文献类型：期刊文献

中文题名：CHO细胞培养过程中柠檬酸积累特性研究

英文题名：Citrate Accumulation Characteristics in CHO Cell Culture Process

作者：何永梅[1];张欣然[1];赵亮[1];谭文松[1];叶倩[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室、上海生物制造产业省部共建协同创新中心,上海200237

年份：2025

卷号：45

期号：4

起止页码：15

中文期刊名：中国生物工程杂志

外文期刊名：China Biotechnology

收录：;北大核心:【北大核心2023】；

语种：中文

中文关键词：CHO细胞;TCA循环;柠檬酸;温度;葡萄糖浓度;抗体生产

外文关键词：CHO cells;TCA cycle;Citrate;Glucose concentration;Antibody production

摘要：目的:探究中国仓鼠卵巢细胞(Chinese Hamster Ovary cell,CHO cell)流加培养过程中柠檬酸积累现象及特征,为认识和调控柠檬酸积累奠定基础。方法:以CHO细胞为研究对象,采用实验室前期建立的流加培养工艺,考察流加培养过程中柠檬酸积累特征,探究培养工艺参数(温度和葡萄糖浓度)对柠檬酸积累的影响,建立关键代谢参数[平均葡萄糖比消耗速率(Q_(Gluc))、乳酸比生成速率(q_(Lac))、抗体比生成速率(q_(p))和三羧酸循环(tricarboxylic acid cycle,TCA cycle)的总碳流量]与柠檬酸积累的相关性。结果:温度影响CHO细胞流加培养过程中柠檬酸积累,较低温度(31℃)条件下柠檬酸比生成速率显著增大;葡萄糖浓度对柠檬酸积累过程的影响有限,葡萄糖浓度控制在2~55 mmol/L的范围内不影响柠檬酸积累过程;通过考察多批不同培养工艺参数条件下的柠檬酸积累过程,发现柠檬酸比生成速率与葡萄糖比消耗速率、抗体比生成速率、TCA循环总碳流量呈正相关,与乳酸的比生成速率呈负相关。结论:柠檬酸在细胞外积累可能反映营养物质供给充足,细胞TCA循环代谢旺盛,细胞抗体生产能力增强。
Objective:The accumulation and characteristics of citrate in CHO fed-batch culture were investigated to provide a basis for understanding and regulating the accumulation of citrate.Methods:Using CHO cells as the research object,the flow addition culture process established in the laboratory was used to investigate the accumulation characteristics of citrate during the fed-batch culture.The effects of cultivation process parameters(temperature and glucose concentration)on citrate accumulation were investigated.The correlation between key metabolic parameters(Q_(Gluc),q_(Lac),q_(p),and total carbon flux of the TCA cycle)and citrate accumulation was established.Results:Temperature affected the accumulation of citrate during the fed-batch culture of CHO cells,and the specific rate of citrate formation was significantly increased at lower temperature(31℃).The effect of glucose concentration on the accumulation process of citrate was limited,and the control of glucose concentration in the range of 2~55 mmol/L did not affect the accumulation process of citrate.The accumulation process of citrate under different cultivation process parameters was investigated in multiple batches.The specific production rate of citrate is positively correlated with the specific consumption rate of glucose,the specific production rate of antibodies,and the total carbon flux of the TCA cycle,while negatively correlated with the specific production rate of lactate.Conclusions:The above correlation indicates that extracellular citrate accumulation may reflect sufficient nutrient supply,vigorous intracellular TCA cycle metabolism,and enhanced antibody production capacity.