大肠杆菌中表达的rhHGF-α以包涵体形式存在，表达量约占菌体总蛋白的25％。为了建立一条简便的分离纯化生产工艺，我们研究了包涵体溶解及复性条件，并进行了活性测定。结果表明，用含有4 mol/L尿素的缓冲液洗涤包涵体，8 mol/L尿素溶解包涵体后的上清，经过SephacrylS-200凝胶过滤、复性后，得到纯度达90％的rhHGF-a，SDS-PAGE测定相对分子质量为52 000。与rhHGF-β体外共复性后，完整的rhHGF具有明显刺激原代培养大鼠肝细胞生长的活性。%Recombinant human hepatocyte growth factor a chain produces as inclusion body in E. coli DH5a with 25% expression level. The solubilization and renaturation conditions of rhHGF-a were studied to develop a way to facilitate the purification of inclusion body. Highly purified inclusion body was obtained via several steps including washing with 4 mol/L urea buffers, solubilization in 8 mol/L urea and Sephacryl S-200 chromatography. Relative molecular weight of the rhHGF-a determined by SDS-PAGE was 52 000. After renaturation with rhHGF-β, the whole HGF shows biological activity of increasing the growth of rat hepatocyte in primary culture.
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