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Article Dans Une Revue Electrophoresis Année : 2018

High-resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method

Alexandre Goyon
  • Fonction : Auteur
Yannis Francois
Olivier Colas
  • Fonction : Auteur
Alain Beck
  • Fonction : Auteur
Jean Luc Veuthey
  • Fonction : Auteur
Davy Guillarme
  • Fonction : Auteur

Résumé

The determination of mAb critical quality attributes (CQA) is crucial for their successful application in health diseases. A generic CZE method was developed for the high-resolution separation of various mAb charge variants, which are often recognized as important CQA. A dynamic coating of the capillary was obtained with polyethylene oxide (PEO), whereas Bis-Tris allowed the analysis of mAbs under native conditions at pH 7.0. The effect of PEO and Bis-Tris concentrations, as well as the nature of the acidic counter ion on the method performance were systematically studied. The %RSD on migration times was below 5 % on three different CE instruments using the optimized method. Additional charge variants (in particular acidic variants) were resolved for 10 out of 17 mAbs compared to a reference CZE approach involving the use of ε-amino-caproic acid (EACA), triethylenetetramine (TETA), and hydroxypropylmethyl cellulose (HPMC). The amount of basic and acidic charge variants of 17 Food and Drug Administration (FDA) approved mAbs covering a wide range of physico-chemical properties, e.g. pI between 8.0 and 9.4 and different hydrophobicity, were mainly comprised between 5 – 15 % and 15 – 30%, respectively. It is noteworthy that applications for the quality control in hospitals as well as for the combination of the immune checkpoint inhibitors nivolumab and ipilimumab were presented.
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Dates et versions

hal-03039576 , version 1 (03-12-2020)

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Citer

Alexandre Goyon, Yannis Francois, Olivier Colas, Alain Beck, Jean Luc Veuthey, et al.. High-resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method. Electrophoresis, 2018, 39 (16), pp.2083-2090. ⟨10.1002/elps.201800131⟩. ⟨hal-03039576⟩
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