HPLC of Peptides and Proteins: Methods and Protocols

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Marie-Isabel Aguilar
Springer Science & Business Media, 2004 - Medical - 413 pages
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Hands-on 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. Follow step-by-step instructions that ensure experimental success Develop your own separation and analytical protocols for peptide and protein analysis.
 

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Contents

HPLC of Peptides and Proteins Basic Theory and Methodology MarieIsabel Aguilar
3
ReversedPhase HighPerformance Liquid Chromatography
9
IonExchange Chromatography
23
HighPerformance Hydrophobic Interaction Chromatography
45
Gel Filtration Chromatography
55
Hydrophilic Interaction Chromatography
75
Immobilized Metal Ion Affinity Chromatography of Proteins
89
Immunoaffinity Chromatography of Proteins
103
Proteolytic Peptide Mapping
227
Mass Spectrometric Characterization of Posttranslationally Modified Proteins Phosphorylation
245
Analyses of Glycopeptides and Glycoproteins by Liquid ChromatographyMass Spectrometry and Liquid ChromatographyTandem Mass Spectrometry
263
HPLC in the Analysis of Peptide Metabolism
275
Isolation and Characterization of Naturally Processed MHCBound Peptides From the Surface of AntigenPresenting Cells
291
Australian FunnelWeb Spider Venom Analyzed With OnLine RPHPLC Techniques
307
HPLC and Mass Spectrometry of Intrinsic Membrane Proteins
323
IgG Purification
341

Liquid ChromatographyMass Spectrometry and Tandem Mass Spectrometry of Peptides and Proteins
111
Capillary Separations
143
Micropreparative HPLC of Peptides and Proteins
165
Multidimensional HPLC Purification of Proteins
177
Analytical HighPerformance Liquid Chromatography
183
PrepSemiprep Separations of Peptides
191
LargeScale Protein Chromatography
211
DNABinding Proteins LCMS to Identify Key Domains in RNA PolymerasePromoter Interactions
351
Sensitive Enzymatic Analysis of Histidine Decarboxylase Using HPLC
365
Automated vs Manual Profiling of Peptide Libraries by Mass Spectrometry
377
Proteome Analysis
387
Index
401
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