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Perturbation method; Adsorption isotherm parameters; Simulated moving bed; Hydrophobic interaction chromatography; Recombinant streptokinase
Abstract:
The design of gradient simulated moving bed (SMB) chromatographic processes requires after selection of the chromatographic system the determination of adsorption isotherm parameters in the relevant range of mobile phase conditions. The determination of these parameters can be quite difficult for recombinant target proteins present in complex protein mixtures. The first part of this work includes the estimation of adsorption isotherm parameters for streptokinase and a lumped impurity fraction present in an E.coli cell lysate for a hydrophobic interaction chromatography (HIC) matrix. Perturbation experiments were carried out using a Butyl Sepharose matrix with purified recombinant protein on buffer equilibrated columns as well as with crude cell lysate saturated columns. The Henry constants estimated for streptokinase were found to exhibit in a wide range a linear dependence on the salt concentration in the mobile phase. These parameters were applied in subsequent investigations to design a simulated moving bed (SMB) process capable to purify in a continuous manner recombinant streptokinase from the E.coli cell lysate.
Copyright © 2011 Published by Elsevier B.V. [accessed September 1st 2011]
