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 14 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 57 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.

Includes supplementary material: sn.pub/extras

Klappentext
Capillary electrophoresis (CE) is one of the premier methods for the rapid analysis of pharmaceutical proteins and peptides, and thus now even more important with the sequencing of the human genome and the subsequent advent of proteomics. In Capillary Electrophoresis of Proteins and Peptides, leading academic scientists and expert laboratorians working in the pharmaceutical industry describe their best CE techniques in step-by-step detail for easy reproduction. The authors present cutting-edge techniques for both capillary coatings and analytical detection via laser-induced fluorescence, for the development and commercialization of biopharmaceuticals, and for affinity capillary electrophoresis in the evaluation of protein binding, including the use of protein charge ladders. Additional chapters discuss CE and capillary isoelectric focusing, combined with electrospray mass spectrometry detection, for performing proteomic studies. Areas of special interest covered include proteomic applications, as well as applications in the field of protein-ligand binding. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and state-of-the-art, Capillary Electrophoresis of Proteins and Peptides will prove essential to both new and experienced investigators carrying out significant protein and peptide studies in biotechnology, drug discovery, and pharmaceutical research and development.

Inhalt
Surfactant-Based Methods for Prevention of Protein Adsorption in Capillary Electrophoresis.- Capillary Coating for Protein Separation Based on Si-O and Si-C Covalent Bond Formation for Capillary Electrophoresis With Laser-Induced Fluorescence Detection.- On-Column Labeling Reaction for Analysis of Protein Contents of a Single Cell Using Capillary Electrophoresis With Laser-Induced Fluorescence Detection.- Covalent and Noncovalent Labeling Schemes for Near-Infrared Dyes in Capillary Electrophoresis Protein Applications.- Capillary Electrophoresis in the Analysis and Monitoring of Biotechnological Processes.- Capillary Electrophoresis of Proteins in a Quality Control Environment.- Analysis of Neutral N-Linked Oligosaccharides From Antibodies Using Free-Solution Capillary Electrophoresis in Bare Fused-Silica Capillaries.- Affinity Capillary Electrophoresis to Examine Receptor-Ligand Interactions.- Screening Major Binding Sites on Human Serum Albumin by Affinity Capillary Electrophoresis.- Using Charge Ladders and Capillary Electrophoresis to Measure the Charge, Size, and Electrostatic Interactions of Proteins.- Frontal Analysis Continuous Capillary Electrophoresis for Protein-Polyelectrolyte Binding Studies.- Analysis of Proteins by CE, CIEF, and Microfluidic Devices With Whole-Column-Imaging Detection.- Capillary Electrophoresis-Electrospray Ionization Mass Spectrometry of Amino Acids, Peptides, and Proteins.- Capillary Isoelectric Focusing-Mass Spectrometry of Proteins and Protein Complexes.- Integrated System for Rapid Proteomics Analyses Using Microfluidic Devices Coupled to Nanoelectrospray Mass Spectrometry.