Zhang Xiaolin, Jiang Rubin, Lin Henry, Xu Sen
Biologics Process Research & Development, Process Research & Development, Merck & Co., Inc., Kenilworth, New Jersey.
Biologics Development, Bristol-Myers Squibb Co., Pennington 08534, NJ.
Biotechnol Prog. 2020 Jul;36(4):e2975. doi: 10.1002/btpr.2975. Epub 2020 Feb 21.
Media components play an important role in modulating cell metabolism and improving product titer in mammalian cell cultures. To sustain cell productivity, highly active oxidative metabolism is desired. Here we explored the effect of tricarboxylic acid (TCA) cycle intermediates supplementation on lactate metabolism and productivity in Chinese hamster ovary fed-batch cultures. Direct addition of 5 mM alpha-ketoglutarate (α-KG), malic acid, or succinic acid in the basal medium did not have any significant impact on culture performance. On the other hand, feeding α-KG, malic acid, and succinic acid in the stationary phase, either as a single solution or as a mixture, significantly improved lactate consumption, reduced ammonium accumulation, and led to higher cell specific productivity and antibody titer (~35% increase for the best condition). Delivering those intermediates as an acidic solution for pH control eliminated CO sparging and accumulation. Feeding TCA cycle intermediates was also demonstrated to be superior to feeding lactic acid or pyruvic acid in titer improvement. Taken together, feeding TCA cycle intermediates was effective in improving lactate consumption and increasing product titer, which is likely due to enhanced oxidative metabolism in an extended duration.
培养基成分在调节哺乳动物细胞培养中的细胞代谢和提高产物滴度方面发挥着重要作用。为了维持细胞生产力，需要高度活跃的氧化代谢。在此，我们探讨了补充三羧酸（TCA）循环中间体对中国仓鼠卵巢细胞补料分批培养中乳酸代谢和生产力的影响。在基础培养基中直接添加5 mM的α-酮戊二酸（α-KG）、苹果酸或琥珀酸对培养性能没有任何显著影响。另一方面，在稳定期添加α-KG、苹果酸和琥珀酸，无论是单一溶液还是混合物，都能显著提高乳酸消耗，减少铵积累，并导致更高的细胞比生产力和抗体滴度（最佳条件下提高约35%）。以酸性溶液形式输送这些中间体以控制pH值，消除了二氧化碳的鼓泡和积累。在提高滴度方面，添加TCA循环中间体也被证明优于添加乳酸或丙酮酸。综上所述，添加TCA循环中间体可有效改善乳酸消耗并提高产物滴度，这可能是由于在延长的时间内氧化代谢增强所致。
