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Large-scale production of biologically active human keratinocyte growth factor-2

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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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The work was supported by grants from the National High-Tech Research and Development Program of China (2007AA02Z110) and the Science Foundation of Zhejiang Province of China (No. Z205755).

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Correspondence to Xiaokun Li.

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Xiaoping Wu and Haishan Tian contributed equally to this work.

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Wu, X., Tian, H., Huang, Y. et al. Large-scale production of biologically active human keratinocyte growth factor-2. Appl Microbiol Biotechnol 82, 439–444 (2009). https://doi.org/10.1007/s00253-008-1782-x

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  • Human keratinocyte growth factor-2
  • Large-scale production
  • Expression
  • Purification
  • Mitogenic activity