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Protein Characterization By Capillary Isoelectric Focusing Electrophoresis Reversed Phase Liquid Chromatography And Mass Spectrometry
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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 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 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 New Methods in Peptide Mapping for the Characterization of Proteins by : William S. Hancock
Download or read book New Methods in Peptide Mapping for the Characterization of Proteins written by William S. Hancock and published by CRC Press. This book was released on 1995-10-23 with total page 280 pages. Available in PDF, EPUB and Kindle. Book excerpt: This text is devoted to the characterization of recombinant DNA-derived proteins by peptide mapping. It describes new technological procedures including capillary electrophoresis, analysis of glycopeptides and the use of electrospray and matrix-assisted laser desorption mass spectrometry. The book presents practical procedures for preparing a protein sample, the enzyme digestion, choice of separation method and procedures for the structural analysis of the separated species. Many figures of peptide maps illustrate typical results. Tables of summary information about digestion, separation conditions, and analyses of important protein samples are also presented.
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 High Resolution Separation and Analysis of Biological Macromolecules by : Barry L. Karger
Download or read book High Resolution Separation and Analysis of Biological Macromolecules written by Barry L. Karger and published by Elsevier. This book was released on 1996 with total page 664 pages. Available in PDF, EPUB and Kindle. Book excerpt: The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. More than 260 volumes have been published (all of them still in print) and much of the material is relevant even today--truly an essential publication for researchers in all fields of life sciences. Key Features * Liquid chromatography * Electrophoresis * Mass spectrometry.
Book Synopsis Capillary Electrophoresis of Proteins by : Tim Wehr
Download or read book Capillary Electrophoresis of Proteins written by Tim Wehr and published by CRC Press. This book was released on 1998-10-20 with total page 301 pages. Available in PDF, EPUB and Kindle. Book excerpt: "Provides practical information on the application of capillary electrophoresis (CE) to protein analysis, with an emphasis on developing and optimizing CE techniques in the laboratory. Includes separation methods bases on mass, charge, isoelectric point, molecular sieving, and affinity interactions."
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 Novel Micro-separation Techniques and Their Couplings to Mass Spectrometry for the Analysis of Complex Peptides and Proteins by :
Download or read book Novel Micro-separation Techniques and Their Couplings to Mass Spectrometry for the Analysis of Complex Peptides and Proteins written by and published by . This book was released on 2013 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Mass spectrometry (MS)-based technology has evolved as a powerful tool for the analysis of a wide array of biomolecules. However, its application to complex peptidomics and proteomics samples, for example neuropeptides, has proven to be challenging due to their extreme chemical complexities and low abundances in biological tissue samples. This PhD dissertation is focused on addressing some of these challenges by developing novel micro-separation tools and coupling them with MS to achieve enhanced MS sensitivity and increased peptide and protein coverage with lower sample consumption and reduced experimental time. Both chromatography-based and electrophoresis-based separations as well as their couplings to MS have been explored in this work, including monolithic material-based chromatography, improved capillary electrophoresis (CE) separation and novel coupling of separations to mass spectrometric imaging. N-acryloxysuccinimide-based monolithic column has been fabricated with specific antibodies immobilized onto the column to enable the extraction of targeted neuropeptide family from biological matrices. Alternatively, enzymes were immobilized onto the column for highly efficient on-column enzymatic digestion for proteomics studies. A methacrylate-based monolithic column with C12 functional groups was developed and coupled to both electrospray ionization (ESI) MS and matrix-assisted laser desorption/ionization (MALDI) MS for reversed-phase liquid chromatography (RPLC). For capillary electrophoresis, two improved capillary isoelectric focusing (CIEF) systems have been developed, including membrane-assisted CIEF and immobilized pH-gradient CIEF to obtain reduced sample loss and enhanced MS signal for neuropeptide analysis. An improved sheathless pressure-assisted CE (PACE) system was also developed for coupling to MALDI MS. For novel separation-MS imaging coupling, three generations of interfaces have been designed to couple CE, LC and multi-dimensional separations to MS imaging (MSI) for quantitative analysis of complex peptides. Collectively, this work results in development of the new generation of micro-separation and sample preparation tools and their coupling to various mass spectrometric ionization and detection modes for sensitive and rapid analysis of complex peptides and proteins. The technological innovations have significantly expanded the toolbox for peptidomics/proteomics analysis and will enable more in-depth investigations on peptide function and regulation.
Book Synopsis Development of Multidimensional Liquid Phase Protein Separations for Proteomic Analysis by : Bobby Fachrizal Assiddiq
Download or read book Development of Multidimensional Liquid Phase Protein Separations for Proteomic Analysis written by Bobby Fachrizal Assiddiq and published by . This book was released on 2009 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Book Synopsis Isoelectric Trapping and Mass Spectrometry by : Stephanie Marie Cologna
Download or read book Isoelectric Trapping and Mass Spectrometry written by Stephanie Marie Cologna and published by . This book was released on 2012 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Mass spectrometry (MS) has played a major role in the proteomic analysis of an array of biological samples. Even so, inherent limitations exist such as sample complexity and the dynamic range. In an attempt to overcome these limitations, prefractionation is typically performed followed by reversed phase liquid chromatography coupled with MS. Pre-fractionation can be performed in several formats including chromatographic or electrophoretic based methods. Solution-based isoelectric point (pI) fractionation, specifically isoelectric trapping (IET), provides an attractive alternative for pre-fractionation in bottom-up proteomic studies. A recently developed device, membrane separated wells for isoelectric focusing and trapping (MSWIFT), provides rapid separation on the basis of pI and resulting solutions are MS compatible without the need for extensive sample cleanup. Initial experiments demonstrate fractionation using MSWIFT, of peptide mixtures ranging from standards to a yeast lysate where resulting fractions are analyzed using matrixassisted laser desorption/ionization (MALDI) -- MS or further separated using reversed phase liquid chromatography followed by tandem MS (MS/MS) analysis. Identified yeast proteins range in size, pI and copy number illustrating an ability to increase the depth of proteome coverage when using MSWIFT. Extensive studies were also performed using MSWIFT in a multi-stage fractionation platform to improve peptide and protein identifications for the first large-scale proteomic study of the model fungus, Neurospora crassa. A second focus of this work is the development of a new sample preparation method for proteolytic digestion and high-throughput separations using MSWIFT. Histidine is used as a neutral pH, isoelectric, sample buffer for tryptic digestion of proteins and also assists in rapid separations using MSWIFT owing to the low conductivity. Tryptic digests of individual standard proteins and a mixture of standard proteins are used to illustrate these advantages. Finally, the histidine buffer sample preparation method is incorporated into a two-dimensional separation strategy. Tryptic peptides are fractionated using MSWIFT and resulting solutions are further separated using capillary electrophoresis (CE) coupled with MALDI-MS/MS. Performing the two-dimensional strategy allows for increased confidence in peptide and protein assignment owing to experimentally determined in-solution charge states and estimated pI values.
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 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 Humana. This book was released on 2010-11-10 with total page 0 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 Protein Structure Analysis by : Roza Maria Kamp
Download or read book Protein Structure Analysis written by Roza Maria Kamp and published by Springer Science & Business Media. This book was released on 2012-12-06 with total page 311 pages. Available in PDF, EPUB and Kindle. Book excerpt: "Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.
Book Synopsis Multidimensional Liquid Chromatography by : Steven A. Cohen
Download or read book Multidimensional Liquid Chromatography written by Steven A. Cohen and published by John Wiley & Sons. This book was released on 2008-05-09 with total page 490 pages. Available in PDF, EPUB and Kindle. Book excerpt: Multidimensional Liquid Chromatography (MDLC) is a very powerful separation technique for analyzing exceptionally complex samples in one step. This authoritative reference presents a number of recent contributions that help define the current art and science of MDLC. Topics covered include instrumentation, theory, methods development, and applications of MDLC in the life sciences and in industrial chemistry. With the information to help you perform very difficult separations of complex samples, this reference includes chapters contributed by leading experts or teams of experts.
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 2018-06-28 with total page 74 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 Analysis and Identification of Proteins from Human Cells Using Mass Spectrometry Combined with Liquid Chromatography, Electrophoresis, and Database Searching by : Yajuan Chen
Download or read book Analysis and Identification of Proteins from Human Cells Using Mass Spectrometry Combined with Liquid Chromatography, Electrophoresis, and Database Searching written by Yajuan Chen and published by . This book was released on 1999 with total page 302 pages. Available in PDF, EPUB and Kindle. Book excerpt:
