1. An Overview of the Current ADC Discovery Landscape L. Nathan Tumey
2. Pushing the Envelope: Advancement of ADCs Outside of Oncology Michael J. McPherson and Adrian D. Hobson
3. Conjugations to Endogenous Cysteine Residues Durgesh V. Nadkarni
4. Site-Specific Conjugation to Cys-Engineered THIOMAB™ Antibodies Pragya Adhikari, Neelie Zacharias, Rachana Ohri, and Jack Sadowsky
5. Transglutaminase-Mediated Conjugations Yasuaki Anami and Kyoji Tsuchikama
6. Click Chemistry Conjugations Tak Ian Chio and Susan L. Bane
7. Utilizing Solid-Phase to Enable High-Throughput, Site-Specific Conjugation and Dual Labeled Antibody and Fab Conjugates Sujiet Puthenveetil
8. Bridged Cysteine Conjugations Mark Frigerio, Joao Nunes, and Matthew Bird
9. Antibody Conjugations via Glycosyl Remodeling Hanna Toftevall, Helén Nyhlén, Fredrik Olsson, and Jonathan Sjögren
10. ADC Analysis by Hydrophobic Interaction Chromatography Ryan Fleming
11. Two-Dimensional Liquid Chromatography Coupled to High Resolution Mass Spectrometry for the Analysis of ADCs Soraya Chapel, Florent Rouvière, Morgan Sarrut, and Sabine Heinisch
12. Drug Loading and Distribution of ADCs after Reduction or IdeS Digestion and Reduction Elsa Wagner-Rousset, Olivier Colas, Yannis François, Sabine Heinisch, Davy Guillarme, Sarah Cianférani, and Alain Beck
13. Analysis of ADCs by Native Mass Spectrometry Oscar Hernandez-Alba, Anthony Ehkirch, Alain Beck, and Sarah Cianférani
14. High Resolution Characterization of ADCs by Orbitrap LCMS Jintang He, Surinder Kaur, and Keyang Xu
15. Conjugation Site Analysis by MS/MS Protein Sequencing Linjie Han, Yanqun Zhao, and Qunying Zhang
16. Conjugation Site Analysis of Lysine-Conjugated ADCs Hua Sang, Ning Wan, Gaoyuan Lu, Yang Tian, Guangji Wang, and Hui Ye
17. Characterization of ADCs by Capillary Electrophoresis Wenjing Ning and Yanqun Zhao
18. Characterization of the Primary Structure of Cysteine-Linked Antibody-Drug Conjugates using Capillary Electrophoresis with Mass Spectrometry Josiane Saadé, Rabah Gahoual, Alain Beck, Emmanuelle Leize-Wagner, and Yannis-Nicolas François
19. Purification of ADCs by Hydrophobic Interaction Chromatography Calvin L. Becker, Robert J. Duffy, Jorge Gandarilla, and Steven M. Richter
20. Detection and Removal of Small-Molecule and Endotoxin Contaminants in ADC Preparations Jeffery Casavant, Anokha S. Ratnayake, Sujiet Puthenveetil, and L. Nathan Tumey
21. Physical Stability Studies of Antibody Drug Conjugates (ADCs) Under Stressed Conditions Yilma T. Adem
22. Biophysical Methods for Characterization of Antibody Drug Conjugates Vamsi Krishna Mudhivarthi and Jianxin Guo
23. Determination of ADC Cytotoxicity in Immortalized Human Cell Lines Shengjia Wu and Dhaval K. Shah
24. LC/MS Methods for Studying Lysosomal ADC Catabolism Andrew J. Bessire and Chakrapani Subramanyam
25. Assessing ADC Plasma Stability by LC/MS Methods Cong Wei
26. Determination of ADC Concentration by Ligand-Binding Assays Hsuan-Ping Chang and Dhaval K. Shah
This volume looks at key methodologies that are commonly used across antibody drug conjugates (ADCs) programs. The chapters in this book cover topics such as conjugations to endogenous cysteine residues; click chemistry conjugations; antibody conjugations via glycosyl remodeling; analysis of ADCs by native mass spectrometry; characterization of ADCs by capillary electrophoresis; LC/MS methods for studying lysosomal ADC catabolism; and determination of ADC concentration by ligand-binding assays. 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 practical, Antibody-Drug Conjugates: Methods and Protocols is a valuable resource that aims to lower the “activation barrier” when undertaking a new discipline, and provides a “toolbox” for the next generation of ADC scientists.