Suppr超能文献

通过优化表达载体和翻译效率提高抗 IL-2Rα 单克隆抗体在 CHO 细胞中的表达。

Improving the expression of anti-IL-2Rα monoclonal antibody in the CHO cells through optimization of the expression vector and translation efficiency.

机构信息

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

出版信息

J Biotechnol. 2020 Dec 20;324:112-120. doi: 10.1016/j.jbiotec.2020.09.006. Epub 2020 Sep 29.

Abstract

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.

摘要

单克隆抗体(mAbs)的需求不断增长,这就需要开发新的、高效的生产方法。广泛存在的染色质开放元件(UCOEs)等调控元件被用于提高中国仓鼠卵巢(CHO)细胞中 mAb 的表达。SINEUPs 是一类长非编码 RNA,可以提高部分重叠 mRNA 的翻译效率。这两种元件的结合可能会导致 mAb 产量的提高。因此,本研究旨在探讨 SINEUPs 和 A2UCOE 对 CHO-K1 细胞中 IgG1 表达的影响。为此,构建了 mAb、mAb-SINEUP 和 mAb-UCOE 载体后,通过组合上述载体生成了四个稳定的细胞池。根据表达分析,mAb-SINEUP 和 mAb-UCOE 细胞池的抗体产量高于 mAb 细胞。此外,同时具有 SINEUP 和 UCOE 元件的细胞提供了最佳表达。这些细胞中持续的 mAb 表达超过 2 个月,而 mAb 池中的表达则下降。SINEUP 和 UCOE 对稳定的 mAb 表达有积极影响。可以得出结论,SINEUP 和 UCOE 分别提高了抗体的稳定性和表达水平,它们的结合提高了 CHO 细胞中 mAb 的产量。

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验