Interest in epitope mapping, or finding out where antibodies bind to their antigens, is by no means restricted to immunologists, but is shared by biolo gists from a wide range of disciplines in which antibodies are used as molecu lar reagents. The epitope mapper may be interested in studying protein-protein interactions, in developing an immunoassay, in producing protective peptide vaccines, in investigating the pathogenesis of autoimmune diseases, or in defining protein topology in intact cells or organelles, to mention only a few of the possibilities. The aim of Epitope Mapping Protocols is to provide both a useful range of alternative practical methods for the experienced mapper and a fairly com prehensive introduction for someone embarking on antibody production and mapping for the first time. Contributors were encouraged to illustrate their protocols with results from their own research and most of them elected to do so. After an introductory chapter, the protocols are arranged in three groups: The first group of twelve methods uses (or can use) whole, native antigens and may therefore be suitable for conformational epitopes; the second group of five uses peptides or peptide libraries; and the third group of eleven requires antigen expressed from recombinant DNA. Within this last group, methods more likely applicable to conformational epitopes are placed first.

Inhalt
Overview.- Crystallographic Studies of Antigen-Antibody Interactions.- Epitope Mapping Antibody-Antigen Complexes by Nuclear Magnetic Resonance Spectroscopy.- Mapping Epitopes on Antigens by Immunodiffusion in Gel.- A Simple Solid-Phase Competition Assay with Labeled Antigen.- Epitope Mapping by Antibody Competition.- Epitope Mapping by Surface Plasmon Resonance in the BIAcore.- Identifying Residues in Antigenic Determinants by Chemical Modification.- Epitope Mapping by Differential Chemical Modification of Antigens.- Epitope Mapping by Proteolysis of Antigen-Antibody Complexes.- Proteolytic Fragmentation for Epitope Mapping.- Epitope Mapping by Chemical Fragmentation.- Probing Antibody-Antigen Interactions by Mass Spectrometry.- Epitope Mapping Using Multipin Peptide Synthesis.- SPOT Synthesis.- Tea Bag Synthesis of Positional Scanning Synthetic Combinatorial Libraries and Their Use for Mapping Antigenic Determinants.- Epitope Mapping Using Phage-Displayed Peptide Libraries.- Reiterative Screening of Phage-Display Peptide Libraries with Antibodies.- Homolog Scanning.- Epitope Mapping Using an Oligonucleotide Replacement Method.- Epitope Mapping by Region-Specified PCR Mutagenesis.- Random Fragment Libraries Using Yeast Expression Plasmid.- Epitope Mapping Using Random Fragment Expression Libraries in ? Phages.- Random Fragment Libraries Displayed on Filamentous Phage.- Epitope Mapping by Expression of Restriction Enzyme or PCR Fragments in Bacterial Plasmids.- Epitope Mapping of Protein Antigens by Expression-PCR (E-PCR).- Epitope Mapping on Extracellular Domains of Cell-Surface Proteins Using Exonuclease III.- An Improved Method for Mapping Epitopes of Recombinant Antigens by Transposon Mutagenesis.- Incomplete Polypeptides of In Vitro Translation for EpitopeLocalization.- T-Cell Epitope Mapping with Synthetic Peptides and Peripheral Blood Mononuclear Cells.- Use of Natural or Selected Mutants and Variants for Epitope Mapping.- Production of Panels of Monoclonal Antibodies by the Hybridoma Method.- Production of Phage-Display Antibodies for Epitope Mapping.