Improving the expression of anti-IL-2Rα monoclonal antibody in the CHO cells through optimization of the expression vector and translation efficiency
Improving the expression of anti-IL-2Rα monoclonal antibody in the CHO cells through optimization of the expression vector and translation efficiency
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DOI：
10.1016/j.jbiotec.2020.09.006
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发表时间：
2020-12-20
影响因子：
4.1
通讯作者：
Rahimpour, Azam
中科院分区：
文献类型：
--
作者：
Hoseinpoor, Reyhaneh;Kazemi, Bahram;Rahimpour, Azam
对单克隆抗体 (mAb) 的需求不断增长，需要开发新颖且高效的生产方法。普遍存在的染色质开放元件 (UCOE) 等调节元件已被用于改善中国仓鼠卵巢 (CHO) 细胞中的 mAb 表达。 SINEUP是一类长非编码RNA，可以改善部分重叠mRNA的翻译。这两种元素的组合可能会导致单克隆抗体的更高产量。因此，本研究旨在探讨 SINEUP 和 A2UCOE 对 CHO-K1 细胞中 IgG1 表达的影响。因此，在构建mAb、mAb-SINEUP和mAb-UCOE载体后，通过组合上述载体产生了四个稳定细胞池。根据表达分析，与 mAb 细胞相比，mAb-SINEUP 和 mAb-UCOE 细胞库中的抗体产量更高。此外，同时具有SINEUP和UCOE元件的细胞提供了最好的表达。在这些细胞中观察到持续的 mAb 表达超过 2 个月，而 mAb 池中的表达却有所下降。 SINEUP 和 UCOE 对稳定的 mAb 表达产生积极影响。可以得出结论，SINEUP 和 UCOE 分别增强了抗体稳定性和表达水平，它们的组合提高了 CHO 细胞中 mAb 的产量。
The growing need for monoclonal antibodies (mAbs) necessitates the development of novel and efficient production approaches. Regulatory elements like ubiquitous chromatin-opening elements (UCOEs) have been employed for improvement of the mAb expression in the Chinese hamster ovary (CHO) cells. SINEUPs are a class of long non-coding RNAs, which can improve the translation of partly overlapping mRNAs. A combination of these two elements might lead to higher production of mAbs. Therefore, the current study was conducted to investigate the effects of SINEUPs and A2UCOE on the expression of an IgG1 in the CHO-K1 cells. Hence, after constructing the mAb, mAb-SINEUP, and mAb-UCOE vectors, four stable cell pools were generated through combining the above vectors. According to the expression analysis, antibody yields were higher in the mAb-SINEUP and mAb-UCOE cell pools compared to the mAb cells. In addition, the cells possessing both SINEUP and UCOE elements provided the best expression. Persistent mAb expression was observed for over 2 months in these cells, whilst the expression was decreased in the mAb pool. SINEUP and UCOE positively influenced the stable mAb expression. It can be concluded that the SINEUP and UCOE enhance the antibody stability and expression level separately and their combination improves the mAb production in the CHO cells.