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

Wagner-Rousset E; Colas O; François YN; Heinisch S; Guillarme D; Cianférani S; Beck A

doi:10.1007/978-1-4939-9929-3_12

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Journal article

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

Wagner-Rousset E Pierre Fabre Laboratories, IRPF-Centre d'Immunologie Pierre-Fabre (CIPF), Saint-Julien-en-Genevois, France.
Colas O Pierre Fabre Laboratories, IRPF-Centre d'Immunologie Pierre-Fabre (CIPF), Saint-Julien-en-Genevois, France.
François YN Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UMR 7140 (Unistra-CNRS), Université de Strasbourg, Strasbourg, France.
Heinisch S Institut des Sciences Analytiques, CNRS, UMR5280, Université de Lyon, Villeurbanne, France.
Guillarme D School of Pharmaceutical Sciences, University of Geneva, Geneva, Switzerland.
Cianférani S Laboratoire de Spectrométrie de Masse BioOrganique, IPHC UMR 7178, Université de Strasbourg, CNRS, Strasbourg, France.
Beck A Pierre Fabre Laboratories, IRPF-Centre d'Immunologie Pierre-Fabre (CIPF), Saint-Julien-en-Genevois, France. alain.beck@pierre-fabre.com.

2019-10-24

Published in:

Methods in molecular biology (Clifton, N.J.). - 2020

English 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.

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