Abstract

Introduction: Wide applications in research, clinical and cosmetic industry of human epidermal growth factor (hEGF) has made it as a research interest target. Its production in different expression systems have shown several limitations. Materials and Methods: Recombinant expression of hEGF in Escherichia coli is always accompanied with inclusion body formation. In order to evaluate a chromatogram independent purification approach for recombinant hEGF production in soluble form, the hEGF gene was fused to an elastin-like protein (ELP) and was expressed in E. coli BL21 (DE3) using pET26b expression vector for secreting the product into periplasmic space. Results: Periplasmic protein content analysis confirmed that the recombinant protein is secreted into periplasm. Purification process was done by using 0.4M ammonium sulfate in two cycles of inverse transition cycling (ITC). After two cycles of purification, the purity reached more than 95. Western blotting analysis with monoclonal anti-EGF anti body have confirmed the accuracy of EGF. The biological activity of the purified protein was investigated on the NIH-3T3 cell line and results indicated that EGF-induced proliferation in treated cells. Conclusions: Results revealed that the periplasmic expression is a suitable approach to produce soluble recombinant hEGF. By using of ELP fused to EGF, the purification process was stablished without applying chromatography which will result in decreasing final costs. This study introduced a new economic and efficient approach for the production and purification of recombinant hEGF. © 2021 The Author(s).