Main description:

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.

Contents:

1. Site-Specific Conjugation of Thiol-Reactive Cytotoxic Agents to Non-Native Cysteines of Engineered Monoclonal Antibodies

Stephan Dickgiesser, Roland Kellner, Harald Kolmar, and Nicolas Rasche

2. Application of Next Generation Maleimides (NGMs) to Site-Selective Antibody Conjugation

Mauricio Morais, Nafsika Forte, Vijay Chudasama, and James R. Baker

3. Lysine Bioconjugation on Native Albumin with a Sulfonyl Acrylate Reagent

Maria J. Matos, Gonzalo Jimenez-Oses, and Goncalo J.L. Bernardes

4. Engineering Dual-Variable-Domains for the Generation of Site-Specific Antibody-Drug Conjugates

Alex R. Nanna and Christoph Rader

5. Efficient Production of Homogeneous Lysine-Based Antibody Conjugates Using Microbial Transglutaminase

Jared L. Spidel and Earl F. Albone

6. Engineering Antibodies with C-Terminal Sortase-Mediated Modification for Targeted Nanomedicine

Rania A. Hashad, Jaclyn L. Lange, Natasha C.W. Tan, Karen Alt, and Christoph E. Hagemeyer

7. CRISPR/Cas9-Mediated Genetic Engineering of Hybridomas for Creation of Antibodies that Allow Site-Specific Conjugation

Makan Khoshnejad, Jacob S. Brenner, Hamideh Parhiz, and Vladimir R. Muzykantov

8. Site-Specific Modification of Proteins via Trypsiligase

Sandra Liebscher and Frank Bordusa

9. Site-Selective Functionalization of Nanobodies Using Intein-Mediated Protein Ligation for Innovative Bioconjugation

Geert-Jan Graulus, Duy Tien Ta, Huong Tran, Rebekka Hansen, Brecht Billen, Erik Royackers, Jean-Paul Noben, Nick Devoogdt, Serge Muyldermans, Wanda Guedens, and Peter Adriaensens

10. Site-Specific Bioconjugation Using SMARTaga Technology: A Practical and Effective Chemoenzymatic Approach to Generate Antibody-Drug Conjugates

Junjie Liu, Robyn M. Barfield, and David Rabuka

11. Protein Labeling and Bioconjugation Using N-Myristoyltransferase

Karin F.K. Ejendal, Julia G. Fraseur, and Tamara L. Kinzer-Ursem

12. One-Step Fluorescent Protein Labeling by Tubulin Tyrosine Ligase

Dominik Schumacher, Heinrich Leonhardt, Christian P.R. Hackenberger, and Jonas Helma

13. Enzymatically Catalyzed Radiofluorination of Biomolecules

Christopher R. Drake, Michael J. Evans, and Henry F. VanBrocklin

14. Site-Selective Enzymatic Labeling of Designed Ankyrin Repeat Proteins Using Protein Farnesyltransferase

Yi Zhang, Shelby Auger, Jonas V. Schaefer, Andreas Pluckthun, and Mark D. Distefano

15. Homogeneous Antibody-Drug Conjugates via Glyco-Engineering

Feng Tang, Wei Shi, and Wei Huang

16. Labeling Proteins at Site-Specifically Incorporated 5-Hydroxytryptophan Residues Using a Chemoselective Rapid Azo-Coupling Reaction

Partha Sarathi Addy, Sarah B. Erickson, James S. Italia, and Abhishek Chatterjee

17. Facile Site-Specific Multi-Conjugation Strategies in Recombinant Proteins Produced in Bacteria

Hannes Merten, Jonas V. Schaefer, Fabian Brandl, Uwe Zangemeister-Wittke, and Andreas Pluckthun

18. Site-Specific Photocrosslinking to Immunoglobulin G Using Photoreactive Antibody-Binding Domains

Fabiana Zappala and Andrew Tsourkas

19. SpyTag-SpyCatcher Chemistry for Protein Bioconjugation In Vitro and Protein Topology Engineering In Vivo

Xiao-Wei Wang and Wen-Bin Zhang

20. FN3 Protein Conjugates for Cancer Diagnosis and Imaging Studies

Arutselvan Natarajan and Lotfi Abou-Elkacem