High-Performance Liquid Chromatography of Peptides and Proteins

Separation, Analysis, and Conformation

Colin T. Mant, Robert S. Hodges, Colin T. Mant, Robert S. Hodges

• Publisher: CRC Press
• Year: 2017
• Print ISBN:
  9780849365492
• eBook ISBN:
  9781351440745
• Topic: Medicine
• Subtopic: Analytic Chemistry

This book consists of a series of 82 precise, easy-to-read articles by internationally renowned scientists and emphasizes the practical approach to HPLC with minimal theory, although the underlying principles for peptide and protein separations are clearly expressed. All of the major modes of microbore, ultrafast and analytical HPLC are discussed, including size-exclusion, ion-exchange, reversed-phase, hydrophobic interaction, and affinity and immunoaffinity chromatography. A section on preparative HPLC, including displacement techniques, is also presented. Problem-solving approaches to the separation of various classes of biologically active peptides and proteins are thoroughly explored, while the importance of peptide standards for monitoring column performance and for optimizing separation conditions is emphasized. Several articles focus on the choice of the correct detection method (electrochemical, UV, fluorescence), as well as the need for a proper knowledge of approaches to column and instrument maintenance and trouble-shooting. A section on predictive approaches deals with both computer simulation of peptide separations and peptide structure. The book also includes complementary techniques to HPLC, as well as other useful applications of HPLC. It enables both novice and experienced chromatographers to realize the full potential of this extremely powerful technique, in the process making an important contribution to scientific literature.

Indexes

STRUCTURE INDEX
Amino Acids Found in Proteins (Including Three-Letter and One-Letter Codes)

Amino Acids not Found in Proteins

General Structures

INDEX

A

Absorbance ratios, 615, 617–618

Absorption maximum, 309

Accessible silanol sites, 411

Acetonitrile, 129–131, 292, 298, 307, 310, 327, 411, 415, 439–440, 442, 644–645, 680–682, 692, 710–711, 773–775, 780, 784, 793, 825, 837

cation-exchange chromatography, 176–177

ion-exchange chromatography, use in, 14

Acetylcholine-receptor, 231

Acid elution, 513

Acid proteins, 239

Acid-stable basic proteins, 243

Acidic buffers, 514

Acidic fibroblast growth factor, 501–505

Acidic proteins

pI values, 246

polypeptide standards, 240

separation by cation-exchange high-performance liquid chromatography, 239–244

ACTH, 320–321

Actin, 485

Actin peptide, 485

Active enzyme preparations, 497

Acylation, 890

Additives

ion-exchange chromatography, 211–212

reversed-phase columns, 279

Adenovirus, 226

Adrenal, 199

Adsorbed state, 613

Adsorption behavior, 809–812

Affinity chromatography, 73

advantages of high-performance affinity chromatography over, 481

LC columns, 31

membrane proteins, 234

Affinity procedures

Factor XII, 493–494, 497

plasma kallikrein, 493–494, 497

Aggregation, 233–234, 282, 590

Air bubbles

liquid chromatography detector, 28

liquid chromatography pump, 25

Alanine, 571

Albumin variants, 844

Aldolase, 841

Alkyl chain length, 73

n-Alkyl chain, 274

Alkylphenones, 3, ...

Table of contents

• Cover
• Title Page
• Copyright Page
• Table of Contents
• SECTION I: PRACTICAL ASPECTS OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
• SECTION II: THEORETICAL ASPECTS OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
• SECTION III: SIZE-EXCLUSION CHROMATOGRAPHY
• SECTION IV: ION-EXCHANGE CHROMATOGRAPHY
• SECTION V: REVERSED-PHASE CHROMATOGRAPHY
• SECTION VI: HYDROPHOBIC INTERACTION CHROMATOGRAPHY
• SECTION VII: AFFINITY AND IMMUNOAFFINITY CHROMATOGRAPHY
• SECTION VIII: COLUMN PACKING TECHNIQUES
• SECTION IX: METHODS OF DETECTION
• SECTION X: ANALYSIS OF PEPTIDE AND PROTEIN CONFORMATION BY HPLC
• SECTION XI: INTRODUCTION TO NARROW BORE, MICROBORE, AND RAPID HPLC ANALYSIS TECHNIQUES
• SECTION XII: PREDICTION AND COMPUTER SIMULATION OF PEPTIDE AND PROTEIN SEPARATIONS AND STRUCTURE FOR RESEARCH AND TEACHING
• SECTION XIII: PREPARATIVE CHROMATOGRAPHY OF PEPTIDES AND PROTEINS
• SECTION XIV: ANALYSIS OF PEPTIDES AND PROTEINS
• INDEXES