运用DOE法优化表达抗人类表皮生长因子受体2单克隆抗体细胞用培养基

Optimization of culture medium for production of monoclonal antibody against human epidermal growth factor receptor 2 by using design of experiment

目的运用实验设计(design of experiments,DOE)方法优化表达抗人类表皮生长因子受体2(human epidermal growth factor receptor 2,HER2)单克隆抗体细胞用培养基,以提高抗体表达量。方法利用Minitab软件进行DOE,考察不同基础培养基、补料培养基对工程细胞株(CHO-DG44)的细胞密度和抗HER2单克隆抗体表达的影响。结果经过DOE方法优化,在最优培养基组合中工程细胞株的最高活细胞密度(peak viable cell density,PVCD)达296×105个/ml,抗体表达量最高达2.07 g/L,比DOE优化之前最高试验组表达量(1.20 g/L)提高了72%,所表达抗体的质量与原研对照品非常一致。结论得到1组较优的基础培养基与补料培养基配比与组合,该培养基组合提高了细胞密度与抗体表达量,且其适合抗HER2单克隆抗体生产使用,表明DOE是一种短期快速提高抗体表达行之有效的方法。

Objective To optimize the culture medium for production of monoclonal antibody（Mc Ab） against human epidermal growth factor receptor 2（HER2） by using design of experiments（DOE） so as to increase the expression level of Mc Ab. Methods The effects of various basal media and feeding media on cell density and expression of Mc Ab against HER2 in engineering cell were investigated by DOE with Minitab software. Results After optimization by DOE, the viable cell density of recombinant cell strain reached 296 × 105 cells / ml at most. However, the expression level of antibody titer reached 2. 07 g / L at most, which increased by 72% compared with that before optimization（1. 20 g / L）.The quality of antibody was similar to that of antibody developed previously as control. Conclusion An optimized combination of basal media and feeding media was obtained, by which the cell density and antibody expression level increased. The optimized combination was suitable for production of Mc Ab against HER2, indicating DOE as an effective method for rapidly increasing the expression level of antibody within a short time period.