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Expression and Purification of Recombinant Human Epidermal Growth Factor Using Fusion Partners in Escherichia coli

융합 파트너를 이용한 인간 상피세포성장인자의 재조합 대장균에서 발현과 정제 연구

  • Sung, Keehyun (Department of Chemical Engineering and Applied Chemistry, Chungnam National University) ;
  • Kim, In Ho (Department of Chemical Engineering and Applied Chemistry, Chungnam National University)
  • 성기현 (충남대학교 응용화학공학과) ;
  • 김인호 (충남대학교 응용화학공학과)
  • Received : 2018.07.04
  • Accepted : 2018.08.14
  • Published : 2018.10.01

Abstract

Human epidermal growth factor (hEGF) can stimulate the division of various cell types and has potential clinical applications. Since the protein contains three intra-molecular disulfide bonds, the high expression of active hEGF in Escherichia coli has not been well researched, We fused the hEGF gene with a small ubiquitin-related modifier gene (SUMO) by synthesizing an artificial SUMO-hEGF fusion gene that was highly expressed in E. coli (DE3) strain. The optimal expression level of the soluble fusion protein, SUMO-hEGF with IPTG (Isopropyl-${\beta}$-D-Thiogalactopyranoside), was up to 38.9% of the total cellular protein. The fusion protein was purified by Ni-NTA affinity chromatography and cleaved by a SUMO-specific protease to obtain the native hEGF, which was further purified by Ni-NTA affinity chromatography. The result of the reverse-phase HPLC showed that the purity of the recombinant cleaved hEGF was greater than 98%.

Keywords

EGF;Refolding;Expression;Purification

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