Book chapter
English

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

Published inL. Nathan Tumey (Ed.), Antibody-Drug Conjugates Methods and Protocols
PublisherNew York, NY : Springer US
Publication date2020
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 vedotin
  • DAR
  • DLD
  • Fabricator
  • HIC
  • IdeS
  • Mass spectrometry
Research groups
Citation (ISO format)
WAGNER-ROUSSET, Elsa et al. Drug Loading and Distribution of ADCs After Reduction or IdeS Digestion and Reduction. In: Antibody-Drug Conjugates Methods and Protocols. L. Nathan Tumey (Ed.). New York, NY : Springer US, 2020. doi: 10.1007/978-1-4939-9929-3_12
Main files (1)
Book chapter (Published version)
accessLevelRestricted
Identifiers
ISBN978-1-4939-9928-6
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Technical informations

Creation12/11/2019 11:45:00
First validation12/11/2019 11:45:00
Update15/03/2023 18:22:54
Status update15/03/2023 18:22:53
Last indexation31/10/2024 16:54:28
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