Capillary Electrophoresis Of Proteins And Peptides

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.

"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."

The goal of this book is to show recent developments in the CE analysis of protein pharmaceuticals. It is devoted completely to practical concerns to strengthen the use of CE within the biotechnology industry, highlighting the uses of CE in various areas of product development including formulation studies, process development, product characterization and validated lot release and stability testing.

This text aims to evaluate the actual impact of high-performance capillary electrophoresis on analytical biotechnology and environmental analysis. The first part of the book presents a survey of present innovations in instrument design and different methods of pre-concentration techniques in order to obtain increased separations at higher sensitivities. The second part contains articles on applications of HPCE to protein and peptide analysis. In the third part, applications of HPCE in the investigation of drug abuse and drug interactions are presented. The last two parts of the book deal with the use of HPCE at low-UV wavelengths and negative-UV absorption. The book should be of interest to those working in HPCE research and applications.

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.

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.