KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Comparative Analysis of Dissolution and Refolding Processes for Inclusion Body Protein Renaturation

내포체 단백질 재생을 위한 용해 및 재접힘공정의 비교분석

  • 김창성 (한양대학교 화학공학과 생물공정연구실) ;
  • 김윤하 (쌍용정유 기술개발팀) ;
  • 이은규 (한양대학교 화학공학과 생물공정연구실)
  • Published : 1998.04.01
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Abstract

Using rlFN-$\alpha$ and rhGH as the model proteins, the refolding performances of the published processes were evaluated and compared. Key engineering parameters such as the type of denaturant and this concentration, protein concentration in the refolding buffer, and pH and ionic strength of the buffer were experimentally investigated. Furthermore, the role of a co-solvent of surfactant type in aggregation reduction was also studied. Of the denaturants tested (8M urea, 6M guanidine HCI, 0.5% SDS), SDS at alkaline pH (9.5) and ambient temperature gave the highest recovery yield. The SDS process was effective in the refolding of observed where dissolution proceeded better under lower strength (10 mM) but aggregation was suppressed under higher strength (>50 mM.) When PEG-4000 and/or Tween were added as co-solvent or refolding-enhancing additive, 1.6-2 times higher yield was realized. The‘masking’of the hyrophobic patches located on the surface of the protein with the surfactant molecules was believed to be responsible for the considerable reduction in aggregation during refolding.

Keywords

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