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The characterization of peptides and proteins is central to understanding their function and expression in biological matrices. Moreover, these macromolecules are important biomarkers of many human diseases. In recent years, the performance of separation techniques based on electromigration have significantly increased. The development of microdevices has reduced sample consumption and waste production while high-sensitivity detectors, such as mass spectrometry (MS) or laser-induced fluorescence (LIF), have significantly improved with regards to separation efficiency and detection limits. All of these advancements have led to appreciably enlarged fields of application. Nowadays, a multitude of studies using separation techniques based on electromigration to study proteins and peptides from numerous real matrices are available in the literature. This Special Issue covers the most recent knowledge and advances in the study of peptides and proteins using several electrophoresis techniques, as well as the characterization of relevant proteins and peptides in application areas such as clinical studies, functional foods, and toxicology.
The characterization of peptides and proteins is central to understanding their function and expression in biological matrices. Moreover, these macromolecules are important biomarkers of many human diseases. In recent years, the performance of separation techniques based on electromigration have significantly increased. The development of microdevices has reduced sample consumption and waste production while high-sensitivity detectors, such as mass spectrometry (MS) or laser-induced fluorescence (LIF), have significantly improved with regards to separation efficiency and detection limits. All of these advancements have led to appreciably enlarged fields of application. Nowadays, a multitude of studies using separation techniques based on electromigration to study proteins and peptides from numerous real matrices are available in the literature. This Special Issue covers the most recent knowledge and advances in the study of peptides and proteins using several electrophoresis techniques, as well as the characterization of relevant proteins and peptides in application areas such as clinical studies, functional foods, and toxicology.
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.
This book provides a comprehensive survey of recent developments and applications of high performance capillary electrophoresis in the field of protein and peptide analysis with a distinct focus on the analysis of intact proteins. With practical detail, the contents cover different modes of capillary electrophoresis (CE) useful for protein and peptide analysis, CZE, CIEF, ACE, CGE, and different types of application such as the quality control of therapeutic proteins and monoclonal antibodies, clinical analyses of chemokines in tissues, qualitative and quantitative analysis of vaccine proteins, and determination of binding constants in complexes involving peptides or proteins. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and exhaustive, Capillary Electrophoresis of Proteins and Peptides: Methods and Protocols serves both beginners and experts with a collection of the current and most active topics in this vital field of study.
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.
Furthering efforts to simulate the potency and specificity exhibited by peptides and proteins in healthy cells, this remarkable reference supplies pharmaceutical scientists with a wealth of techniques for tapping the enormous therapeutic potential of these molecules-providing a solid basis of knowledge for new drug design. Provides a broad, comprehensive overview of peptides and proteins as mediators of cell movement, proliferation, differentiation, and communication. Written by more than 50 leading international authorities, Peptides and Protein Drug Analysis discusses strategies for dealing with the complexity of peptides and proteins in conformational flexibility and amino acid sequence variability analyzes drug formulations facilitated by solid-phase peptide synthesis and recombinant DNA technology examines chemical purity analysis by high-pressure chromatographic, capillary electrophoretic, gel electrophoretic, and isoelectric focusing methods highlights drug design elements derived from protein folding, bioinformatics, and computational chemistry demonstrates uses of unnatural mutagenesis and combinatorial chemistry explores mass spectrometry, protein sequence, and carbohydrate analysis illustrates bioassays and other new functional analysis methods surveys spectroscopic techniques such as ultraviolet, fluorescence, Fourier transform infrared, and nuclear magnetic resonance (NMR) addresses ways of distinguishing between levels of therapeutic and endogenous agents in cells reviews structural analysis tools such as ultracentrifugation and light, X-ray, and neutron scattering and more! Featuring over 3400 bibliographic citations and more than 500 tables, equations, and illustrations, Peptide and Protein Drug Analysis is a must-read resource for pharmacists; pharmacologists; analytical, organic, and pharmaceutical chemists; cell and molecular biologists; biochemists; and upper-level undergraduate and graduate students in these disciplines.
Abstract: A reference text for food scientists, analysts, and biochemists presents a systematic, comprehensive survey of working procedures and methods for the analysis of proteins, peptides, and amino acids of varying origin and composition that can be used with equipment that generally is available in the average laboratory. Topics include: general laboratory methods for the preparation and characterization of proteinaceous materials; electrophoreticmethods (especially using polyacrylamide gels); thin-layer chromatographic methods; gel chromatography and gel filtration methods; and experimental procedures for nitrogendetermination, amino acid composition, electrophoresis, isoelectric focusing, ion-exchange chromatography, and gel filtration. Step-by-step descriptions are provided for sample preparation for analysis and problems requiring resolution are discussed.
Why a Second Edition? The Second Edition provides practical answers to the general question, "How can I obtain useful sequence information from my protein or peptide?" rather than the more specific question asked in the first edition, "How can I obtain the N-terminal sequence?" Important new methods include ways of dealing with blocked N termini, computer analysis of protein sequences, and the recent revolution in mass spectrometry. Mass spectrophotometric characterization of proteins and peptides N-terminal sequencing of proteins with blocked N termini Internal amino acid sequence analysis after protease digestion in-gel and on-blot Improved microscale peptide purification methods Computer analysis of protein sequences New protocols tested and refined through everyday use in authors' laboratories Updated reference chapter covering all aspects of protein microsequencing
This new edition of Gel Electrophoresis of Proteins is a completely new text, with eight of the ten chapters written by new authors. It presents the best methods, hints and tips for core procedures such as one- dimensional polyacrylamide gel electrophoresis, isoelectric focusing, two-dimensional gel electrophoresis, preparative gel electrophoresis, and peptide mapping, complete with the latest refinements and updates of the procedures. In addition, it describes major new techniques which have come to the fore since the previous edition. Thus there are chapters on capillary gel electrophoresis, sequence analysis of gel-resolved proteins, fluorophore-labelled saccharide electrophoresis, and analysis of protein:protein interactions by gel electrophoresis. One thing has not changed. The emphasis is still on describing the best methods, in step-by-step detail, with copious advice to ensure that each method works first time in the reader's hands. The first two editions of Gel Electrophoresis of Proteins: A Practical Approach each gained a strong reputation as easy-to-follow laboratory manuals written by experienced researchers for researchers. The methods were presented in a clear accessible format and had been fully tested to ensure success in the lab. This new edition will strengthen the reputation of the book still further. It is a 'must have' for all those who currently use gel electrophoresis or who plan to do so.
"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."