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Capillary Isoelectric Focusing Electrospray Ionization Mass Spectrometry For Protein Characterization
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Book Synopsis Capillary Isoelectric Focusing-electrospray Ionization Mass Spectrometry for Protein Characterization by : Qing Tang
Download or read book Capillary Isoelectric Focusing-electrospray Ionization Mass Spectrometry for Protein Characterization written by Qing Tang and published by . This book was released on 1996 with total page 246 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Combination of Capillary Isoelectric Focusing and Liquid Chromatography with Electrospray Ionization Mass Spectrometry for Protein Characterization by : Jing Wei
Download or read book Combination of Capillary Isoelectric Focusing and Liquid Chromatography with Electrospray Ionization Mass Spectrometry for Protein Characterization written by Jing Wei and published by . This book was released on 1998 with total page 228 pages. Available in PDF, EPUB and Kindle. Book excerpt: Combination of capillary isoelectric focusing (CIEF) and liquid chromatography (LC) with electrospray ionization mass spectrometry (ESIMS) is utilized for protein characterization. Both CIEF-ESIMS and LC-MS are studied for phosphoprotein characterization. At the intactprotein level, on-line CIEF-ESIMS is applied for phosphorylation analysis to obtain the two dimensional information: isoelectric point and molecular mass. At the peptide level, on-line LC-ESI-MS and LC-ESI neutral loss MS/MS were highly specific for the determination of sites of phosphorylation and successfully applied on the tryptic digest of tau proteins phosphorylated by different kinases. On-line combination of CIEF with ESI-TOF MS is demonstrated for high resolution analysis of some model proteins and human hemoglobin variants. The successful coupling of these two techniques optimizes the efficiency of CIEF separation since TOF MS is able to produce a full spectrum in the shortest response time among all kinds of mass analyzers.
Book Synopsis Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry by : Feng Zhou
Download or read book Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry written by Feng Zhou and published by ProQuest. This book was released on 2008 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: With the development of genomics, separation technology and mass spectrometry, proteomics has become an important tool in biological research. Characterization of posttranslational modification (PTM) by mass spectrometry is attracting an increasing attention because of its importance in signal transduction and the difficulty in obtaining this information from genomic methods. Bottom-up methods, which analyze the peptides from enzymatic digestion of proteins, are widely used for proteomic research. However, bottom-up methods may lose some important information when characterizing PTM since not all peptides are detected. In contrast, top-down methods directly characterize intact proteins, which makes it possible to retain PTM information. This dissertation focuses on the characterization of proteins by coupling high throughput methods, capillary isoelectric focusing electrophoresis (CIEF) and electrospray ionization (ESI) mass spectrometry (MS). In this work, a CIEF-RPLC-ESIMS system is designed and constructed for 2-D profiling of protein mixture. The microdialysis membrane based cathodic cell used in this system not only serves as the cathodic cell in CIEF separation, but also transfers the focused proteins separated by CIEF to RPLC. After eliminating the interference of ampholyte, a few fmol of protein can be detected with this system. The detection limit of this system is one order of magnitude lower than 2-D PAGE. By taking the advantage of high resolving power with mass spectrometry and multidimensional separation, the theoretical peak capacity of this system can reach 1x10 6, which is two orders of magnitude higher than theoretical peak capacity of 2-D PAGE. This system is firstly tested with a standard protein mixture. After slight modification, this system is used to profile the intact protein from a yeast enzyme concentrate and a lysate of the green sulfur bacterium Chlorobium tepidum. The pI vs. MW profile obtained from CIEF-RPLC-MS compares favorably with theoretical data derived from C. tepidum genome and experimental data obtained from 2-D-PAGE.
Book Synopsis On-line Coupling of Capillary Electrophoresis and Liquid Chromatography with Electrospray Ionization Mass Spectrometry for Protein Characterization by : Liyu Yang
Download or read book On-line Coupling of Capillary Electrophoresis and Liquid Chromatography with Electrospray Ionization Mass Spectrometry for Protein Characterization written by Liyu Yang and published by . This book was released on 1998 with total page 218 pages. Available in PDF, EPUB and Kindle. Book excerpt: On-line coupling of capillary isoelectric focusing (CIEF) and liquid chromatography (LC) with electrospray ionization mass spectrometry (ESIMS) is utilized for protein characterization. A microdialysis junction, based on a microdialysis membrane connecting both a separation capillary and a short, sharply tapered microelectrospray emitter capillary, is demonstrated for CIEF-ESIMS. Besides the electrical connection across the microdialysis junction, post-run acidification via the dialysis membrane is demonstrated for enhancing the protonation and the ionization efficiency of focused proteins while maintaining separation efficiency, resolution, and sensitivity in CIEF-ESIMS.
Book Synopsis Capillary Electrophoresis of Proteins and Peptides by : Mark A. Strege
Download or read book Capillary Electrophoresis of Proteins and Peptides written by Mark A. Strege and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 340 pages. Available in PDF, EPUB and Kindle. Book excerpt: Throughout the more than 20 years that have followed the beginnings of capillary electrophoresis (CE), its application to the analysis of proteins and peptides has continued to be reliable, versatile, and productive. Over time, CE has matured to become a superb complement to HLPC, and in many cases has also evolved as an automated and quantitative replacement for conventional slab gel electrophoresis methods such as SDS-PAGE and isoelectric focusing. Within Capillary Electrophoresis of Proteins and Peptides, we have assembled contributions from researchers who are applying state-of-the-art CE for protein and peptide analysis, including topics that we believe are of great potential both in the present and for the future. In comparison to traditional separation methods, CE represents a miniaturized analysis technique (especially in its microchip-based format) that is highly dependent upon the basic fundamentals of effective sample recovery and high sensitivity detection. With these issues in mind, Chapters 1–4 describe recently developed approaches for both capillary coatings and analyte detection via laser-induced fluorescence. Since the discipline of biotechnology has established itself as a primary platform for the application of CE to the analysis of proteins and peptides, Chapters 5–7 demonstrate a variety of examples of the specific techniques that have been applied for the development of biopharmaceuticals and their commercialization. The methods covered here include also the analysis of oligosaccharides from glycoproteins.
Book Synopsis State-of-art Separation Mass Spectrometry in Protein Characterization by : Siyuan Liu
Download or read book State-of-art Separation Mass Spectrometry in Protein Characterization written by Siyuan Liu and published by . This book was released on 2016 with total page 223 pages. Available in PDF, EPUB and Kindle. Book excerpt: A therapeutic protein derived from recombinant DNA technology usually poses a high degree of complexity. A complete heterogeneity profile of the therapeutic protein is critical to ensure the quality, safety and efficacy of the drug. To achieve a complete mapping of all possible protein modifications including the various types of post-translational modifications and sequence variants, the incorporation of high resolution separation methods and high sensitivity detection techniques are required. The purpose of this dissertation is to develop and apply sensitive and robust mass spectrometry (MS)-based analytical platforms to address the challenges associated with protein identification and characterization. In the first chapter, the current analytical techniques, including various gel-based, chromatographic-based and capillary electrophoresis-based separation methods frequently incorporated with mass spectrometry for protein identification and characterization are reviewed. The traditional sample preparation workflows for MS-based protein identification are introduced. In addition, the application of mass spectrometry for the determination of protein post-translational modifications and the associated analytical challenges are discussed. The sample preparation steps involved in the LC-MS analysis of a therapeutic protein can potentially introduce disulfide scrambling leading to the incorrect characterization of the disulfide linkage structure. Chapter 2 focuses on the development of an LC-MS work flow for the determination of the disulfide mapping of proteins that are sensitive to disulfide scrambling. The recombinant human neublastin, a potential therapeutic agent with disulfide linkages partially elucidated via the traditional non-reduced multi-enzyme digestion strategy is studied. The modified workflow includes the optimization of the partial reduction and alkylation steps prior to multi-enzyme digestion in order to minimize the breakage of a relatively unstable intermolecular disulfide bond and the rapid thiol-disulfide exchange between adjacent cysteines. Using the developed work flow, while scrambling remained, complete and reproducible disulfide mapping was achieved via in-depth LC-MS analysis with CID and ETD fragmentations. In Chapter 3, a powerful methodology combining metabolic labeling, SDS-PAGE protein fractionation and nano LC-MS analysis is applied to isolate and identify the potential protein target to a novel small molecule drug from cell lysates. The non-covalent binding between the identified protein target and the drug is demonstrated with comprehensive characterization of the potential target. In Chapter 4, a new design of a low-sheath-flow liquid junction interface is described to integrate commercial capillary electrophoretic instrumentation and electrospray ionization mass spectrometer for automatic peptide and protein analysis. Different separation modes including capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF) were applied to couple with mass spectrometry via the liquid junction interface. Confident identification and high separation efficiency of a BSA digest are demonstrated with the integrated CZE-MS system. In addition, separation of a mixture of model proteins with closely related isoelectric points was achieved with CIEF-MS analysis leading to the identification of different types of post-translational modifications. The system was also employed to study the potential degradation products of a therapeutic protein, recombinant human growth hormone. The low-sheath-flow liquid junction interface integrated CIEF-ESI-MS system potentially can be an alternative technique to the well-established LC-MS method for the online enrichment and identification of the charge heterogeneity in biopharmaceuticals. Chapter 5 summarizes the work in the dissertation and provides a brief discussion on the future direction of each project.
Book Synopsis Capillary Electrophoresis-Mass Spectrometry by : James Q. Xia
Download or read book Capillary Electrophoresis-Mass Spectrometry written by James Q. Xia and published by Springer. This book was released on 2016-11-08 with total page 80 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book covers the latest developments in capillary electrophoresis-mass spectrometry for the analysis of therapeutic proteins. The application of capillary electrophoresis-mass spectrometry (CE-MS) coupling technology in the analysis of recombinant therapeutic proteins is detailed thoroughly. Specific topics include recent developments in coupling capillary electrophoresis with mass spectrometry for the quality control of monoclonal antibody therapeutics, top-down analysis of monoclonal antibody using the CE-MS platform, and detection of host cell protein impurities. Comprehensive characterization of antibody-drug conjugates (ADCs) by coupling capillary electrophoresis with mass spectrometry is also covered. This is an ideal book for scientists in the life science and biopharmaceutical industry who are working on characterizing the PTMs of monoclonal antibodies, as well as graduate students and researchers in the separation science and biological mass spectrometry fields.
Book Synopsis Capillary Gel Electrophoresis by : Andras Guttman
Download or read book Capillary Gel Electrophoresis written by Andras Guttman and published by Newnes. This book was released on 2021-12-04 with total page 391 pages. Available in PDF, EPUB and Kindle. Book excerpt: Capillary Gel Electrophoresis and Related Microseparation Techniques covers all theoretical and practical aspects of capillary gel electrophoresis. It also provides an excellent overview of the key application areas of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods. It not only gives readers a better understanding of how to utilize this technology, but also provides insights into how to determine which method will provide the best technical solutions to particular problems. This book can also serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses. Covers all theoretical and practical aspects of capillary gel electrophoresis Excellent overview of the key applications of nucleic acid, protein and complex carbohydrate analysis, affinity-based methodologies, micropreparative aspects and related microseparation methods Teaches readers how to use the technology and select methods that are ideal for fundamental problems Can serve as a textbook for undergraduate and graduate courses in analytical chemistry, analytical biochemistry, molecular biology and biotechnology courses
Book Synopsis Microcharacterization of Proteins by : Roland Kellner
Download or read book Microcharacterization of Proteins written by Roland Kellner and published by John Wiley & Sons. This book was released on 2008-09-26 with total page 346 pages. Available in PDF, EPUB and Kindle. Book excerpt: Proteomics - the analysis of the whole set of proteins and their functions in a cell - is based on the revolutionary developments which have been achieved in protein analysis during the last years. The number of finished genome projects is growing and in parallel there is a dramatically increasing need to identify the products of revealed genes. Acting on a micro level modern protein chemistry increases our understanding of biological events by elucidating the relevant structure-function relationships. The second edition of the successful title Microcharacterization of Proteins presents a current overview of modern protein analysis: From sample preparation to sequence analysis, mass spectrometry and bioinformatics it informs about the tools needed in protein research. This makes the book indispensable for everyone involved in proteomics!
Book Synopsis Development of Capillary Electrophoresis and Mass Spectrometry Methods in Protein Analysis by : Lin Liu
Download or read book Development of Capillary Electrophoresis and Mass Spectrometry Methods in Protein Analysis written by Lin Liu and published by . This book was released on 1999 with total page 242 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Applied Electrospray Mass Spectrometry by : Birendra N. Pramanik
Download or read book Applied Electrospray Mass Spectrometry written by Birendra N. Pramanik and published by CRC Press. This book was released on 2002-02-28 with total page 478 pages. Available in PDF, EPUB and Kindle. Book excerpt: Discussing strategies to determine the structure and machanisms of numerous compound classics, this book covers new chemical and elctrophoretic techniques for rapid sample preconcentration and separation. It summarizes breakthroughs in the theory and instrumentation of electrospray mass spectrometry in pharmaceutical and biomedical applications, provides practical examples for the characterization of peptides, proteins, and glycoproteins, includes applications in proteomics, combinatorial chemistry, and drug characterization. Topics include chemical and electrophoretic techniques for rapid sample preconcentration and separation, screening processes for proteins from libraries of compounds, protein folding and dynamics, and more.
Book Synopsis Mass Spectrometry Analysis for Protein-Protein Interactions and Dynamics by : M. Chance
Download or read book Mass Spectrometry Analysis for Protein-Protein Interactions and Dynamics written by M. Chance and published by John Wiley & Sons. This book was released on 2008-09-22 with total page 325 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presents a wide variety of mass spectrometry methods used to explore structural mechanisms, protein dynamics and interactions between proteins. Preliminary chapters cover mass spectrometry methods for examining proteins and are then followed by chapters devoted to presenting very practical, how-to methods in a detailed way. Includes footprinting and plistex specifically, setting this book apart from the competition.
Book Synopsis Characterization of Protein Therapeutics using Mass Spectrometry by : Guodong Chen
Download or read book Characterization of Protein Therapeutics using Mass Spectrometry written by Guodong Chen and published by Springer Science & Business Media. This book was released on 2014-07-08 with total page 408 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book highlights current approaches and future trends in the use of mass spectrometry to characterize protein therapies. As one of the most frequently utilized analytical techniques in pharmaceutical research and development, mass spectrometry has been widely used in the characterization of protein therapeutics due to its analytical sensitivity, selectivity, and specificity. This book begins with an overview of mass spectrometry techniques as related to the analysis of protein therapeutics, structural identification strategies, quantitative approaches, followed by studies involving characterization of process related protein drug impurities/degradants, metabolites, higher order structures of protein therapeutics. Both general practitioners in pharmaceutical research and specialists in analytical sciences will benefit from this book that details step-by-step approaches and new strategies to solve challenging problems related to protein therapeutics research and development.
Book Synopsis Proteomics and Peptidomics by : Gyorgy Marko-Varga
Download or read book Proteomics and Peptidomics written by Gyorgy Marko-Varga and published by Elsevier. This book was released on 2005-12-23 with total page 663 pages. Available in PDF, EPUB and Kindle. Book excerpt: Proteomics and peptidomics is the detailed understanding of the role that proteins and peptides play in health and disease and is a necessary compliment to genetic analysis. The functional expression analysis of both proteins and peptides plays a central role in modern drug discovery as well as drug development, and is also a key research area in systems biology. Proteomics and Peptidomics captures the width as well as the depth within the area and exemplifies the variety as well as the traditional basis of analytical chemistry that is needed in order to move forward in expression analysis studies. As a fast emerging field, it gives and overview of parts within the field combined with highly specialized and dedicated topics that are intended to compliment each other.
Book Synopsis Protein Analysis by Capillary Isolectric Focusing Combined with Mass Spectrometry by : Anne Chartogne
Download or read book Protein Analysis by Capillary Isolectric Focusing Combined with Mass Spectrometry written by Anne Chartogne and published by . This book was released on 2002 with total page 167 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Electrifying the Molecules of Life by : Magnus Wetterhall
Download or read book Electrifying the Molecules of Life written by Magnus Wetterhall and published by . This book was released on 2004 with total page 44 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Protein Analysis using Mass Spectrometry by : Mike S. Lee
Download or read book Protein Analysis using Mass Spectrometry written by Mike S. Lee and published by John Wiley & Sons. This book was released on 2017-05-26 with total page 282 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presents Practical Applications of Mass Spectrometry for Protein Analysis and Covers Their Impact on Accelerating Drug Discovery and Development Covers both qualitative and quantitative aspects of Mass Spectrometry protein analysis in drug discovery Principles, Instrumentation, Technologies topics include MS of peptides, proteins, and ADCs , instrumentation in protein analysis, nanospray technology in MS protein analysis, and automation in MS protein analysis Details emerging areas from drug monitoring to patient care such as Identification and validation of biomarkers for cancer, targeted MS approaches for biomarker validation, biomarker discovery, and regulatory perspectives Brings together the most current advances in the mass spectrometry technology and related method in protein analysis
