Expression optimization, purification and in vitro characterization of human epidermal growth factor produced in Nicotiana benthamiana

Oranicha Hanittinan; Yamin Oo; Chatchai Chaotham; Kaewta Rattanapisit; Balamurugan Shanmugaraj; Waranyoo Phoolcharoen

doi:10.1016/j.btre.2020.e00524

Expression optimization, purification and in vitro characterization of human epidermal growth factor produced in Nicotiana benthamiana

Biotechnol Rep (Amst). 2020 Sep 5:28:e00524. doi: 10.1016/j.btre.2020.e00524. eCollection 2020 Dec.

Authors

Oranicha Hanittinan¹, Yamin Oo², Chatchai Chaotham^{2

3}, Kaewta Rattanapisit^{1

4}, Balamurugan Shanmugaraj^{1

4}, Waranyoo Phoolcharoen^{1

4}

Affiliations

¹ Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
² Department of Biochemistry and Microbiology, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
³ Cell-based Drug and Health Products Development Research Unit, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
⁴ Research Unit for Plant-produced Pharmaceuticals, Chulalongkorn University, Bangkok, Thailand.

Abstract

Human epidermal growth factor (hEGF) has gained clinical importance due to its ability to promote wound healing. Due to its commercial applications and high market demand, recombinant EGF has been produced in several forms. Currently, plant expression system is considered as potential alternative for low-cost recombinant protein production. Hence, this study focused on improving the production of hEGF in plants by effective gene construct design and optimizing the Agrobacterium culture conditions for high protein production. In this context, hEGF gene was cloned into plant geminiviral expression vector pBYR2e and transformed in to N. benthamiana leaves via., agroinfiltration. The recombinant hEGF was purified from the plant crude extracts by single-step affinity chromatography. Furthermore, the plant-produced hEGF has shown to promote cell migration comparable to commercial hEGF in HaCaT cells in vitro. These results indicated the potential of plant expression system for the production of recombinant hEGF for tissue engineering applications.

Keywords: Human epidermal growth factor; Nicotiana benthamiana; Plant-produced protein; Recombinant protein; Transient expression.