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Stable expression of rhVEGF165 in Chinese hamster ovary cells
rhVEGF165在中国仓鼠卵巢细胞中的稳定表达、纯化及其生物学活性

Keywords: rhVEGF165,Chinese hamster ovary (CHO) cell,stable expression,bioreactor process,rhVEGF165 purification
血管内皮生长因子
,(VEGF165),中国仓鼠卵巢细胞,重组蛋白稳定表达,生物反应器,rhVEGF165蛋白纯化

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Abstract:

We established a stable Chinese hamster ovary (CHO-S) cell line for recombinant human VEGF165-expressing. We co-transfected GS-expression vector and rhVEGF165 expression plasmid into CHO-S cells, and selected the highest VEGF165- expressing clone as the working cell line to express VEGF165 protein. After 7-day fed-batch culture in a 5 L bioreactor and 3 steps chromatographic purification, we got the rhVEGF165 protein for series of binding and biological activity examination. The production was over 50 mg/L. The purified rhVEGF165 protein was functionally active with a half-maximal Human Umbilical Vein Endothelial Cells (HUVEC) growth-enhancing effect concentration of 1.94 ng/mL. It was slightly better than commercially available Escherichia coli expressing rhVEGF165. So we expressed successfully rhVEGF165 protein in high-level and obtained the fully active rhVEGF165 protein in large quantity.

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