CHO细胞无血清培养基的工艺优化

Optimization of Serum-free Medium of CHO Cells

本文以表达人肿瘤坏死因子受体-人Ig G-Fc抗体融合蛋白的CHO细胞为研究对象,通过对影响其细胞密度、活力及蛋白表达量和聚体百分比的进行了3个大类因素的考察,即水解物:0.5%酵母提取物,0.5%谷类水解物,0.5%大豆蛋白水解物,0.5%植物蛋白胨;脂类物质:10 mmol/L磷脂酰胆碱,10 mg/L乙醇胺,0.05%脂肪乳剂;激素类物质:10μmol/L氢化可的松(肾上腺皮质激素),10 mg/L胰岛素,20 n M孕酮(类固醇激素)。结果表明,水解物中0.5%酵母水解物,脂类物质中10 mg/L乙醇胺,激素类物质中10 mg/L胰岛素对提高CHO细胞密度、活力和蛋白表达量和降低聚体含量具有重要促进作用。

In this study,CHO cell,which can express antibody fusion protein of human tumor necrosis factor receptor-Fc- people Ig G,was used as research object. The effect of three important factors on the cell density,activity,expression level and percentage of polymers were investigated,including： the hydrolysate： 0. 5% yeast extract,0. 5% cereal hydrolysate,0. 5% soyprotein hydrolysate,0. 5% phytone; the lipid： 10 mmol / L phosphatidyl choline,10 mg / L ethanolamine,0. 05% intralipid; the hormone substances： 10 μmol / L hydrocortisone（ adrenocortical hormone）,10 mg / L insulin,20 n M progesterone（ steroid hormones）. The results showed that 0. 5% yeast hydrolysate,10 mg / L ethanolamine and0. 05% intralipid in lipid,10 mg / L insulin can significantly improve the density,activity and the protein expression level of CHO cells.