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Title

Drug Loading and Distribution of ADCs After Reduction or IdeS Digestion and Reduction

Authors
Wagner-Rousset, Elsa
Colas, Olivier
François, Yannis-Nicolas
Heinisch, Sabine
Cianférani, Sarah
Beck, Alain
Published in L. Nathan Tumey. Antibody-Drug Conjugates Methods and Protocols. New York, NY: Springer US. 2020
Abstract High-resolution native mass spectrometry (MS) provides accurate mass measurements (within 30 ppm) of intact ADCs and can also yield drug load distribution (DLD) and average drug to antibody ratio (DAR) in parallel with hydrophobic interaction chromatography (HIC). Native MS is furthermore unique in its ability to simultaneously detect covalent and noncovalent species in a mixture and for HIC peak identity assessment offline or online. As an orthogonal method described in this chapter, LC-MS following ADC reduction or IdeS (Fabricator) digestion and reduction can also be used to measure the DLD of light chain and Fd fragments for hinge native cysteine residues such as brentuximab vedotin. Both methods allow also the measurement of average DAR for both monomeric and multimeric species. In addition, the Fc fragments can be analyzed in the same run, providing a complete glycoprofile and the demonstration or absence of additional conjugation of this subdomain involved in FcRn and Fc-gammaR binding.
Keywords Brentuximab vedotinDARDLDFabricatorHICIdeSMass spectrometry
Identifiers
PMID: 31643057
ISBN: 978-1-4939-9928-6
Full text
Structures
Research group Sciences analytiques
Citation
(ISO format)
WAGNER-ROUSSET, Elsa et al. Drug Loading and Distribution of ADCs After Reduction or IdeS Digestion and Reduction. In: L. Nathan Tumey (Ed.). Antibody-Drug Conjugates Methods and Protocols. New York, NY : Springer US, 2020. doi: 10.1007/978-1-4939-9929-3_12 https://archive-ouverte.unige.ch/unige:126333

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Deposited on : 2019-11-15

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