Effect of α-Ketoglutarate on Monoclonal Antibody Production of Hybridoma Cell Lines in Serum-Free and Serum-Containing Medium
Effect of α-Ketoglutarate on Monoclonal Antibody Production of Hybridoma Cell Lines in Serum-Free and Serum-Containing Medium
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DOI：
10.1007/s12010-008-8225-0
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发表时间：
2008-12-01
影响因子：
3
通讯作者：
Rakshit, Sudip Kumar
中科院分区：
文献类型：
--
作者：
Nilsang, Suthasinee;Kumar, Ashok;Rakshit, Sudip Kumar
用于提高哺乳动物细胞培养中的群体生长和蛋白质生产率的工艺开发和优化已经研究了多年。在这项研究中，杂交瘤细胞的行为进行了研究，使用六孔微量滴定板系统的工作体积为4毫升。将小鼠杂交瘤细胞系D2和2C 83 G2接种在无血清和含血清培养基中并培养8天。α-酮戊二酸是三羧酸（TCA）循环的组成部分，由谷氨酰胺通过谷氨酸产生。为了研究其对细胞生长、代谢和单克隆抗体（mAb）产生的影响，在培养开始时向两种培养基中加入2 mM α-酮戊二酸（pH 7.2），并在72 h后在另一组培养基中加入。D2细胞在无血清培养基中培养时细胞密度较高，而2C 83 G2细胞在含血清培养基中培养时细胞密度较高。然而，在无血清培养基中培养的两种细胞系在生长8天时的活力均高于70%。2 mM α-酮戊二酸的补充支持两种杂交瘤细胞系在无血清和含血清培养基中的细胞生长和mAb生产。与72小时后补充α-酮戊二酸相比，在分批培养开始时添加α-酮戊二酸在细胞生长和mAb生产方面产生更好的结果。然而，在72小时后添加比根本不添加好。这表明α-酮戊二酸对抗体的产生和释放具有积极作用。
Process development and optimization for increase population growth and protein productivity in mammalian cell culture have been studied for many years. In this study, the behavior of hybridoma cells was investigated using six-well micro-titer plate systems with a working volume of 4 ml. Mouse hybridoma cell lines D2 and 2C83G2 were seeded in serum-free and serum-containing media and cultured for 8 days. alpha-Ketoglutarate is an integral component of the tricarboxylic acid (TCA) cycle and is produced from glutamine via glutamate. To study its effect on cell growth, metabolism, and monoclonal antibody (mAb) production, 2 mM alpha-ketoglutarate (pH 7.2) was added in both media at the beginning of the cultivation and in another set after 72 h. High cell density was observed in D2 cell culturing in serum-free medium, while 2C83G2 cell line showed high cell density in serum-containing medium. However, both cell lines cultured in serum-free medium gave viability above 70% when grown for 8 days. The supplement of 2 mM alpha-ketoglutarate supported cell growth and mAb production of both hybridoma cell lines in serum-free and serum-containing medium. The addition of alpha-ketoglutarate at the beginning of the batch cultivation gave better result in cell growth and mAb production as compared to alpha-ketoglutarate supplementation after 72 h. However, addition after 72 h was better than no addition at all. This indicates that alpha-ketoglutarate have a positive effect on production and release of antibody.