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 FrançoisEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2078)


Capillary electrophoresis-mass spectrometry (CE-MS) enables the characterization of the primary structure of ADCs. An analytical method based on a derived bottom-up proteomic workflow is designed to provide detailed information about the amino acid sequence, the glycosylation profiling, and the location on the peptide backbone of the conjugated drugs. Here we describe the experimental protocol applied on the characterization of cysteine-linked brentuximab vedotin (Adcetris®).

Key words

Antibody-drug conjugate Brentuximab vedotin Cytotoxic drug Capillary electrophoresis–mass spectrometry Structural characterization Drug-loaded peptide 



The authors would like to thank Sciex separations Inc. for lending a CESI8000 system and a 5600 TripleTOF. They would like to thank Bruker Daltonics for lending a Maxis 4G. They would like also to express their gratitude to Dr. E. Wagner-Rousset, M. C. Janin-Bussat, and O. Colas (Centre d’Immunologie Pierre Fabre, St Julien en Genevois, France) for helpful discussions. This work was supported by the CNRS (UMR 7140) and the University of Strasbourg.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Josiane Saadé
    • 1
  • Rabah Gahoual
    • 2
  • Alain Beck
    • 3
  • Emmanuelle Leize-Wagner
    • 1
  • Yannis-Nicolas François
    • 1
    Email author
  1. 1.Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), UMR 7140 (Unistra-CNRS)Université de StrasbourgStrasbourgFrance
  2. 2.Laboratoire Vecteurs Pour l’Imagerie Moléculaire et le Ciblage Thérapeutique (VICT), Faculté de PharmacieUniversité Paris DescartesParisFrance
  3. 3.Pierre Fabre LaboratoriesIRPF—Centre d’Immunologie Pierre Fabre (CIPF)Saint-Julien-en-GenevoisFrance

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