To increase the productivity of rCHO cells, many cell engineering approaches have been demonstrated that over-express or knockout a specific gene to achieve increased titers. In this work, we present an alternate approach, based on the concept of evolutionary adaptation, to achieve cells with higher titers.rCHO cells, producing a monoclonal antibody, are adapted to ER-stress, by continuous culturing under increasing concentration of tunicamycin. A sustained higher productivity of at-least 2-fold was achieved in all the clones, in a concentration-dependent manner. Similarly, a 1.5-2 fold increase in final titers was also achieved in the batch culture. Based on metabolic analysis of the adapted cells, a fed-batch process was designed where significantly higher titersare achieved as compared to control. Metabolic flux analysis is employed in addition with gene expression analysis of key genes to understand the basis of increased performance of the adapted cells.Overall, this work illustrates how process modifications and cellular adaptation can be used in synergy to drive up product titers.

中文翻译：

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通过优化的分批补料工艺和ER胁迫适应性结合提高rCHO克隆的效价和生产率。

为了提高rCHO细胞的生产力，许多细胞工程学方法已被证明过表达或敲除特定基因以实现更高的滴度。在这项工作中，我们提出了一种基于进化适应性概念的替代方法，以实现具有更高滴度的细胞。通过在不断增加的衣霉素浓度下连续培养，产生单克隆抗体的rCHO细胞可以适应内质网应激。在所有克隆中，都以浓度依赖的方式实现了至少2倍的持续较高生产率。同样，分批培养中最终滴度也提高了1.5-2倍。基于适应细胞的代谢分析，设计了分批补料工艺，与对照相比，滴定度明显更高。