Purification of recombinant human fibroblast growth factor-2 from one-liter fermetation broth

Abstract

Fibroblast Growth Factor - 2 (FGF-2), also known as basic FGF is a multi-functional protein that regulates the proliferation and differentiation of multiple types of cell. Recombinant human FGF-2 (rhFGF-2) is currently used in medicine, cosmetics, stem-cell culture, etc. In this study, we conducted one-liter scale fermentation using Escherichia coli strain that carries recombinant vector harboring FGF-2 coding gene to produce a large amount of FGF-2 protein. The evaluation of the fermentation efficacy was based on the growth curve, dry cell weight and amount of FGF-2 obtaining from one-liter fermentation. After fermentation, cell mass was lyzed by high pressure. Then, the FGF-2 in supernatant was purified by cation exchange and heparin-affinity chromatography. The purity and efficiency of the purification process were estimated by Bradford, silver staining and densitometry using Quantity-One software. The result showed that in one-liter fermentation, we obtained 5.2 g/liter dry cell-mass and 230 mg/liter FGF-2. The purity of FGF-2 was about 97.1 % and the purification efficiency was  above 46.08 %.