General Information
    • ISSN: 2010-0221
    • Frequency: Bimonthly
    • DOI: 10.18178/IJCEA
    • Editor-in-Chief: Prof. Dr. Shen-Ming Chen
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Editor-in-chief
Prof. Dr. Shen-Ming Chen
National Taipei University of Technology, Taiwan
 

IJCEA 2015 Vol.6(5): 306-313 ISSN: 2010-0221
DOI: 10.7763/IJCEA.2015.V6.501

Equilibrium and Kinetics of High Molecular Weight Protein Uptake in Ion Exchange Chromatography

M. Wakkel, S. Alfenore, S. Mathé, and A. Fernández
Abstract—Equilibrium and kinetic experiments have been carried out in order to study the effect of protein size in the purification of high molecular weight (HMW) proteins by IEC. The current work focused on two proteins uptake by Q HyperZ chromatographic media. Bovine Serum Albumin (BSA) and ferritin have been selected due to their close isoelectric point (4.9 and 4.4 respectively) and their different molecular weight (66.5 and 450 kDa respectively). As usually reported in the literature, equilibrium uptake for BSA has been higher than for ferritin at the same buffer conditions (pH, ionic strength), suggesting complex phenomena for ferritin uptake. Moreover, contrary to the general rule in ion-exchange processes, ferritin uptake has not been really affected by the increase of NaCl concentration. In fact, no significant change in the ferritin uptake has been quantified with the increasing of salt concentration from 0 to 100 mM. However, the kinetics of ferritin ion exchange has showed a classical behaviour, according to the homogenous diffusion model. The determination of the effective diffusion coefficient for ferritin has been possible from transient uptake at different protein initial concentrations. Thus, in spite of its large size, ferritin seems to diffuse inside the macro-porous anion-exchange particles.

Index Terms—Ion-exchange chromatography, isotherm, kinetic, protein purification, Q HyperZ.

The authors are with the Institut National des Sciences Appliquées, Toulouse, France (e-mail: wakkel@insa-toulouse.fr, alfenore@insatoulouse. fr, mathes@insa-toulouse.fr, mafernan@insa-toulouse.fr).

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Cite: M. Wakkel, S. Alfenore, S. Mathé, and A. Fernández, "Equilibrium and Kinetics of High Molecular Weight Protein Uptake in Ion Exchange Chromatography," International Journal of Chemical Engineering and Applications vol. 6, no. 5, pp. 306-313, 2015.

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