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  • Title: Large-scale production of biologically active human keratinocyte growth factor-2.
    Author: Wu X, Tian H, Huang Y, Wu S, Liu X, Wang C, Wang X, Huang Z, Xiao J, Feng W, Li X.
    Journal: Appl Microbiol Biotechnol; 2009 Mar; 82(3):439-44. PubMed ID: 19015842.
    Abstract:
    A rapid and efficient expression and purification system has been developed for large-scale production of biologically active recombinant human keratinocyte growth factor-2 (rhKGF-2). The gene encoding human KGF-2 was cloned into the expression vector pET3c and transformed into Escherichia coli BL21(DE3)/pLys S. Under optimal conditions in a 30-l fermentor, the average bacterial yield and the average expression level of rhKGF-2 of three batches were up to 732 g and 32%, respectively. The recombinant protein was purified by cation exchange and heparin-affinity chromatography. One hundred and sixty five milligrams of pure rhKGF-2 was achieved per liter culture. A preliminary biochemical characterization of purified rhKGF-2 was performed by Western blotting and mitogenic activity analysis, and the results demonstrated that purified rhKGF-2 could react with anti-human KGF-2 antibody and stimulate the proliferation of HaCat cells.
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