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.
Reihe
Sprache
Verlagsort
Verlagsgruppe
Illustrationen
109
29 s/w Abbildungen, 109 farbige Abbildungen
XI, 373 p. 138 illus., 109 illus. in color.
Dateigröße
ISBN-13
978-1-4939-9929-3 (9781493999293)
DOI
10.1007/978-1-4939-9929-3
Schweitzer Klassifikation
An Overview of the Current ADC Discovery Landscape.- Pushing the Envelope: Advancement of ADCs Outside of Oncology.- Conjugations to Endogenous Cysteine Residues.- Site-Specific Conjugation to Cys-Engineered THIOMABT Antibodies.- Transglutaminase-Mediated Conjugations.- Click Chemistry Conjugations.- Utilizing Solid-Phase to Enable High-Throughput, Site-Specific Conjugation and Dual Labeled Antibody and Fab Conjugates.- Bridged Cysteine Conjugations.- Antibody Conjugations via Glycosyl Remodeling.- ADC Analysis by Hydrophobic Interaction Chromatography.- Two-Dimensional Liquid Chromatography Coupled to High Resolution Mass Spectrometry for the Analysis of ADCs.- Drug Loading and Distribution of ADCs after Reduction or IdeS Digestion and Reduction.- Analysis of ADCs by Native Mass Spectrometry.- High Resolution Characterization of ADCs by Orbitrap LCMS.- Conjugation Site Analysis by MS/MS Protein Sequencing.- Conjugation Site Analysis of Lysine-Conjugated ADCs.- Characterization of ADCs by Capillary Electrophoresis.- Characterization of the Primary Structure of Cysteine-Linked Antibody-Drug Conjugates using Capillary Electrophoresis with Mass Spectrometry.- Purification of ADCs by Hydrophobic Interaction Chromatography.- Detection and Removal of Small-Molecule and Endotoxin Contaminants in ADC Preparations.- Physical Stability Studies of Antibody Drug Conjugates (ADCs) Under Stressed Conditions.- Biophysical Methods for Characterization of Antibody Drug Conjugates.- Determination of ADC Cytotoxicity in Immortalized Human Cell Lines.- LC/MS Methods for Studying Lysosomal ADC Catabolism.- Assessing ADC Plasma Stability by LC/MS Methods.- Determination of ADC Concentration by Ligand-Binding Assays.
