15-C) is supplied also been noted in some forms of kidney disease. A relationship in SDS-PAGE sample buffer (reduced). Load 10 ul/lane of between food allergy and infantile autism has also been observed. #CECA15-C for good visibility with antibody Cat #CECA15- Children with insulin-dependent diabetes mellitus show an S. Store at –20oC in suitable size aliquots. SDS may enhanced immune response to both β-lactoglobulin and crystallize in cold conditions. It should redissolve by ovalbumin, a phenomenon that may be related to the development warming before taking it from the stock. It should be of the disease. Conditions related to ovalbumin intolerance usually heated once prior to loading on gels. If the product has resolve once egg and egg based foods have been withdrawn from been stored for several weeks, then it may be preferable to the patient's diet.
    [Show full text]
  • Capillary Electrochromatography for Analysis of Proteins and Metalloproteinases
    San Jose State University SJSU ScholarWorks Master's Theses Master's Theses and Graduate Research 2008 Capillary electrochromatography for analysis of proteins and metalloproteinases Vasudha Salgotra San Jose State University Follow this and additional works at: https://scholarworks.sjsu.edu/etd_theses Recommended Citation Salgotra, Vasudha, "Capillary electrochromatography for analysis of proteins and metalloproteinases" (2008). Master's Theses. 3572. DOI: https://doi.org/10.31979/etd.m48v-cr76 https://scholarworks.sjsu.edu/etd_theses/3572 This Thesis is brought to you for free and open access by the Master's Theses and Graduate Research at SJSU ScholarWorks. It has been accepted for inclusion in Master's Theses by an authorized administrator of SJSU ScholarWorks. For more information, please contact [email protected]. CAPILLARY ELECTROCHROMATOGRAPHY FOR ANALYSIS OF PROTEINS AND METALLOPROTEINASES A Thesis Presented to The Faculty of the Department of Chemistry San Jose State University In Partial fulfillment of the Requirements for the Degree Master of Science by Vasudha Salgotra August 2008 UMI Number: 1459696 INFORMATION TO USERS The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleed-through, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. ® UMI UMI Microform 1459696 Copyright 2008 by ProQuest LLC. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code.
    [Show full text]
  • Conalbumin More Resistant to Proteolysis and Ther- the Use Of
    IRON AND PROTEIN KINETICS STUDIED BY MEANS OF DOUBLY LABELED HUMAN CRYSTALLINE TRANSFERRIN * BY JAY H. KATZ (From the Radioisotope and Medical Services of the Boston Veterans Administration Hospital, and the Dept. of Medicine, Boston University School of Medicine, Boston, Mass.) (Submitted for publication June 16, 1961; accepted August 10, 1961) Although proteins isotopically labeled with io- demonstrated that not only is the iron complex of dine have been used in tracer studies for some conalbumin more resistant to proteolysis and ther- years, interest has been mainly focused on their mal denaturation than the metal-free protein, but distribution and degradation in the body. While it that it is possible to maintain the iron-binding ca- is often possible to produce iodoproteins whose pacity of the former after extensive iodination. physical and chemical properties are not grossly Since both iron and transferrin levels are af- altered, the possible effects of iodination on the fected in a variety of disease states, a technique functional properties of such substances in a in which both can be followed simultaneously in physiologic setting have been less thoroughly in- vivo should prove valuable. Elmlinger and co- vestigated. That it is possible to trace label pro- workers (18) have reported in abstract form on teins without significantly reducing their biologi- the use of doubly labeled "iron-binding globulin" cal activities has been demonstrated in the case of in human subjects, but few details were given as insulin (2) and certain anterior pituitary hor- to the methods of preparation and the distribution mones (3, 4). of the two labels.
    [Show full text]
  • Isoelectric Focusing: Sample Pretreatment – Separation – Hyphenation
    Isoelectric Focusing: Sample Pretreatment – Separation – Hyphenation Linda Silvertand ISBN: 978-90-8891-113-2 Printing: www.proefschriftmaken.nl Copyright: ©2009 by Linda Silvertand Cover design: Linda Silvertand - Lak en anthraciet op canvas Niets uit deze uitgave mag verveelvoudigd en/of openbaar gemaakt worden zonder voorafgaande schriftelijke toestemming van de auteur. All rights reserved. No part of this book may be reproduced or transmitted in any form or by any means without written permission of the author and the publisher holding the copyrights of the published articles. Isoelectric Focusing: Sample Pretreatment – Separation – Hyphenation Isoelectrisch Focusseren: Monstervoorbewerking - Scheiding - Koppeling (met een samenvatting in het Nederlands) Proefschrift ter verkrijging van de graad van doctor aan de Universiteit Utrecht op gezag van de rector magnificus, prof. dr. J.C. Stoof, ingevolge het besluit van het college voor promoties in het openbaar te verdedigen op woensdag 23 september 2009 des middags te 2.30 uur door Linda Henriette Hermina Silvertand geboren op 21 juni 1979 te Heerlen Promotor: prof. dr. G.J. de Jong Co-promotor: dr. W.P. van Bennekom This research is part of the IOP Genomics project (STW 06209) “Proteomics on a chip for monitoring auto-immune diseases” and is supported by the Netherlands Research Council for Chemical Sciences (NWO/CW) with financial aid from the Netherlands Technology Foundation (STW). The printing of this thesis was financially supported by: UIPS (Utrecht Institute for Pharmaceutical
    [Show full text]
  • Sites of Formation of the Serum Proteins Transferrin and Hemopexin
    Sites of Formation of the Serum Proteins Transferrin and Hemopexin G. J. Thorbecke, … , K. Cox, U. Muller-Eberhard J Clin Invest. 1973;52(3):725-731. https://doi.org/10.1172/JCI107234. Research Article Sites of synthesis of hemopexin and transferrin were determined by culturing various tissues of rabbits and monkeys in the presence of labeled amino acids. Labeling of the serum proteins was examined by means of autoradiographs of immunoelectrophoretic patterns as well as by precipitation in the test tubes employing immunospecific antisera. Good correlation was seen between the results obtained by the two different methods. The liver was found to be the only site of many tissues studied that synthesized hemopexin. Transferrin production was observed in the liver, submaxillary gland, lactating mammary gland, testis, and ovary. Find the latest version: https://jci.me/107234/pdf Sites of Formation of the Serum Proteins Transferrin and Hemopexin G. J. THORBECKE, H. H. LiEM, S. KNIGHT, K. Cox, and U. MULLER-EBERHARD From the Department of Pathology, New York University School of Medicine, New York 10016, and the Department of Biochemistry, Scripps Clinic and Research Foundation, and Division of Pediatric Hematology, University of California at San Diego, La Jolla, California 92037 A B S T R A C T Sites of synthesis of hemopexin and in iron deficiency and the decrease in inflammatory re- transferrin were determined by culturing various tis- actions (8). sues of rabbits and monkeys in the presence of labeled Preliminary findings (9, 10) suggested that hemo- amino acids. Labeling of the serum proteins was ex- pexin is formed by the liver.
    [Show full text]
  • Power and Limitations of Electrophoretic Separations in Proteomics Strategies
    Power and limitations of electrophoretic separations in proteomics strategies Thierry. Rabilloud 1,2, Ali R.Vaezzadeh 3 , Noelle Potier 4, Cécile Lelong1,5, Emmanuelle Leize-Wagner 4, Mireille Chevallet 1,2 1: CEA, IRTSV, LBBSI, 38054 GRENOBLE, France. 2: CNRS, UMR 5092, Biochimie et Biophysique des Systèmes Intégrés, Grenoble France 3: Biomedical Proteomics Research Group, Central Clinical Chemistry Laboratory, Geneva University Hospitals, Geneva, Switzerland 4: CNRS, UMR 7177. Institut de Chime de Strasbourg, Strasbourg, France 5: Université Joseph Fourier, Grenoble France Correspondence : Thierry Rabilloud, iRTSV/LBBSI, UMR CNRS 5092, CEA-Grenoble, 17 rue des martyrs, F-38054 GRENOBLE CEDEX 9 Tel (33)-4-38-78-32-12 Fax (33)-4-38-78-44-99 e-mail: Thierry.Rabilloud@ cea.fr Abstract: Proteomics can be defined as the large-scale analysis of proteins. Due to the complexity of biological systems, it is required to concatenate various separation techniques prior to mass spectrometry. These techniques, dealing with proteins or peptides, can rely on chromatography or electrophoresis. In this review, the electrophoretic techniques are under scrutiny. Their principles are recalled, and their applications for peptide and protein separations are presented and critically discussed. In addition, the features that are specific to gel electrophoresis and that interplay with mass spectrometry( i.e., protein detection after electrophoresis, and the process leading from a gel piece to a solution of peptides) are also discussed. Keywords: electrophoresis, two-dimensional electrophoresis, isoelectric focusing, immobilized pH gradients, peptides, proteins, proteomics. Table of contents I. Introduction II. The principles at play III. How to use electrophoresis in a proteomics strategy III.A.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 8,940,687 B2 Naidu Et Al
    USOO8940687B2 (12) United States Patent (10) Patent No.: US 8,940,687 B2 Naidu et al. (45) Date of Patent: Jan. 27, 2015 (54) ANTIMICROBIAL ANGIOGENIN 5,286,487 A 2/1994 Vallee et al. COMPLEXES (ANGEX) AND USES THEREOF 7,074,7596,172,040 B2B1 7,1/2001 2006 Naidu 7,125,963 B2 10/2006 Naidu (71) Applicant: Naidu LP, Pomona, CA (US) 7,601,689 B2 10/2009 Naidu 7.956,031 B2 6, 2011 Naidu et al. (72) Inventors: A. Satyanarayan Naidu, Diamond Bar, 8,003,603 B2 8/2011 Naidu CA (US); A. G. Tezus Naidu, Santa 8,021,659 B2 9, 2011 Naidu et al. Ana, CA (US); A. G. Sreus Naidu, 8,309,080 B2 11/2012 Naidu et al. Santa Ana, CA (US 2007, 0141101 A1 6/2007 Nugent et al. anta Ana, CA (US) 2008/02540.18 A1 10, 2008 Naidu 2011/0200575 A1 8, 2011 Naidu et al. (73) Assignee: Naidu LP, Pomona, CA (US) 2011/0286986 A1 11/2011 Naidu et al. (*) Notice: Subject to any disclaimer, the term of this OTHER PUBLICATIONS patent is extended or adjusted under 35 U.S.C. 154(b) by 0 days. González-Chávez et al. ((International Journal of Antimicrobial 21) Appl. No.: 13/861,236 Agents 33 (2009) 301.e1-301.e8).* (21) ppl. No.: 9 Acharya, et al. “Crystal Structure of Human Angiogenin Reveals the Structural Basis for its Functional Divergence from Ribonuclease.” (22) Filed: Apr. 11, 2013 Proc. Natl. Acad. Sci. USA, vol. 91, pp. 2915-2919, Apr. 1994. O O Hu, et al.
    [Show full text]
  • High Performance Capillary Electrophoresis
    High Performance Capillary Electrophoresis A Primer A Primer High Performance Capillary Electrophoresis Second completely revised edition by Henk H. Lauer and Gerard P. Rozing Foreword Electrophoresis was born at the beginning of nineteenth century, even earlier than chromatography. It has taken a long development path, which in the early days was framed by names like Kohlrausch, Tiselius and later Everaerts. At the beginning of eighties in the last century, Jorgenson cause a paradigm shift by executing electrophoresis in a very thin capillary with inner diameter less then 100 μm leading to capillary electrophoresis. Since then, capillary electrophoresis has been constantly improved and has become a routine technique. Instrument manufacturers like Agilent Technologies have developed instrumentation for capillary electrophoresis serving many analytical laboratories reliably for many years. After a decline at the beginning of this decade, the capillary electrophoresis market is recovering strongly. Especially, hyphenation with mass spectrometers has been very fruitful and brought a remarkable ability to unravel complex structures of biomolecules with invincible speed and sensitivity. And there are several modes of capillary electrophoresis, which make it useful for separation of really wide class of compounds, such as small ions, peptides, proteins, DNA fragments both sequencing and restriction ones, and even complete cells or uncharged molecules. When compared with liquid chromatography capillary electrophoresis has one big advantage: the separation process takes place in a rather simple environment – in homogeneous solutions or in a polymeric sieving network and in a separation space with simple geometry, like a cylindrical channel. The equations describing completely movement of matter in space and time can be than easily formulated and solved by advanced mathematical methods.
    [Show full text]