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HPLC of Peptides and Proteins

Methods and Protocols

Series: Methods in Molecular Biology

by Marie-Isabel Aguilar

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Main description:

The introduction of high-performance liquid chromatography (HPLC) to the analysis of peptides and proteins some 25 years ago revolutionized the biological sciences by enabling the rapid and sensitive analysis of peptide and protein structure through the exquisite speed, sensitivity, and resolution that can be easily obtained. Today, HPLC in its various modes has become the pivotal technique in the characterization of peptides and proteins and currently plays a critical role in both our understanding of biological processes and in the development of peptide- and protein-based pharmaceuticals. The number of applications of HPLC in peptide and protein purification continues to expand at an extremely rapid rate. Solid-phase peptide synthesis and recombinant DNA techniques have allowed the production of large quantities of peptides and proteins that need to be highly purified. HPLC techniques are also used extensively in the isolation and characterization of novel proteins that will become increasingly important in the postgenomic age. The design of multidimensional purification schemes to achieve high levels of product purity further demonstrates the power of HPLC techniques not only in the characterization of cellular events, but also in the production of pepti- and protein-based therapeutics. HPLC continues to be at the heart of the analytical techniques with which scientists in both academia and in industry must arm themselves to be able to fully characterize the identity, purity, and potency of peptides and proteins.

Contents:

Part I. Modes of HPLC

HPLC of Peptides and Proteins: Basic Theory and Methodology
Marie-Isabel Aguilar

Reversed-Phase High-Performance Liquid Chromatography
Marie-Isabel Aguilar

Ion-Exchange Chromatography
Peter Stanton

High-Performance Hydrophobic Interaction Chromatography
Kalman Benedek

Gel Filtration Chromatography
Peter Stanton

Hydrophilic Interaction Chromatography
Herbert Lindner and Wilfried Helliger

Immobilized Metal Ion Affinity Chromatography of Proteins
Michael Zachariou

Immunoaffinity Chromatography of Proteins
Stuart R. Gallant

Liquid Chromatography-Mass Spectrometry and Tandem Mass Spectrometry of Peptides and Proteins
Tun-Li Shen and Kathleen R. Noon

Part II. Preparative Methodologies

Capillary Separations
Montserrat Carrascal and Joaquin Abian

Micropreparative HPLC of Peptides and Proteins
Edouard C. Nice and Marie-Isabel Aguilar

Multidimensional HPLC Purification of Proteins
Edouard C. Nice and Marie-Isabel Aguilar

Analytical High-Performance Liquid Chromatography
Kalman Benedek

Prep/Semiprep Separations of Peptides
Denis B. Scanlon and James Finlayson

Large-Scale Protein Chromatography
Joseph Bertolini, Peter Gomme, and Patrick Thomas

Part III. Applications

Proteolytic Peptide Mapping, Peter Hojrup

Mass Spectrometric Characterization of Posttranslationally Modified Proteins-Phosphorylation
Martin R. Larsen

Analyses of Glycopeptides and Glycoproteins by Liquid Chromatography-Mass Spectrometry and Liquid Chromatography-Tandem Mass Spectrometry
Nana Kawasaki, Miyako Ohta, Satsuki Itoh, and Takao Hayakawa

HPLC in the Analysis of Peptide Metabolism
Rebecca A. Lew

Isolation and Characterization of Naturally ProcessedMHC-Bound Peptides From the Surface of Antigen-Presenting Cells
Anthony W. Purcell

Australian Funnel-Web Spider Venom Analyzed With On-Line RP-HPLC Techniques
David Wilson and Paul Alewood

HPLC and Mass Spectrometry of Intrinsic Membrane Proteins
Julian P. Whitelegge

IgG Purification
Maree S. Powell and Bruce D. Wines

DNA-Binding Proteins: LC-MS to Identify Key Domains in RNA Polymerase-Promoter Interactions
Malcolm Buckle

Sensitive Enzymatic Analysis of Histidine Decarboxylase Using HPLC
Jordi Gomez-Ramirez, Isaac Blanco, and Jordi Ortiz

Automated vs Manual Profiling of Peptide Libraries by Mass Spectrometry
Jean-Louis Aubagnac, Christine Enjalbal, Jean Martinez, Pierre Sanchez, and Gilles Subra

Proteome Analysis
Matthew J. Powell and Aaron T. Timperman

PRODUCT DETAILS

ISBN-13: 9781617372865
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 428
Weight: 626g
Availability: Available
Subcategories: Biochemistry

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