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http://dx.doi.org/10.9713/kcer.2018.56.5.711

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

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)
Publication Information
Korean Chemical Engineering Research / v.56, no.5, 2018 , pp. 711-717 More about this Journal
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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Times Cited By KSCI : 1  (Citation Analysis)
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