Hydrophobic interaction chromatography of proteins III. Transport and kinetic parameters in isocratic elution

Journal of Chromatography. a
Brian C S To, Abraham M Lenhoff

Abstract

The effective pore diffusivities, D(e), of five model proteins (ribonuclease A, lysozyme, alpha-lactalbumin, ovalbumin, and BSA) in eight commercial phenyl hydrophobic interaction chromatography (HIC) media were determined by analyzing the plate height data from isocratic elution using the first two moments of the general linear rate model. The adsorbents represent a diverse set of HIC media that are widely used for protein purification. The estimated pore diffusivities were used to calculate the elution profiles of proteins in these adsorbents and were compared with the elution profiles obtained experimentally. High protein loading and sample protein concentration led to the underestimation of the pore diffusivity by the linear rate model. Comparisons between the calculated and the experimental profiles suggest that the pore diffusivities obtained from the linear rate model are generally accurate for proteins with low structural flexibility but not for more flexible ones, presumably because conformational change effects contribute significantly to the overall HETP. The general linear rate model was modified to account for the protein folding/unfolding kinetics, and parameter values could be estimated by fitting the experimenta...Continue Reading

References

Oct 21, 1986·Biochemistry·K Gekko, Y Hasegawa
Jan 1, 1993·Biopolymers·D P Kharakoz, A P Sarvazyan
Feb 16, 2000·Journal of Chromatography. a·R K Lewus, G Carta
Jul 21, 2006·Journal of Chromatography. a·J F LangfordA M Lenhoff
Nov 23, 2006·Journal of Chromatography. a·Emmerich HaimerAlois Jungbauer

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Citations

Jun 15, 2011·Journal of Chromatography. a·Fabrice Gritti, Georges Guiochon
Nov 1, 2012·Journal of Separation Science·Rainer Hahn
Dec 31, 2010·Biotechnology and Bioengineering·Guo-Zhen WangYan Sun
Nov 10, 2012·Biotechnology Journal·Kristian KallbergLeif Bulow
Apr 7, 2014·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Dong-Xia HaoZhi-Guo Su

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