This fourth edition volume expands on the previous editions by providing readers with a comprehensive guide to the latest foundational and advanced techniques used by researchers in the field. This book covers the construction of diverse antibody libraries including naïve, synthetic, and immune types and candidate selection using phage, yeast, or mammalian display systems. This book also discusses in silico approaches such as computational design, epitope prediction, and deep learning to improve antibody discovery and characterization. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and authoritative, Antibody Engineering: Methods and Protocols, Fourth Edition is a valuable resource that provides scientists with an extensive toolbox to generate the next generation of antibody-based molecules for research, diagnostics, and therapeutic applications.

Chapter 1 is available Open Access under a Creative Commons Attribution 4.0. International License via link.springer.com

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

IMGT Nomenclature and IMGT Unique Numbering: The Two Pillars of 35 Years of Immunoinformatics for Immunoglobulins (IG) or Antibodies and T Cell Receptors (TR).- Complementarity of Deep Learning and Physics Based Approaches in the Design of New Antibodies.- Prediction of Protein Antigen Epitopes from Monoclonal Antibodies by In Silico Molecular Docking.- Antibody Affinity Maturation by Computational Design.- Construction of Naïve and Immune Human Fab Phage Display Library.- Generation and Panning of a Naïve Human scFv Phage Display Library.- Screening of Monoclonal Antibodies from Integrated Phage and Mammalian Cell Display Libraries.- In Vitro Selection of Antibodies by Ribosome Display.- Yeast Display and Competitive FACS: Powerful Tools for the Selection of Rare Clones from an scFv Library.- Transient Expression and Characterization of Antibodies and Antibody Fragments.- Updates to Guidance of Using Biacore for Protein-Protein Interactions Analysis.- Epitope Binning: Clustering Monoclonal Antibodies on Monomeric and Multivalent Antigens.- High-Throughput Profiling of Antibody-Binding Epitopes Using AbMap.- Ultra-Dilute Developability Analysis of Antibody Self-Association and Non-Specific Binding.- Characterizing pH-Dependent Interactions Between Immunoglobulin and the Neonatal Fc Receptor Using Surface Plasmon Resonance.- Simultaneous Affinity Maturation and Liability Removal from Antibodies.- Antibody Humanization.- Production of Antibody Fragments Suitable for 1:1, Site-Specific Labelling.- Combination of Fc-Protein Engineering Strategies: Design and Evaluation of Antibody Effector Functions.- Systematic Production of IgG-Like Bispecific Antibodies via Intein-Mediated Protein Trans-Splicing.- Generation and Characterization of CAR-T Cells.- Generation and Characterization of Bispecific VHH-Derived Surrogate Agonists Mimicking the Function of IL-18.- Antibody-Drug Conjugates Synthesis Using Maleimide Chemistry.