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以包涵体的形式在大肠杆菌中表达人血管内皮细胞生长因子(VEGF121) , 采用发酵罐进行工程菌的高密度发酵,得到菌干重 46 g/L, VEGF121 包涵体 4.5 g/L。包涵体经洗涤、溶解、超滤法复性, 得率为 81%。复性后的目标蛋白经离子交换色谱及 Sepharcry S-100 凝胶过滤色谱纯化, 目标蛋白的纯度达到 95%, 目标蛋白的总得率 31%。采用该工艺制备的 VEGF121 能刺激人脐静脉血管内皮细胞增殖, 具有天然 VEGF 生物学活性。
Human vascular endothelial cell growth factor (VEGF121) was expressed in E. coli as an inclusion body, and the high density fermentation of engineered bacteria was carried out in a fermentor to obtain a bacterial dry weight of 46 g / L and a VEGF121 inclusion body of 4.5 g / L. Inclusion body was washed, dissolved, ultrafiltration refolding, the yield was 81%. The renatured target protein was purified by ion-exchange chromatography and Sephacryl S-100 gel filtration chromatography. The purity of the target protein was 95%, and the total yield of the target protein was 31%. The VEGF121 prepared by the process can stimulate the proliferation of human umbilical vein endothelial cells and has the biological activity of natural VEGF.