单克隆抗体Mab-A生产细胞Sp2/0缩小模型培养优化研究

Sp2/0是一种生产单克隆抗体的常用细胞株。本研究首先在批次培养模式中对适合Sp2/0细胞生长的5种基础培养基、摇床转速、培养温度、二氧化碳浓度、微量元素和GlutaMAX ^TM替换谷氨酰胺等影响因素进行了筛选研究。结果显示Sp2/0细胞在批次培养中细胞密度最高值达到13.12×10 ⁶ cells/ml,培养时间为7天。除培养温度会导致不同的细胞生长密度和活率、进而影响培养时间外,其它因素不能导致明显的细胞生长差异。随后在流加培养模式下就14种补料组合进行了筛选,Sp2/0在流加培养模式下细胞的峰值密度可达20~30×10 ⁶ cells/ml,培养时间9天,单克隆抗体Mab-A日产量最高达到27.20mg/L。最后应用批次-反复流加培养模式培养Sp2/0细胞,该条件下峰值细胞数为50.42×10 ⁶ cells/ml,培养时间14天,每天单抗产量(141.10mg/L)是流加培养的5.19倍。这些研究结果为Sp2/0细胞规模化生产单克隆抗体奠定了一定基础。

Study on the Optimal Scale-down Model for Cell Growth and Mab-A Production of Sp2/0 Cells

Five different kinds of basal media were screened in batch mode to find out the optimal basal medium for cell growth of Sp2/0. The peak viable cell density in batch mode was 13.12×10 ⁶cells/ml, and the culture duration was 7 days. Various cell culture conditions including different shaking speeds, concentrations of carbon dioxide, glutamine replaced by GlutaMAX ^TM, addition of trace element and different culture temperatures were studied in batch mode as well. Few different cell growth were found with these conditions except for culture temperatures, which leaded to different peak viable cell concentrations, different viabilites and then different culture durations. Fourteen kinds of combinations of feed media were screened in fed-batch mode. The peak viable cell densities were up to 30×10 ⁶cells/ml, the culture duration were around 9 days, and the highest daily Mab-A production was 27.20mg/L. Batch re-feed mode was used in the third part of the study. The peak viable cell density was 50.42×10 ⁶cells/ml, the culture duration was 14 days, and the highest daily Mab-A production (141.10mg/L) was 5.19 fold greater than in fed-batch mode. These studies suggest batch re-feed mode is the optimal scale-down mode for cell growth and Mab-A production of Sp2/0 cells.