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低CO2部分压力将CHO细胞引导到一个有缺陷的代谢状态

Liang Zhao¹, Chen Wang¹, Jiaqi Wang²

¹State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, P. O. Box 309#, Shanghai, 200237, People's Republic of China.

Biotechnology letters

|June 15, 2023

概括

低二氧化碳部分压力 (pCO2) 损害了中国汉姆斯特卵巢 (CHO) 细胞代谢和单克隆抗体的产生. 添加酸盐可以部分恢复有氧代谢,为设计质量 (QbD) 的过程控制提供信息.

科学领域:

生物技术是生物技术.
细胞培养细胞培养
生物工艺工程生物工艺工程

背景情况:

在大型动物细胞培养中,二氧化碳的积累是有害的.
反应堆运行不当可能导致有害的低二氧化碳部分压力 (pCO2) 条件.
了解pCO2的影响对于优化生物工艺和确保产品质量至关重要.

研究的目的:

为了研究超低pCO2对中国汉姆斯特卵巢 (CHO) 细胞代谢和生产力的影响.
根据设计质量 (QbD) 指南,提供用于确定设计空间的二氧化碳控制的数据.
确定减轻低pCO2的不良影响的策略.

主要方法:

通过头部空气净化诱导超低pCO2 (ULC) 条件.
分析单克隆抗体的产生和有氧代谢活动.
使用细胞内代谢学来评估代谢状态.
测量细胞内pH值和乳酸脱酶活性.
采用半经验数学模型进行pCO2预测和控制.

主要成果:

ULC条件降低了单克隆抗体的产生和有氧代谢活动.
细胞内代谢学显示,在ULC下,有氧葡萄糖代谢效率较低.
细胞内pH值升高和乳酸脱酶活性增加表明pyruvate缺乏.
此外,酸盐部分缓解了缺陷的有氧代谢.
开发了一个数学模型来预测和控制极端的pCO2.

结论:

低pCO2诱导CHO细胞中一个有缺陷的代谢状态.
与pCO2,乳酸盐和pH相关的预测模型有助于强大的代谢控制.
这些发现支持确定在CHO细胞培养中控制CO2的QbD设计空间.

关键词:

在CHO细胞中.部分压力 CO2 的部分压力.细胞内部的pH值乳酸脱酶的使用方法乳酸盐代谢乳酸代谢代谢学代谢学代谢学

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