Synthetic Peptides As Antigens

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ISBN: 9780444821768 | 0444821767
Cover: Paperback
Copyright: 11/19/1999

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This newest edition to the Laboratory Techniques Series gives current state of the art use of synthetic peptides in molecular biology and practical protocols on how to conjugate peptides, immunize animals with peptides and monitor immune responses to peptides in vitro. It gives background information on antigenic specificity, prediction of antigenic sites in proteins and applications of peptides in immunology and virology, as probes in diagnosis and as vaccines. The book also describes antigenicity of proteins and methods to localize antigenic sites as well as methods for predicting epitoxes, and gives detailed protocols for peptide-carrier conjugation, immunization with peptides, and peptide immunoassays. The volume also describes typical use of antipeptide antibodies in molecular and cellular biology as well as the use of peptides in the diagnosis of viral infections and autoimmune diseases, and the use of peptides as potential synthetic vaccines. An excellent edition to an excellent series, available in hardbound and paperback.

Acknowledgements

List of abbreviations

vii

Molecular dissection of protein antigens and the prediction of epitopes

(78)

M.H.V. Van Regenmortel

Introduction

(2)

Definition of antigenicity and the concept of epitope

(7)

Types of epitopes

(7)

Methods used for localizing epitopes

(22)

Crystallographic analysis of antigen-antibody complexes

(7)

Binding studies with analogues and mutagenized molecules

(3)

Binding studies with peptide fragments

(3)

Synthetic peptides as antigenic probes

(3)

Peptide sequences inserted in recombinant proteins

(1)

Binding studies with antipeptide antibodies

(2)

Differential sensitivity of free and bound epitopes to chemical attack or amide exchange

(1)

Competitive topographic mapping

(1)

The antigenic structure of model proteins

(14)

Myoglobin

(6)

Lysozyme

(2)

Tobacco mosaic virus protein

(6)

Antigenicity prediction

(15)

General principles

(5)

Review of propensity scales

(1)

Hydrophilicity scale of Hopp and Woods (1981)

(1)

Hydropathy scale of Kyte and Doolittle (1982)

(1)

Hydrophilicity scale of Parker et al. (1986)

(2)

Acrophilicity scale of Hopp (1984)

(1)

Flexibility scale of Karplus and Schulz (1985)

(1)

Antigenicity scale of Welling et al. (1985)

(1)

Antigenicity index of Jameson and Wolf (1988)

(1)

Turn scales of Pellequer et al. (1993)

(1)

Comparative efficacies of different scales

(5)

T-cell epitopes

(11)

Nature of T-cell epitopes

(3)

The peptide-binding groove of MHC class I and II molecules

(3)

T-cell receptor binding of peptide-MHC complexes

(2)

Prediction of T-cell epitopes

(3)

Peptide-carrier conjugation

(54)

S. Muller

Introduction

(1)

Choice of carrier

(3)

Optimal peptide density on carrier protein

(2)

Point of attachment on peptide chain

(1)

Chemical coupling

(24)

Glutaraldehyde

(1)

Reaction mechanism

(2)

Procedure (one-step method)

(1)

Bisimido esters

(1)

Reaction mechanism

(1)

Procedure used in the case of dimethyl suberimidate (one-step method)

(2)

Carbodiimides

(1)

Reaction mechanism

(1)

Procedure (Goodfriend et al., 1964)

(1)

Bis-diazobenzidine

(1)

Reaction mechanism

(1)

Procedure (Briand et al., 1985; Tamura and Bauer, 1982)

(1)

m-Maleimido benzoyl-N-hydroxysuccinimide ester (MBS)

(1)

Reaction mechanism

(2)

Procedure (two-step method)

(1)

N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP)

(1)

Reaction mechanism

(2)

Procedure (two-step method)

100

(2)

Imidoesters: 2-iminothiolane or 2-iminotetrahydrothiophene

102

(1)

Reaction mechanism

102

(1)

Procedure (one-step method)

103

(1)

Hydrazone formation following periodate oxidation of N-terminal serine on threonine residues

104

(1)

Reaction mechanism

104

(1)

Procedure (two-step method)

105

(3)

Other chemical coupling agents

108

(1)

Photochemical coupling

109

(5)

Reaction mechanism

109

(4)

Procedure

113

(1)

Coupling of peptides of liposomes

114

(5)

Reaction mechanism

114

(2)

Procedure (Frisch et al., 1991)

116

(1)

Synthesis of N-[4-(p-maleimidophenyl)butyryl] phosphatidylethanolamine (MPB-PE)

116

(1)

Preparation of large and small unilamellar vesicles and peptide coupling

117

(1)

Preparation of encapsulated peptide

118

(1)

Lyophilized liposomes (Friede et al., 1993c)

119

(1)

Coupling of peptides to solid supports

119

(7)

General remarks

119

(1)

Procedure using iodoacetyl activated for immobilization of peptides through sulfhydryl groups

120

(1)

Reaction mechanism

120

(1)

Procedure

121

(5)

Determination of peptide: carrier ratio of conjugates

126

(3)

Peptide derivatization

129

(4)

Reversible protection of amino groups with citraconic anhydride

129

(1)

Biotinylation of peptides

130

(3)

Immunization with peptides

133

(46)

S. Muller

Introduction

133

(1)

The Choice of animal

134

(2)

The immunogen

136

(22)

Free versus conjugated peptides

136

(4)

Immunization with conjugated peptides

140

(4)

Possible experimental pitfalls due to the generation of antibodies to the coupling agent used for peptide conjugation

144

(1)

Use of liposomes as peptide vehicles

145

(4)

Synthetic branched peptides

149

(3)

Peptide construction involving B-and T-epitopes

152

(6)

The adjuvant

158

(9)

The route of injection

167

(2)

Specific immunization protocols

169

(8)

Method of Walter et al. (1980); Patschinsky et al. (1984)

170

(1)

Method of Green et al. (1982)

170

(1)

Method of Tanaka et al. (1985)

171

(1)

Method of Muller et al. (1986) and Dumortier et al. (1998)

171

(1)

Method of Nussberger et al. (1985)

171

(1)

Method of Choppin et al. (1986)

172

(2)

Method of Young et al. (1993)

174

(1)

Method of De Boer et al. (1987)

175

(1)

Method of Lu et al. (1991) with MAP

175

(1)

Method of Tuchscherer et al. (1992) with TASP

176

(1)

Concluding remarks

177

(2)

Peptide immunoassays

179

(36)

M. H. V Van Regenmortel

Introduction

179

(2)

Types of solid-phase immunoassays

181

(15)

The pepscan technique

181

(5)

ELISA

186

(10)

Solid-phase immunoassay procedures

196

(6)

Indirect ELISA using immobilized peptide

196

(2)

Double antibody sandwich assay

198

(4)

Solid-phase radioimmunoassay (RIA)

202

(1)

Dot immunobinding assay

202

(1)

Spotscan assay

203

(1)

Biosensor assays

204

(6)

Description of the BIAcore instrument

204

(2)

Analysis of peptide-antibody interactions with BIAcore

206

(3)

Immobilization of peptides on sensor chips

209

(1)

Measurement of affinity constants

210

(2)

Monitoring of the immune response to peptides

212

(3)

Use of antipeptide antibodies in molecular and cellular biology

215

(22)

S. Muller

Detection of gene products with antipeptide antibodies

215

(17)

Detection of putative proteins on the basis of nucleic acid sequences

217

(3)

Antipeptide antibody probes for structural and functional studies of gene products

220

(1)

Immunodetection of in vitro translation products using antipeptide antibodies

220

(1)

Selected techniques used for the immunological detection of gene products

221

(1)

Gene isolation with antibody probes using &lembda;gt11 expression vector

222

(3)

Immunochemical detection of protein related to the human c-myc exon 1

225

(2)

Use of antipeptide antibodies for the immunodetection of plant viral nonstructural proteins

227

(4)

General comments

231

(1)

Use of antipeptide antibodies in immunohistochemistry and immunocytochemistry

232

(5)

General considerations

232

(1)

Examples of procedures

233

(4)

The use of peptides for diagnosing viral infections

237

(10)

M. H. V. Van Regenmortel

Mimicry of viral epitopes with synthetic peptides

237

(5)

Synthetic peptides for viral diagnosis

242

(2)

Peptide-based immunoassays

244

(3)

Peptides in diagnosis of autoimmune diseases

247

(34)

S. Muller

Introduction

247

(1)

Methods of detection and quantification of autoantibodies with synthetic peptides

248

(14)

Epitope mapping with synthetic peptides

248

(5)

Respective merits and limitations of the different approaches used to delineate epitopes recognized by autoantibodies

253

(5)

Cross-reactivity of autoantibodies with synthetic peptides and the cognate protein

258

(4)

Specific examples of autoepitope mapping data

262

(11)

B-cell epitopes of Ro60 protein

263

(2)

B-cell epitopes of SmD1 protein

265

(5)

B-cell epitopes of U1C protein

270

(3)

Prediction of epitopes recognized by autoantibodies

273

(5)

Peptides mimicking sites of post-translational modification recognized by autoantibodies

278

(2)

Concluding remarks

280

(1)

Synthetic peptides as vaccines

281

(38)

M. H. V. Van Regenmortel

Introduction

281

(3)

Antiviral vaccines

284

(26)

Foot-and-mouth disease virus

284

(10)

Poliovirus

294

(4)

Influenza virus

298

(4)

Canine parvovirus

302

(3)

Measles virus

305

(2)

Human immunodeficiency virus

307

(3)

Other viruses

310

(1)

Vaccines against bacterial infections

310

(1)

Vaccines against parasites

311

(2)

Are molecular design strategies applicable to the development of synthetic vaccines?

313

(1)

Empirical discovery rather than molecular design will bring about new synthetic vaccines

314

(5)

References

319

(56)

Subject index

375

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Synthetic Peptides As Antigens

Summary

Table of Contents

Supplemental Materials