Mechanistic Modeling of Reversed-Phase Chromatography of Insulins with Potassium Chloride and Ethanol as Mobile-Phase Modulators

ACS Omega
Karolina ArkellBernt Nilsson

Abstract

The purpose of this study was to investigate the adsorption mechanism in reversed-phase chromatography (RPC) of proteins and to develop a model for the effect of dual mobile phase modulators-a salt and an organic solvent-on this process. Two different adsorption mechanisms were considered: (1) pure association of a protein molecule and one or more ligands and (2) displacement of the organic modulator, with which the adsorbent is saturated, by the protein upon association with one or more ligands. One model was then derived from each of the two considered mechanisms, combining thermodynamic theories on salting-in, RPC, and the solubility of proteins. The model was then applied to chromatographic data from an earlier report as well as supplementary data for solubility and vapor-liquid equilibria, and case-specific simplifications were made. We found that an adaptation of Kirkwood's electrostatic theories to hydrophobic interaction chromatography describes the observed effect of KCl well. Combining chromatographic and solubility data for one of the insulins, we concluded that the variation in the activity coefficient of the insulin with respect to the concentration of ethanol alone cannot describe its effect on retention. Conseque...Continue Reading

References

May 1, 1984·Physics in Medicine and Biology·A A LaogunE H Grant
Jan 29, 1999·Journal of Chromatography. a·A Vailaya, C Horváth
Jan 1, 1959·Advances in Protein Chemistry·W KAUZMANN
Feb 11, 2005·Journal of Chromatography. a·Niklas JakobssonBernt Nilsson
Mar 24, 2005·Journal of Chromatography. a·Alois Jungbauer
Oct 8, 2008·Journal of Chromatography. a·P Nikitas, A Pappa-Louisi
Jul 26, 2014·Physical Chemistry Chemical Physics : PCCP·Rico F TaborDerek Y C Chan

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Citations

Apr 3, 2019·Biotechnology Progress·Patricia RochCarl-Fredrik Mandenius
Apr 28, 2018·Biotechnology Letters·Anurag S RathoreNikhil Kateja
Jun 10, 2020·Annual Review of Chemical and Biomolecular Engineering·Vijesh Kumar, Abraham M Lenhoff

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Methods Mentioned

BETA
reversed-phase chromatography

Software Mentioned

MATLAB

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