TY - JOUR

T1 - Recombinant protein purification using gradient assisted simulated moving bed hydrophobic interaction chromatography. Part II

T2 - Process design and experimental validation

AU - Gueorguieva, Ludmila

AU - Palani, Sivakumar

AU - Rinas, Ursula

AU - Jayaraman, Guhan

AU - Seidel-Morgenstern, Andreas

N1 - Funding Information: We acknowledge the support of the Bioprocess Engineering Group at the MPI and the Leibniz Institute for Neurobiology (both in Magdeburg). S.P. acknowledges the financial support of Deutscher Akademischer Austauschdienst (DAAD) and the travel grant to the “HPLC 2009” from GDCh . The support of Deutsche Forschungsgemeinschaft via “ SFB 578 ” is also gratefully acknowledged.

PY - 2011/9/16

Y1 - 2011/9/16

N2 - In the first part of this work adsorption isotherm parameters were acquired to describe the migration of recombinant streptokinase in Butyl Sepharose columns at different salt concentrations. Based on these results, a simulated moving bed (SMB) chromatographic process was designed and realised, which exploits a two-step salt gradient and allows the continuous separation of streptokinase from contaminants present in a clarified Escherichia coli cell lysate solution. This second part describes the design of the three-zone open-loop gradient SMB process applying both equilibrium theory and an equilibrium stage model and presents results of a series of experiments aiming to obtain pure streptokinase. Moreover, the potential of the SMB process and the design approach are evaluated.

AB - In the first part of this work adsorption isotherm parameters were acquired to describe the migration of recombinant streptokinase in Butyl Sepharose columns at different salt concentrations. Based on these results, a simulated moving bed (SMB) chromatographic process was designed and realised, which exploits a two-step salt gradient and allows the continuous separation of streptokinase from contaminants present in a clarified Escherichia coli cell lysate solution. This second part describes the design of the three-zone open-loop gradient SMB process applying both equilibrium theory and an equilibrium stage model and presents results of a series of experiments aiming to obtain pure streptokinase. Moreover, the potential of the SMB process and the design approach are evaluated.

KW - Hydrophobic interaction chromatography

KW - Streptokinase

KW - Two-step gradient open-loop simulated moving bed chromatography

UR - http://www.scopus.com/inward/record.url?scp=80051870442&partnerID=8YFLogxK

U2 - 10.1016/j.chroma.2011.07.008

DO - 10.1016/j.chroma.2011.07.008

M3 - Article

C2 - 21824621

AN - SCOPUS:80051870442

VL - 1218

SP - 6402

EP - 6411

JO - Journal of Chromatography A

JF - Journal of Chromatography A

SN - 0021-9673

IS - 37

ER -