Modes of HPLC -- HPLC of Peptides and Proteins -- Reversed-Phase High-Performance Liquid Chromatography -- Ion-Exchange Chromatography -- High-Performance Hydrophobic Interaction Chromatography -- Gel Filtration Chromatography -- Hydrophilic Interaction Chromatography -- Immobilized Metal Ion Affinity Chromatography of Proteins -- Immunoaffinity Chromatography of Proteins -- Liquid Chromatography-Mass Spectrometry and Tandem Mass Spectrometry of Peptides and Proteins -- Preparative Methodologies -- Capillary Separations -- Micropreparative HPLC of Peptides and Proteins -- Multidimensional HPLC Purification of Proteins -- Analytical High-Performance Liquid Chromatography -- Prep/Semiprep Separations of Peptides -- Large-Scale Protein Chromatography -- Applications -- Proteolytic Peptide Mapping -- Mass Spectrometric Characterization of Posttranslationally Modified Proteins-Phosphorylation -- Analyses of Glycopeptides and Glycoproteins by Liquid Chromatography-Mass Spectrometry and Liquid Chromatography-Tandem Mass Spectrometry -- HPLC in the Analysis of Peptide Metabolism -- Isolation and Characterization of Naturally Processed MHC-Bound Peptides From the Surface of Antigen-Presenting Cells -- Australian Funnel-Web Spider Venom Analyzed With On-Line RP-HPLC Techniques -- HPLC and Mass Spectrometry of Intrinsic Membrane Proteins -- IgG Purification -- DNA-Binding Proteins -- Sensitive Enzymatic Analysis of Histidine Decarboxylase Using HPLC -- Automated vs Manual Profiling of Peptide Libraries by Mass Spectrometry -- Proteome Analysis.

High performance liquid chromatography (HPLC) plays a critical role today in both our understanding of biological processes and in the development of peptide and protein-based pharmaceuticals. In HPLC of Peptides and Proteins: Methods and Protocols, leading experts from academia and industry comprehensively describe how to successfully perform all the critical HPLC techniques needed for the analysis of peptides and proteins. The methods range from commonly used techniques to those for capillary to large-scale preparative isolation. The authors have also presented a number of specific applications as case studies to illustrate the analytical approaches to a particular separation or assay challenge, with examples drawn from contemporary fields in biochemistry and biotechnology. These applications include proteolytic mapping, posttranslational modification, neuropeptide processing, glycopeptides and glycoproteins, MHC-binding peptides, toxins/venoms, membrane proteins, antibodies, combinatorial and proteome analysis, and enzymatic activity. Each readily reproducible protocol contains background notes, step-by-step instructions, reagent and equipment lists, and tips on both troubleshooting and avoiding known pitfalls. Comprehensive and easy-to-follow, HPLC of Peptides and Proteins: Methods and Protocols offers every biologist and biotechnologist at all levels of expertise ready access to the powerful tools needed to develop and carry out peptide and protein separations and analyses today.