Nilsang Suthasinee, Kumar Ashok, Rakshit Sudip Kumar
Department of Food Engineering and Bioprocess Technology, School of Environment, Resources and Development, Asian Institute of Technology, Pathumthani, Thailand.
Appl Biochem Biotechnol. 2008 Dec;151(2-3):489-501. doi: 10.1007/s12010-008-8225-0. Epub 2008 Aug 12.
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.
多年来，人们一直在研究用于提高哺乳动物细胞培养中细胞群体生长和蛋白质生产力的工艺开发与优化。在本研究中，使用工作体积为4毫升的六孔微量滴定板系统研究了杂交瘤细胞的行为。将小鼠杂交瘤细胞系D2和2C83G2接种于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）循环的一个组成部分，由谷氨酰胺经谷氨酸生成。为研究其对细胞生长、代谢和单克隆抗体（mAb）产生的影响，在培养开始时以及另一组在72小时后，在两种培养基中均添加2 mM α-酮戊二酸（pH 7.2）。在无血清培养基中培养的D2细胞观察到高细胞密度，而2C83G2细胞系在含血清培养基中显示出高细胞密度。然而，在无血清培养基中培养的两种细胞系生长8天时活力均高于70%。补充2 mM α-酮戊二酸可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产生。与72小时后添加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产生方面产生了更好的结果。然而，72小时后添加优于根本不添加。这表明α-酮戊二酸对抗体的产生和释放具有积极作用。
