Scientific article
Open access

Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers

Published inInternational journal of molecular sciences, vol. 24, no. 23, p. 16623
Publication date2023-11-22
First online date2023-11-22

Charge heterogeneity among therapeutic monoclonal antibodies (mAbs) is considered an important critical quality attribute and requires careful characterization to ensure safe and efficacious drug products. The charge heterogeneity among mAbs is the result of chemical and enzymatic post-translational modifications and leads to the formation of acidic and basic variants that can be characterized using cation exchange chromatography (CEX). Recently, the use of mass spectrometry-compatible salt-mediated pH gradients has gained increased attention to elute the proteins from the charged stationary phase material. However, with the increasing antibody product complexity, more and more selectivity is required. Therefore, in this study, we set out to improve the selectivity by using a solvent-enriched mobile phase composition for the analysis of a variety of mAbs and bispecific antibody products. It was found that the addition of the solvents to the mobile phase appeared to modify the hydrate shell surrounding the protein and alter the retention behavior of the studied proteins. Therefore, this work demonstrates that the use of solvent-enriched mobile phase composition could be an attractive additional method parameter during method development in CEX.

  • Fab fragment
  • Bispecific antibody
  • Ion exchange chromatography
  • Monoclonal antibody
  • Organic modifier
  • Salt gradient
  • Salt-mediated pH gradient
  • Roche Diagnostics GmbH -
Citation (ISO format)
DUIVELSHOF, Bastiaan et al. Enhancing Selectivity of Protein Biopharmaceuticals in Ion Exchange Chromatography through Addition of Organic Modifiers. In: International journal of molecular sciences, 2023, vol. 24, n° 23, p. 16623. doi: 10.3390/ijms242316623
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Article (Published version)
ISSN of the journal1422-0067

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