CHO细胞表达抗血管内皮生长因子单克隆抗体工艺优化

抗血管内皮生长因子单克隆抗体(VEGF-MA)能够抑制肿瘤生长,具有良好的市场前景。本研究利用一株重组中国仓鼠卵巢细胞(Chinese hamster ovary cell,CHO)细胞株表达VEGF-MA。首先对培养基种类进行优化,筛选最优的基础培养基、补料培养基和外源添加物。研究结果表明:最有利于重组CHO细胞株表达VEGF-MA的基础培养基、补料培养基和外源添加物分别为ActiCHO P Powder CD、CD Efficient Feed C AG和Sheff-CHO Plus PF ACF。利用这一培养基配比在摇瓶和3 L发酵罐中培养重组CHO细胞,VEGF-MA产量均可达到3.20 g/L。在3 L发酵罐中进一步优化培养条件,结果表明:最优的接种密度、pH、溶解氧浓度、前期培养温度和后期培养温度分别为1.0×10^6个/mL、7.10、40%、36.5℃和34℃,此时的VEGF-MA产量能够达到4.10 g/L。VEGF-MA质量指标均处于标准范围内:电荷异质性、糖基化水平和蛋白纯度分别为26.1%、59.1%和95.1%。

Culture condition optimization for expression of anti-human VEGF monoclonal antibody by CHO cell

Anti-human VEGF monoclonal antibody(VEGF-MA)can inhibit tumor growth.Therefore,it has good market prospects.In this study,a recombinant Chinese hamster ovary cell(CHO)was used to express VEGF-MA.Medium were firstly optimized,aiming at selecting optimal basal media,feeding media and external additive.It was concluded that the best basal media,feeding media and external additive for the expression of VEGF-MA by CHO were ActiCHO P Powder CD,CD Efficient Feed C AG and Sheff-CHO Plus PF ACF,respectively.By using the optimal medium,the yield of VEGF-MA in 3 L fermentor could reach 3.20 g/L.In addition,the culture conditions in 3 L fermentor were optimized.The optimal initial density,pH,dissolved oxygen,initial temperature and shifted temperature were 1.0×10^6/mL,7.10,40%,36.5℃and 34℃,respectively.Under the optimized conditions,the yield of VEGF-MA in 3 L fermentor could reach 4.10 g/L.The VEGF-MA quality indices were located in standard range:charge heterogeneity,glycosylation level and protein purity were 26.1%,59.1%and 95.1%,respectively.