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

Abstract

Fibroblast Growth Factor - 2 (FGF-2), also known as basic FGF is a multifunctional 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 QuantityOne software. The result showed that in oneliter 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 %.