Xu Ping, Dai Xiao-Ping, Graf Erica, Martel Richard, Russell Reb
Global Manufacturing and Supply, Bristol-Myers Squibb Company, 519 Route 173 West, Bloomsbury, NJ, 08804.
Biotechnol Prog. 2014 Nov-Dec;30(6):1457-68. doi: 10.1002/btpr.1957. Epub 2014 Aug 8.
A unique and nontraditional approach using glutamine and asparagine supplements for CHO-glutamine synthetase (GS) cell lines was studied. In our experiments, we found that a decrease in pH and an increase in cell death occurred in production phase of a GS cell line, leading to reduced antibody expression and lower antibody yields. The experimental results and the statistical analysis (ANOVA) indicated that additions of glutamine and asparagine in the basal and feed media were effective to buffer the cell culture pH, reduce lactate generation, maintain a higher cell viability profile, and improve antibody productivity. In bench-top bioreactors, glutamine and asparagine supplementation helped to prevent cell death, improve antibody yield, and reduce base usage. Glutamine is normally excluded from culture media for GS cell lines to prevent the bypass of selection pressure. In this study, however, the addition of glutamine did not affect cell population homogeneity, protein quality, or decrease antibody yield of two GS cell lines.
研究了一种针对CHO-谷氨酰胺合成酶(GS)细胞系使用谷氨酰胺和天冬酰胺补充剂的独特且非传统的方法。在我们的实验中,我们发现在GS细胞系的生产阶段pH值下降且细胞死亡增加,导致抗体表达减少和抗体产量降低。实验结果和统计分析(方差分析)表明,在基础培养基和补料培养基中添加谷氨酰胺和天冬酰胺可有效缓冲细胞培养的pH值,减少乳酸生成,维持较高的细胞活力水平,并提高抗体生产力。在台式生物反应器中,补充谷氨酰胺和天冬酰胺有助于防止细胞死亡,提高抗体产量,并减少碱的用量。通常在GS细胞系的培养基中不添加谷氨酰胺以防止选择压力的绕过。然而,在本研究中,添加谷氨酰胺并未影响两种GS细胞系的细胞群体同质性、蛋白质质量或降低抗体产量。