Antibody-Drug Conjugates: Methods and Protocols

Front Matter ....Pages i-xi
 An Overview of the Current ADC Discovery Landscape  (L. Nathan Tumey)....Pages 1-22
 Pushing the Envelope: Advancement of ADCs Outside of Oncology  (Michael J. McPherson, Adrian D. Hobson)....Pages 23-36
 Conjugations to Endogenous Cysteine Residues  (Durgesh V. Nadkarni)....Pages 37-49
 Site-Specific Conjugation to Cys-Engineered THIOMAB™ Antibodies  (Pragya Adhikari, Neelie Zacharias, Rachana Ohri, Jack Sadowsky)....Pages 51-69
 Transglutaminase-Mediated Conjugations  (Yasuaki Anami, Kyoji Tsuchikama)....Pages 71-82
 Click Chemistry Conjugations  (Tak Ian Chio, Susan L. Bane)....Pages 83-97
 Utilizing Solid-Phase to Enable High-Throughput, Site-Specific Conjugation and Dual-Labeled Antibody and Fab Conjugates  (Sujiet Puthenveetil)....Pages 99-112
 Bridged Cysteine Conjugations  (Matthew Bird, Joao Nunes, Mark Frigerio)....Pages 113-129
 Antibody Conjugations via Glycosyl Remodeling  (Hanna Toftevall, Helén Nyhlén, Fredrik Olsson, Jonathan Sjögren)....Pages 131-145
 ADC Analysis by Hydrophobic Interaction Chromatography  (Ryan Fleming)....Pages 147-161
 Two-Dimensional Liquid Chromatography Coupled to High-Resolution Mass Spectrometry for the Analysis of ADCs  (Soraya Chapel, Florent Rouvière, Morgan Sarrut, Sabine Heinisch)....Pages 163-185
 Drug Loading and Distribution of ADCs After Reduction or IdeS Digestion and Reduction  (Elsa Wagner-Rousset, Olivier Colas, Yannis-Nicolas François, Sabine Heinisch, Davy Guillarme, Sarah Cianférani et al.)....Pages 187-195
 Analysis of ADCs by Native Mass Spectrometry  (Oscar Hernandez-Alba, Anthony Ehkirch, Alain Beck, Sarah Cianférani)....Pages 197-211
 High-Resolution Characterization of ADCs by Orbitrap LCMS  (Jintang He, Surinder Kaur, Keyang Xu)....Pages 213-219
 Conjugation Site Analysis by MS/MS Protein Sequencing  (Linjie Han, Yanqun Zhao, Qunying Zhang)....Pages 221-233
 Conjugation Site Analysis of Lysine-Conjugated ADCs  (Hua Sang, Ning Wan, Gaoyuan Lu, Yang Tian, Guangji Wang, Hui Ye)....Pages 235-250
 Characterization of ADCs by Capillary Electrophoresis  (Wenjing Ning, Yanqun Zhao)....Pages 251-262
 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, Yannis-Nicolas François)....Pages 263-272
 Purification of ADCs by Hydrophobic Interaction Chromatography  (Calvin L. Becker, Robert J. Duffy, Jorge Gandarilla, Steven M. Richter)....Pages 273-290
 Detection and Removal of Small Molecule and Endotoxin Contaminants in ADC Preparations  (Jeffrey Casavant, Anokha S. Ratnayake, Sujiet Puthenveetil, L. Nathan Tumey)....Pages 291-299
 Physical Stability Studies of Antibody-Drug Conjugates (ADCs) Under Stressed Conditions  (Yilma T. Adem)....Pages 301-311
 Biophysical Methods for Characterization of Antibody-Drug Conjugates  (Vamsi Krishna Mudhivarthi, Jianxin Guo)....Pages 313-327
 Determination of ADC Cytotoxicity in Immortalized Human Cell Lines  (Shengjia Wu, Dhaval K. Shah)....Pages 329-340
 LC/MS Methods for Studying Lysosomal ADC Catabolism  (Andrew J. Bessire, Chakrapani Subramanyam)....Pages 341-351
 Assessing ADC Plasma Stability by LC-MS Methods  (Cong Wei)....Pages 353-359
 Determination of ADC Concentration by Ligand-Binding Assays  (Hsuan-Ping Chang, Dhaval K. Shah)....Pages 361-369
Back Matter ....Pages 371-373