Determination of the Adsorption Isotherm Parameters from Total Protein Mixture

As a versatile tool, liquid chromatography has become an inevitable part of protein separation and purification processes. In the last two decades, chromatography has been very successfully applied for the purification of various target proteins from the mixtures of complex host cell proteins. In the last years, the continuous Simulated Moving Bed (SMB) process has started finding its applications for protein purification1. For the design of such SMB processes, the accurate estimation of adsorption isotherm parameters of the target components is very important. In the literature there are several methods describing the experimental determination of equilibrium isotherms of pure compounds and of complex mixtures2. Commonly the pure target protein of interest is not commercially available at an affordable price. In such cases an efficient direct estimation of the adsorption isotherms from the protein mixtures (fermentation broths or crude cell lysate) will decrease the process development time. Recombinant streptokinase is a FDA approved thrombolytic protein widely used for the treatment of congestive heart failure and peripheral vascular diseases. The purpose of this study is to explore the potential and applicability of the perturbation method3,4 for determining the adsorption isotherm parameters for recombinant streptokinase from crude cell lysate (multi-component mixture). The current state of the corresponding experimental work on recombinant streptokinase purification and the subsequent design of a related continuous SMB process capable to isolate the target protein will be presented. References: (1) Nicoud R M Subramanian G(ed.) in Bioseparation and Bioprocessing A hand book Vol I Wiley-VCH, Germany, 1998. (2) Seidel-Morgenstern A, J. Chromatogr. A 1037,(2004) 255. (3) Heuer C, Küsters E, Plattner T, Seidel-Morgenstern A, J. Chromatogr. A 827,(1998) 175. (4) Blumel C, Hugo P, Seidel-Morgenstern A, J. Chromatogr. A 865,(1999) 51