Epitope Mapping Protocols

This third edition volume expands on the previous editions with more detailed research on the characterization of antibody antigen interactions between different users with different requirements. The chapters in this book are divided into four parts: Part One looks at the entire native antigen and covers traditional structural biology techniques such as nuclear magnetic resonance and x-ray crystallography. Part Two talks about protein fragments derived from antigens, and discusses binding regions within antigen sequence using bacterial surface display and ELISA, for example. Part Three describes the use of surface plasmon resonance spectroscopy and biolayer interferometry, and Part Four highlights methods used to identify new antigens and assess antibody cross-reactivity. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readilyreproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Thorough and cutting-edge, Epitope Mapping Protocols, Third Edition is a valuable resource for anyone interested in furthering their research in this expanding field.

An Introduction to Epitope Mapping.- Epitope Mapping of Antibody-Antigen Interactions with X-Ray Crystallography.- Mapping Antibody Epitopes by Solution NMR Spectroscopy: Practical Considerations.- Label-Free Cell-Based Assay for Characterization of Biomolecules and Receptors.- Probing Antibody Binding Sites on G Protein-Coupled Receptors Using Genetically-Encoded Photo-Activatable Cross-Linkers.- Rapid Mapping of Protein Binding Sites and Conformational Epitopes by Coupling Yeast Surface Display to Chemical Labeling and Deep Sequencing.- Epitope Mapping Using Yeast Display and Next Generation Sequencing.- B-Cell Epitope Mapping Using a Library of Overlapping Synthetic Peptides in an Enzyme-Linked Immunosorbent Assay.- Array-In-Well Epitope Mapping of Phage-Displayed Antibodies.- Epitope Mapping of Antibodies Using Bacterial Cell Surface Display of Gene Fragment Libraries.- Discontinuous Epitope Mapping of Antibodies Using Bacterial Cell Surface Display of Folded Domains.- Kinetic Analysis and Epitope Binning Using Surface Plasmon Resonance.- Epitope Binning of Human Monoclonal Antibodies in Classical Sandwich and In-Tandem Orientation Using the Octet System Based on Biolayer Interferometry.- Antibody Specificity Profiling Using Protein Microarrays.- High-Density Antigen Microarrays for Assessment of Antibody Selectivity and Off-Target Binding.- Multiplexed Antigen Bead Arrays for Assessment of Antibody Selectivity and Epitope Mapping.- Antibody Biomarker Discovery Via In Vitro Evolution of Peptides.- Bioinformatics Tools for the Prediction of T Cell Epitopes.