无血清培养基的优化对rhEPO-Fc融合蛋白体外活性的影响
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摘 要 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rhEPO-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rhEPO-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，rhEPO-Fc融合蛋白的体外活性可达10 268 IU/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rhEPO-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。
关键词 无血清培养基 腐胺 乙醇胺 胰岛素
中图分类号：Q813.11 文献标识码：A 文章编号：1006-1533（2015）23-0074-06
Effect of the optimization of serum-free medium on the in vitro activity of rhEPO-Fc fusion protein
WANG Yan*， JIANG Ying， LI Ying， ZHOU Yongchun
（Shanghai Chemo Wanbang Biopharma， Shanghai 201506， China）
ABSTRACT 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....
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