Kyriakopoulos Sarantos, Kontoravdi Cleo
Centre for Process Systems Engineering, Department of Chemical Engineering, Imperial College London, London, SW7 2AZ, United Kingdom.
Biotechnol Bioeng. 2014 Dec;111(12):2466-76. doi: 10.1002/bit.25319. Epub 2014 Oct 21.
A methodology to calculate the required amount of amino acids (a.a.) and glucose in feeds for animal cell culture from monitoring their levels in batch experiments is presented herein. Experiments with the designed feeds on an antibody-producing Chinese hamster ovary cell line resulted in a 3-fold increase in titer compared to batch culture. Adding 40% more nutrients to the same feed further increases the yield to 3.5 higher than in batch culture. Our results show that above a certain threshold there is no linear correlation between nutrient addition and the integral of viable cell concentration. In addition, although high ammonia levels hinder cell growth, they do not appear to affect specific antibody productivity, while we hypothesize that high extracellular lactate concentration is the cause for the metabolic shift towards lactate consumption for the cell line used. Overall, the performance of the designed feeds is comparable to that of a commercial feed that was tested in parallel. Expanding this approach to more nutrients, as well as changing the ratio of certain amino acids as informed by flux balance analysis, could achieve even higher yields.
本文提出了一种通过监测分批实验中氨基酸（a.a.）和葡萄糖的水平来计算动物细胞培养饲料中所需氨基酸和葡萄糖量的方法。在产生抗体的中国仓鼠卵巢细胞系上使用设计好的饲料进行实验，结果表明与分批培养相比，滴度提高了3倍。在相同饲料中额外添加40%的营养物质，产量进一步提高，比分批培养高出3.5倍。我们的结果表明，超过一定阈值后，营养物质添加量与活细胞浓度积分之间不存在线性相关性。此外，虽然高氨水平会阻碍细胞生长，但它们似乎不影响特定抗体的生产率，而我们推测高细胞外乳酸浓度是所用细胞系代谢转向乳酸消耗的原因。总体而言，设计饲料的性能与同时测试的商业饲料相当。将这种方法扩展到更多营养物质，并根据通量平衡分析改变某些氨基酸的比例，可能会实现更高的产量。
