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Abstract:
Accelerated developments of genetic engineering have increased the outbreak of biological medicines in the world market. To fulfil the regulatory requirements as well to keep the product quality, liquid chromatography plays an important role in the production process. In the last few years the Simulated Moving Bed (SMB) chromatography shows up to be an attractive alternative to batch chromatography for the production of biomolecules like Human influenza virus [1], Bone morphogenetic protein-2 (BMP) [2], monoclonal antibodies [3].
The intention of the proposed work is to develop a continuous hydrophobic interaction based SMB process for the purification of recombinant streptokinase from cell lysate. Recombinant streptokinase is a FDA approved thrombolytic protein, widely used for the congestive heart failure and peripheral vascular diseases. Recombinant Streptokinase was over expressed in E. coli and various hydrophobic interaction stationary phases (HIC) were screened for the preparative purification of recombinant streptokinase. Butyl sepharose was chosen for the experimental estimation of the adsorption isotherms of the target protein from crude cell lysate using the perturbation method [4, 5]. The determined adsorption isotherm parameters were furthermore used to design the continuous separation. The current state of the experimental work and the subsequent design of a related continuous Simulated Moving Bed process capable to isolate the target protein will be presented.
References:
[1] Kalbfuss B, Flockerzi D, Seidel-Morgenstern A, Reichl U, J. Chromatogr B, 873 (2008) 102
[2] Keßler L C, Gueorguieva L, Rinas U, Seidel-Morgenstern A, J Chromatogr. A 1176 (2007) 69.
[3] Gottschlich N, Kasche V, J. Chromatogr. A 765 (1997) 201
[4] Heuer C, Küsters E, Plattner T, Seidel-Morgenstern A, J. Chromatogr. A 827, (1998) 175
[5] Blumel C, Hugo P, Seidel-Morgenstern A, J. Chromatogr. A 865, (1999) 51
