Aguilar M.I. (ed.) Methods in molecular biology. Volume 251. HPLC of Peptides and Proteins. Methods and protocols

Humana Press Inc., 2004. — 411 p.This volume opens with an overview of the basic theory and general methodology of HPLC, with particular reference to the key separation parameters that can be manipulated to achieve high resolution. Each of the commonly used HPLC techniques are covered in Chapters 2–9, whereas methods for capillary to large-scale preparative isolation are described in Chapters 10–
15. Chapters 16–27 provide those already experienced in HPLC with a number of specific applications, as in 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 modifications, neuropeptide processing, glycopeptides and glycoproteins, MHC-binding peptides, toxins/venoms, membrane proteins, antibodies, combinatorial and proteome analysis, and enzymatic activity. HPLC of Peptides and Proteins: Methods and Protocols will be a valuable resource for a wide range of scientists, including biochemists, molecular biologists, pharmacologists, and microbiologists, who work with peptides and/or proteins in both academic and biotechnology laboratories.Modes ofHPLC
HPLC of Peptides and Proteins
Basic Theory and Methodology
Reversed-Phase High-Performance Liquid Chromatography on-Exchange Chromatography
High-Performance Hydrophobic Interaction Chromatography
Gel Filtration Chromatography
Hydrophilic Interaction Chromatography
Immobilized Metal Ion Affinity Chromatography of Proteins
mmunoaffinity 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
lgG Purification
DNA–Binding Proteins: LC–MS to Identify Key Domains
In RNA Polymerase–Promoter Interactions
Sensitive Enzymatic Analysis of Histidine Decarboxylase
Using HPLC
Automated vs Manual Profiling of Peptide Libraries by Mass Spectrometry
Proteome Analysis