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- Title
Large-scale production of biologically active human keratinocyte growth factor-2.
- Authors
Wu, Xiaoping; Tian, Haishan; Huang, Yadong; Wu, Sixian; Liu, Xiaoju; Wang, Cong; Wang, Xiaojie; Huang, Zhifeng; Xiao, Jian; Feng, Wenke; Li, Xiaokun
- 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.
- Subjects
KERATINOCYTES; GROWTH factors; ESCHERICHIA coli; RECOMBINANT proteins; AFFINITY chromatography; HEPARIN; CATIONS; GENETIC vectors; WESTERN immunoblotting
- Publication
Applied Microbiology & Biotechnology, 2009, Vol 82, Issue 3, p439
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-008-1782-x