Development of a Cost Effective Purification Method for Human Epidermal Growth Factor (hEGF) in Recombinant Escherichia coli

Human Epidermal Growth Factor (hEGF) with 53 amino acids and a molecular weight of 6.2 KD is a protein that stimulates the growth of various cell types. The purpose of this study was to apply a novel method for purification of the recombinant hEGF that was expressed in recombinant Escherichia coli BL21(DE3). The recombinant strain produced hEGF in form of intracellular so it was necessary to disrupt cell wall of recombinant E. coli and then purify hEGF from inclusion body. Renaturation and purification protocol, which was used in this study, was a simple and efficient one that had three stages including unfolding, purification by means of ultrafiltration and centrifugation and finally refolding of unfolded rhEGF molecules. SDS-PAGE and Reverse Phase High Performance Liquid Chromatography (RP-HPLC) were used as analytical methods to investigate the results. The purified protein was analyzed by SDS-PAGE and RP-HPLC and the results were shown more than 99% purity.