Xiao Shang, Li Qiang, Jiang Jinlong, Huo Chengxiao, Chen Hao, Guo Meijin
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Shanghai Biological Products Research Institute Co., Ltd., Shanghai 200051, China.
Bioengineering (Basel). 2023 Mar 14;10(3):357. doi: 10.3390/bioengineering10030357.
Currently, the assessment of process robustness is often time-consuming, labor-intensive, and material-intensive using process characterization studies. Therefore, a simple and time-saving method is highly needed for the biopharmaceutical industry. Apoptosis is responsible for 80% of Chinese hamster ovary (CHO) cell deaths and affects the robustness of the cell culture process. This study's results showed that a more robust process can support cells to tolerate apoptosis for a longer time, suggesting that the robustness of the process could be judged by the ability of cells to resist apoptosis. Therefore, it is necessary to establish a rapid method to detect the apoptosis of CHO cells. In trying to establish a new method for detecting apoptosis in large-scale cell cultures, glucose withdrawal was studied, and the results showed that CHO cells began to apoptose after glucose was consumed. Then, the concentration of extracellular potassium increased, and a prolongation of apoptosis time was observed. Further study results showed that the process with poor robustness was associated with a higher proportion of apoptosis and extracellular potassium concentration, so potassium could be used as a biochemical index of apoptosis. The strategy we present may be used to expedite the assessment of process robustness to obtain a robust cell culture process for other biologics.
目前，使用工艺特性研究来评估工艺稳健性通常既耗时、费力又耗材。因此，生物制药行业迫切需要一种简单且省时的方法。细胞凋亡导致80%的中国仓鼠卵巢（CHO）细胞死亡，并影响细胞培养过程的稳健性。本研究结果表明，更稳健的工艺能够支持细胞更长时间地耐受细胞凋亡，这表明工艺的稳健性可以通过细胞抵抗细胞凋亡的能力来判断。因此，有必要建立一种快速检测CHO细胞凋亡的方法。在尝试建立一种在大规模细胞培养中检测细胞凋亡的新方法时，研究了葡萄糖剥夺情况，结果表明CHO细胞在葡萄糖耗尽后开始凋亡。然后，细胞外钾浓度升高，且观察到细胞凋亡时间延长。进一步的研究结果表明，稳健性较差的工艺与较高比例的细胞凋亡和细胞外钾浓度相关，因此钾可作为细胞凋亡的生化指标。我们提出的策略可用于加快工艺稳健性评估，从而为其他生物制品获得稳健的细胞培养工艺。
