Main description:

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.

Contents:

Preface...
Table of Contents...
Contributing Authors...

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 (TM) 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, Helen Nyhlen, Fredrik Olsson, and Jonathan Sjoegren

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 Rouviere, 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 Francois, Sabine Heinisch, Davy Guillarme, Sarah Cianferani, and Alain Beck

13. Analysis of ADCs by Native Mass Spectrometry
Oscar Hernandez-Alba, Anthony Ehkirch, Alain Beck, and Sarah Cianferani

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 Saade, Rabah Gahoual, Alain Beck, Emmanuelle Leize-Wagner, and Yannis-Nicolas Francois

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