文献类型：期刊文献

中文题名：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

英文题名：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[1];赵亮[1,2];范里[1,2];刘旭平[1,2];陈敏[2];张理想[2];谭文松[1,2]

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200237;[2]上海倍谙基生物科技有限公司,上海200237

年份：2018

卷号：34

期号：3

起止页码：217

中文期刊名：生物技术通报

外文期刊名：Biotechnology Bulletin

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD_E2017_2018】；

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

语种：中文

中文关键词：二氧化碳分压;渗透压;CHO细胞;单克隆抗体;代谢

外文关键词：pCO2 ; osmolality ; CHO ceils ; monoclonal antibody ; metabolism

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.