二氧化碳分压和渗透压升高对CHO细胞维持期生长、代谢和产物表达的影响  

Effects of Raising pCO_2 and Osmolality on the Growth,Metabolism and Productivity of CHO Cells at Maintenance Phase

作　　者：王晨[1] 汪嘉琪 赵亮[1,2] 范里 刘旭平[1,2] 陈敏 张理想[2] 谭文松 WANG Chen;WANG Jia-qi;ZHAO Liang;FAN Li;LIU Xu-ping;CHEN Min;ZHANG Li-xiang;TAN Wen-song(State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237;Bio-engine Technology Co., Ltd., Shanghai 200237)

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237 [2]上海倍谙基生物科技有限公司,上海200237

出　　处：《生物技术通报》2018年第3期217-224,共8页Biotechnology Bulletin

基　　金：国家自然科学基金项目(21106045);国家重大专项(2013ZX10004003-003-003)

摘　　要：为了深入了解培养体系中二氧化碳分压(pCO_2)和渗透压对中国仓鼠卵巢(CHO)细胞产物表达的影响。考察了不同pCO_2和渗透压条件下CHO细胞的密度维持、代谢、产物表达量和产品质量(糖型修饰和电荷异质化)的情况。pCO_2和渗透压升高均对最终抗体表达产生不利的影响,其中渗透压升高对细胞后期活性维持不利,而pCO_2升高则对产物表达期抗体比生成速率产生负面影响。通过对过程中胞内外代谢物的研究,发现pCO_2和渗透压升高均会降低碳源有效利用率。在产物的质量方面,当渗透压升高时,产物的糖基化水平下降,产物电荷分析中的中性电荷含量比例升高;而当pCO_2升高时,产物的糖型分布无明显变化,但其中中性电荷抗体含量降低。产物表达期的pCO_2和渗透压的升高对细胞维持、物质能量代谢及产物表达存在诸多不利影响,因此应尽量避免培养体系中的CO_2累积和渗透压升高的情况发生,以确保培养规模放大的正常进行。The aim of this study is to understand the effects of pCO_2 and osmolality on the monoclonal antibody（m Ab）productionduring the maintenance phase of Chinese hamster ovary（CHO）cell cultures. Cell maintenance,metabolism,antibody quantity and qualityat different pCO_2 and osmolality levels were investigated during cell maintenance. It was found that the raise of pCO_2 and osmolality broughtnegative impacts on final antibody production. The increase of osmolality damaged the cell maintenance,while the increase of pCO_2 downregulated the productivity of CHO cells. Through comparing the extracellular and intracellular metabolites,the increase of pCO_2 and osmolalityresulted in the decrease of carbon source utilization. Besides,on the antibody quality,the increase of osmolality decreased the glycosylationbut raised the ratio of neutral charge. Moreover,the increase of pCO_2 mainly enhanced the charge heterogeneity but there were no changeson the sugar type distribution of the products. Conclusively,the increase of pCO_2 and osmolality during the phase of antibody expressionhad unfavorable effects on the cell maintenance,both material and energy metabolism,and the expression of product antibody;thus theaccumulation of CO_2 and osmolality elevation should be avoided as much as possible in order to ensure the normal proceeding of large-scaleculture.

关 键 词：二氧化碳分压 渗透压 CHO细胞 单克隆抗体 代谢 

分 类 号：Q25[生物学—细胞生物学]