Cell Culture and Fermentation Sciences, BioPharmaceuticals Development, R&D, AstraZeneca, Cambridge, UK.
Kymab Ltd, Cell Line Development, Biopharmaceutical Development, Kymab, Babraham Research Campus, Cambridge, UK.
Biotechnol Bioeng. 2021 Jun;118(6):2326-2337. doi: 10.1002/bit.27744. Epub 2021 Mar 25.
The manufacture of bispecific antibodies by Chinese hamster ovary (CHO) cells is often hindered by lower product yields compared to monoclonal antibodies. Recently, reactive oxygen species have been shown to negatively impact antibody production. By contrast, strategies to boost cellular antioxidant capacity appear to be beneficial for recombinant protein expression. With this in mind, we generated a novel hydrogen peroxide evolved host using directed host cell evolution. Here we demonstrate that this host has heritable resistance to hydrogen peroxide over many generations, displays enhanced antioxidant capacity through the upregulation of several, diverse antioxidant defense genes such as those involved in glutathione synthesis and turnover, and has improved glutathione content. Additionally, we show that this host has significantly improved transfection recovery times, improved growth and viability properties in a fed-batch production process, and elevated expression of two industrially relevant difficult to express bispecific antibodies compared to unevolved CHO control host cells. These findings demonstrate that host cell evolution represents a powerful methodology for improving specific host cell characteristics that can positively impact the expression of difficult to express biotherapeutics.
中国仓鼠卵巢（CHO）细胞生产双特异性抗体的产量往往低于单克隆抗体。最近，研究表明活性氧会对抗体产生产生负面影响。相比之下，增强细胞抗氧化能力的策略似乎有利于重组蛋白表达。有鉴于此，我们使用定向宿主细胞进化生成了一种新型的过氧化氢进化宿主。在这里，我们证明该宿主在多代中对过氧化氢具有遗传性抗性，通过上调几种不同的抗氧化防御基因（如参与谷胱甘肽合成和周转的基因）显示出增强的抗氧化能力，并且具有改善的谷胱甘肽含量。此外，我们还表明，与未经进化的 CHO 对照宿主细胞相比，该宿主在转染回收时间、补料分批生产过程中的生长和活力特性以及两种工业相关的难以表达的双特异性抗体的表达方面均有显著提高。这些发现表明，宿主细胞进化是一种强大的方法，可以改善特定宿主细胞特性，从而积极影响难以表达的生物治疗药物的表达。
