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Tuning selectivity in cation-exchange chromatography applied for monoclonal antibody separations, part 1: Alternative mobile phases and fine tuning of the separation.
Journal article

Tuning selectivity in cation-exchange chromatography applied for monoclonal antibody separations, part 1: Alternative mobile phases and fine tuning of the separation.

  • Farsang E Department of Analytical Chemistry, University of Pannonia, Egyetem u. 10, 8200 Veszprém, Hungary.
  • Murisier A School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, CMU - Rue Michel Servet, 1, 1211 Geneva 4, Switzerland.
  • Horváth K Department of Analytical Chemistry, University of Pannonia, Egyetem u. 10, 8200 Veszprém, Hungary.
  • Beck A Center of Immunology Pierre Fabre, 5 Avenue Napoléon III, BP 60497, 74160 Saint-Julien-en-Genevois, France.
  • Kormány R Egis Pharmaceuticals Plc., Keresztúri út 30-38, Budapest 1106, Hungary.
  • Guillarme D School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, CMU - Rue Michel Servet, 1, 1211 Geneva 4, Switzerland.
  • Fekete S School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, CMU - Rue Michel Servet, 1, 1211 Geneva 4, Switzerland. Electronic address: szabolcs.fekete@unige.ch.
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  • 2019-02-27
Published in:
  • Journal of pharmaceutical and biomedical analysis. - 2019
Cation-exchange chromatography
Method development
Monoclonal antibody
Salt gradient
Salt-mediated pH gradient
pH gradient
Antibodies, Monoclonal
Buffers
Cations
Chromatography, Ion Exchange
Hydrogen-Ion Concentration
Sodium Chloride
Software
English Cation exchange chromatography (CEX) of therapeutic monoclonal antibodies is generally performed with either salt gradient (MES buffer + NaCl) or using commercial pH gradient buffer. The goal of this study was to find out some alternative buffer systems for CEX separation of mAbs, which may offer alternative selectivity, while maintaining similar peak shapes. Among the new buffers that were tested, (N-morpholino)ethanesulfonic acid (MES) / 1,3-diamino-2-propanol (DAP), and citric acid / 2-(cyclohexylamino)ethanesulfonic acid (CHES) systems were particularly promising, especially when combining them with a moderate salt gradient of NaCl. This two buffer system provides an equivalent or slightly better separation than the standard, mobile phases for therapeutic mAbs. It was also demonstrated that working with salt-mediated pH gradients, allows to extend the possibilities in method development, since the concentration of salt in the mobile phase has a significant impact on selectivity. Using HPLC modeling software (Drylab), it was possible to successfully develop CEX methods for authentic mAb samples within only 6 h, by optimizing the gradient steepness and salt concentration in the B eluent.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/46730
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