Recombinant protein purification using gradient assisted simulated moving bed hydrophobic interaction chromatography. Part II: process design and experimental validation

J Chromatogr A. 2011 Sep 16;1218(37):6402-11. doi: 10.1016/j.chroma.2011.07.008. Epub 2011 Jul 18.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Liquid* / methods
  • Chromatography, Liquid* / standards
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli
  • Hydrophobic and Hydrophilic Interactions
  • Models, Chemical*
  • Recombinant Proteins / isolation & purification*
  • Reproducibility of Results
  • Sepharose / analogs & derivatives
  • Streptokinase / isolation & purification

Substances

  • Recombinant Proteins
  • butylagarose
  • Sepharose
  • Streptokinase