Epitope Mapping Protocols

This detailed new edition explores methodologies for the characterization of antibody-antigen interactions tailored to different applications and requirements. Beginning with an overview of key concepts and definitions related to antibody epitopes, the book continues with sections on structural biology methods, display technologies to analyze libraries of proteins or protein arrays, techniques based on protein labeling, as well as computational approaches and binding assays that rely on binding interactions or competition. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, Epitope Mapping Protocols, Fourth Edition serves as an ideal resource for scientists from a wide range of fields involved in critical antibody research.

Chapter 19 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

An Overview of Epitopes and Methods for Antibody Epitope Mapping.- Antibody-Antigen Epitope Mapping by X-Ray Crystallography.- Mapping Antibody Epitopes by Solution NMR Spectroscopy: Practical Considerations.- Cryo-Electron Microscopy-Based Epitope Mapping.- Epitope Mapping Using Yeast Display and Next Generation Sequencing.- Rapid Mapping of Protein Binding Sites and Conformational Epitopes by Coupling Yeast Surface Display to Chemical Labeling and Deep Sequencing.- High-Density Antigen Microarrays for the Assessment of Antibody Selectivity.- Mammalian Cell Display with Automated Oligo Design and Library Assembly Allows for Rapid Residue Level Conformational Epitope Mapping.- Epitope Mapping of Antibodies Using Bacterial Cell Surface Display of Gene Fragment Libraries.- Mapping of Antibody Structural Epitopes Using Bacterial Cell Surface Display of Folded Domains.- Highly Multiplexed Immunoassays for the Validation of Anti-GPCR Antibodies.- Probing Antibody Binding Sites on G Protein-Coupled Receptors Using Genetically-Encoded Photo-Activatable Cross-Linkers.- Genetic Code Expansion to Enable Site-Specific Post-Translational Peptide Epitope Tagging of G Protein-Coupled Receptors in Live Cells.- Multiplexed Epitope Mapping of HIV-1 Neutralizing Antibodies by Coupling Chemical Labeling of Cysteines to Deep Sequencing.- Predicting Epitopes of Synthetic Binding Proteins Using Protein Structure Prediction and Protein-Protein Docking.- Structure-Based Epitope Profiling with the Structural Profiling of Antibodies to Cluster by Epitope 2 (SPACE2) Algorithm.- Binding Kinetics and Epitope Binning Using Surface Plasmon Resonance.- High-Throughput Epitope Binning Analysis of Human Monoclonal Antibodies to Assess Epitope Diversity Using the Octet Bio-Layer Interferometry Platform.- Competitive Epitope Binning Using HT-SPR.- Label-Free Cell-Based Assay for Characterization of Biomolecules and Receptors.- B Cell Epitope Mapping by High Density Competition ELISAs.