Bioconjugation: Methods and Protocols

This book explores well-established and emerging conjugation strategies that are relevant for proteins used in the field of precision medicine, focusing on techniques that are suitable for antibodies, antibody-fragments such as Fabs, scFvs, or nanobodies, scaffold proteins such as FN3 or DARPin, peptides, or model proteins. Although centered on the development of bioconjugates rather than their application, most protocols also show the conjugation of the targeting vehicle to a diagnostic or therapeutic entity, with the end-product most often being an antibody-drug conjugate, an optical probe, a nanomedicine, or a radiopharmaceutical. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.  Authoritative and practical, Bioconjugation: Methods and Protocols is an ideal guide for researchers looking toward precision medicine in order to expand the vital field of drug discovery.

Table of Contents:
Site-Specific Conjugation of Thiol-Reactive Cytotoxic Agents to Non-Native Cysteines of Engineered Monoclonal Antibodies.- Application of Next Generation Maleimides (NGMs) to Site-Selective Antibody Conjugation.- Lysine Bioconjugation on Native Albumin with a Sulfonyl Acrylate Reagent.- Engineering Dual-Variable-Domains for the Generation of Site-Specific Antibody-Drug Conjugates.- Efficient Production of Homogeneous Lysine-Based Antibody Conjugates Using Microbial Transglutaminase.- Engineering Antibodies with C-Terminal Sortase-Mediated Modification for Targeted Nanomedicine.- CRISPR/Cas9-Mediated Genetic Engineering of Hybridomas for Creation of Antibodies that Allow Site-Specific Conjugation.- Site-Specific Modification of Proteins via Trypsiligase.- Site-Selective Functionalization of Nanobodies Using Intein-Mediated Protein Ligation for Innovative Bioconjugation.- Site-Specific Bioconjugation Using SMARTagâ Technology: A Practical and Effective Chemoenzymatic Approach to Generate Antibody-Drug Conjugates.- Protein Labeling and Bioconjugation Using N-Myristoyltransferase.- One-Step Fluorescent Protein Labeling by Tubulin Tyrosine Ligase.- Enzymatically Catalyzed Radiofluorination of Biomolecules.- Site-Selective Enzymatic Labeling of Designed Ankyrin Repeat Proteins Using Protein Farnesyltransferase.- Homogeneous Antibody-Drug Conjugates via Glyco-Engineering.- Labeling Proteins at Site-Specifically Incorporated 5-Hydroxytryptophan Residues Using a Chemoselective Rapid Azo-Coupling Reaction.- Facile Site-Specific Multi-Conjugation Strategies in Recombinant Proteins Produced in Bacteria.- Site-Specific Photocrosslinking to Immunoglobulin G Using Photoreactive Antibody-Binding Domains.- SpyTag-SpyCatcher Chemistry for Protein Bioconjugation In Vitro and Protein Topology Engineering In Vivo.- FN3 Protein Conjugates for Cancer Diagnosis and Imaging Studies.