Large-scale production of biologically active human keratinocyte growth factor-2.
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.
Wu
Xiaoping
X
Institute of Tissue Transplantation and Immunology, Jinan University, Guangzhou, Guangdong, 510632, China.
Tian
Haishan
H
Huang
Yadong
Y
Wu
Sixian
S
Liu
Xiaoju
X
Wang
Cong
C
Wang
Xiaojie
X
Huang
Zhifeng
Z
Xiao
Jian
J
Feng
Wenke
W
Li
Xiaokun
X
D018533:Biomass; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005285:Fermentation; D051526:Fibroblast Growth Factor 10 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008934:Mitogens / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D051526:Fibroblast Growth Factor 10; D008934:Mitogens; D011994:Recombinant Proteins