Fed-batch培养中补料及接种密度对RABV-G蛋白表达量的影响

Effect of feeding and inoculation density on the expression of Glycoprotein in Fed-batch culture

目的研究流加(Fed-batch)培养中补料及接种密度对狂犬病毒糖蛋白G(RABV-G)蛋白表达量的影响,初步建立RABV-G蛋白在重组中国仓鼠卵巢(CHO)细胞中Fed-batch培养的较优工艺方案。方法复苏培养1支能够稳定表达狂犬病毒RABV-G蛋白的重组CHO细胞,传代扩增后进行6种方案Fed-batch培养,从Fed-batch培养的第3天开始监测培养上清中目的蛋白表达量和细胞生长参数,第14天结束培养,分析比较不同策略所产生的目的蛋白表达量。结果本研究方案6中5%的Feed1补料及0.5×10~6cells/ml的起始接种密度在Fed-batch培养第14天,糖蛋白表达量达150 mg/L,显著高于其余5种方案,确立为优选方案。结论本研究建立了RABV-G蛋白在重组CHO细胞中流加(Fed-batch)培养的较优工艺方案,为重组狂犬病毒疫苗生产工艺研究提供基础。

Objective To establish the optimal technology of Fed-batch culture of rabies virus glycoprotein G(RABV-G) in recombinant Chinese hamster ovary(CHO) cells. Methods Recombinant CHO cells stably expressing rabies virus G glycoprotein were cultured by resuscitated culture, and Fed-batch culture was carried out after passage and expansion, the expression of target protein in the supernatant and cell growth parameters were monitored from the 3rdday of Fed-batch Culture, and at the 14thday of Culture, the expression of target protein produced by different strategies were analyzed and compared. Results In this study, 5% feed 1 and 0.5×10~6cells/ml initial inoculum density in the sixth protocol were significantly higher than those in the other five protocols at 150 mg/L on the 14thday of Fed-batch culture. Conclusion The optimal technology of Fed-batch culture of rabies virus glycoprotein in recombinant CHO cells was established in this study,which provided a basis for the study of the production technology of recombinant rabies virus vaccine.