Synthetic Peptides As Antigens

by Muller; van Regenmortel

• ISBN13: 9780444821768
• ISBN10: 0444821767
• Edition: 1st
• Format: Paperback
• Copyright: 1999-11-19
• Publisher: Elsevier Science

Summary

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.

Table of Contents

Acknowledgements	v
List of abbreviations	vii
Molecular dissection of protein antigens and the prediction of epitopes	1	(78)
M.H.V. Van Regenmortel
Introduction	1	(2)
Definition of antigenicity and the concept of epitope	3	(7)
Types of epitopes	10	(7)
Methods used for localizing epitopes	17	(22)
Crystallographic analysis of antigen-antibody complexes	19	(7)
Binding studies with analogues and mutagenized molecules	26	(3)
Binding studies with peptide fragments	29	(3)
Synthetic peptides as antigenic probes	32	(3)
Peptide sequences inserted in recombinant proteins	35	(1)
Binding studies with antipeptide antibodies	36	(2)
Differential sensitivity of free and bound epitopes to chemical attack or amide exchange	38	(1)
Competitive topographic mapping	39	(1)
The antigenic structure of model proteins	39	(14)
Myoglobin	39	(6)
Lysozyme	45	(2)
Tobacco mosaic virus protein	47	(6)
Antigenicity prediction	53	(15)
General principles	53	(5)
Review of propensity scales	58	(1)
Hydrophilicity scale of Hopp and Woods (1981)	58	(1)
Hydropathy scale of Kyte and Doolittle (1982)	59	(1)
Hydrophilicity scale of Parker et al. (1986)	59	(2)
Acrophilicity scale of Hopp (1984)	61	(1)
Flexibility scale of Karplus and Schulz (1985)	61	(1)
Antigenicity scale of Welling et al. (1985)	62	(1)
Antigenicity index of Jameson and Wolf (1988)	62	(1)
Turn scales of Pellequer et al. (1993)	63	(1)
Comparative efficacies of different scales	63	(5)
T-cell epitopes	68	(11)
Nature of T-cell epitopes	68	(3)
The peptide-binding groove of MHC class I and II molecules	71	(3)
T-cell receptor binding of peptide-MHC complexes	74	(2)
Prediction of T-cell epitopes	76	(3)
Peptide-carrier conjugation	79	(54)
S. Muller
Introduction	79	(1)
Choice of carrier	80	(3)
Optimal peptide density on carrier protein	83	(2)
Point of attachment on peptide chain	85	(1)
Chemical coupling	85	(24)
Glutaraldehyde	85	(1)
Reaction mechanism	85	(2)
Procedure (one-step method)	87	(1)
Bisimido esters	88	(1)
Reaction mechanism	88	(1)
Procedure used in the case of dimethyl suberimidate (one-step method)	89	(2)
Carbodiimides	91	(1)
Reaction mechanism	91	(1)
Procedure (Goodfriend et al., 1964)	92	(1)
Bis-diazobenzidine	93	(1)
Reaction mechanism	93	(1)
Procedure (Briand et al., 1985; Tamura and Bauer, 1982)	94	(1)
m-Maleimido benzoyl-N-hydroxysuccinimide ester (MBS)	95	(1)
Reaction mechanism	95	(2)
Procedure (two-step method)	97	(1)
N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP)	98	(1)
Reaction mechanism	98	(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