无血清培养基的优化对rhEPO-Fc融合蛋白体外活性的影响

Effect of the optimization of serum-free medium on the in vitro activity of rhEPO-Fc fusion protein

目的 :研究基础培养基I/F/D中添加不同浓度的培养基组分对rh EPO-Fc融合蛋白体外活性的影响。方法 :通过单因素实验结合Box-Behnken效应面法,筛选适合rh EPO-Fc表达的较优培养基组分。结果 :在基础培养基中添加1%F68、10%维生素、7.7 g/L腐胺、10.1 g/L乙醇胺和5.38 g/L胰岛素时,rh EPO-Fc融合蛋白的体外活性可达10 268IU/ml,比初始的基础培养基I/F/D提高了108.4%。结论 :优化后的培养基能提高rh EPO-Fc融合蛋白体外活性,降低生产成本,可为后期的中试生产提供重要参考。

Objective： To study the effects of the adding different concentrations of various components in the basal culture medium I/F/D on the in vitro activity of rhEPO-Fc. Methods： The optimal components suitable for the expression of rhEPO-Fc were screened by the single factor experimental results combined with Box-Behnken response surface method. Results： When 1% F68, 10% vitamin, 7.7 g/L putrescine, 10.1 g/L ethanolamine and 5.38 g/L insulin were added in the basal culture medium, the in vitro activity of rhEPO-Fc was increased by 108.4% up to 10 268 IU/ml compared with the results in the initial basal serum free medium. Conclusion： The in vitro activity of rhEPO-Fc can be increased when cells were incubated in the optimized culture medium, the costs for the production of rhEPO-Fc can be obviously reduced. The results of this study can provide a significant reference for pilot production of rhEPO-Fc.