Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Refolding and Purification of Recombinant Human $Interferon-\gamma$ Expressed as Inclusion Bodies in Escherichia coli Using Size Exclusion Chromatography

  • Guan Yi-Xin (Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Pan Hai-Xue (Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Gao Yong-Gui (Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Yao Shan-Jing (Department of Chemical and Biochemical Engineering, Zhejiang University) ;
  • Cho Man-Gi (Advanced Science and Technology Research Center, Graduate School of Biotechnology, Dongseo University)
  • Published : 2005.03.01
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Abstract

A size exclusion chromatography (SEC) process, in the presence of denaturant in the refolding buffer was developed to refold recombinant human $interferon-\gamma$ ($rhIFN-\gamma$) at a high concentration. The $rhlFN-\gamma$ was overexpressed in E. coli resulting in the formation of inactive inclusion bodies (IBs). The IBs were first solubilized in 8 M urea as the denaturant, and then the refolding process performed by decreasing the urea concentration on the SEC column to suppress protein aggregation. The effects of the urea concentration, protein loading mode and column height during the refolding step were investigated. The combination of the buffer-exchange effect of SEC and a moderate urea concentration in the refolding buffer resulted in an efficient route for producing correctly folded $rhIFN-\gamma$, with protein recovery of $67.1\%$ and specific activity up to $1.2\times10^7\;IU/mg$.

Keywords

References

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