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Production of Functional Hepatocyte Growth Factor (HGF) in Insect Cells Infected with an HGF‐Recombinant Baculovirus in a Serum‐Free Medium
Author(s) -
Wang MinYing,
Yang YaHuey,
Lee HsuanShu,
Lai SuYuan
Publication year - 2000
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp990143t
Subject(s) - hepatocyte growth factor , sf9 , proteases , cell culture , protease , recombinant dna , biology , microbiology and biotechnology , chemistry , biochemistry , spodoptera , enzyme , receptor , genetics , gene
Three insect cell lines, SL‐7B cells derived from Spodoptera litura, Sf9, and High Five (Hi‐5) cells, were used for the production of pro‐hepatocyte growth factor (pro‐HGF). Cells were cultured and then infected with a recombinant HGF‐containing baculovirus in a serum‐free medium. In SL‐7B cells, pro‐HGF is synthesized and excreted from the cells and late in infection is converted to a heterodimeric form of HGF even when the cells are grown in serum free medium. Conversion of a single‐chain form of HGF (pro‐HGF) into an HGF heterodimer was unexpected, as pro‐HGF is normally cleaved by a serum protease called HGF activator. The proliferation activity of heparin‐affinity‐purified HGF from serum‐free culture supernatant of SL‐7B cells is comparable to that obtained from HGF converted by serum proteases, suggesting that SL‐7B cells produce a functionally analogous protease to correctly process pro‐HGF. This work reports, for the first time, on the feasibility of properly processing pro‐HGF to form functional HGF by proteases from invertebrate cells in serum‐free media. Avoiding the supplementation of sera provides the advantages of a low production cost, zero contamination of infectious agents from sera, and simple downstream product purification. Experimental results further demonstrate that the conversion of pro‐HGF by insect cells is cell‐line‐dependent, because proteases in Hi‐5 or Sf9 cells could not process pro‐HGF as efficiently and properly as those in SL‐7B cells.