title: Recombinant protein purification using gradient assisted simulated moving bed hydrophobic interaction chromatography. Part II: process design and experimental validation.
abstract: 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.
authors.affiliation: Otto-von-Guericke-Universität, Institut für Verfahrenstechnik, P.O. Box 4120, D-39106 Magdeburg, Germany.
authors.forename: Ludmila
authors.initials: L
authors.lastname: Gueorguieva
authors.forename: Sivakumar
authors.initials: S
authors.lastname: Palani
authors.forename: Ursula
authors.initials: U
authors.lastname: Rinas
authors.forename: Guhan
authors.initials: G
authors.lastname: Jayaraman
authors.forename: Andreas
authors.initials: A
authors.lastname: Seidel-Morgenstern
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods / Q000592:standards; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D057927:Hydrophobic and Hydrophilic Interactions; D008956:Models, Chemical; D011994:Recombinant Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results; D012685:Sepharose / Q000031:analogs & derivatives; D013300:Streptokinase / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; C052754:butylagarose; D012685:Sepharose; D013300:Streptokinase