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Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells

机译：评估产生mAb的CHO细胞中不同含UCOE的载体组合的表达特征和稳定性

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BackgroundAs the demand for monoclonal antibodies (mAb) increases, more efficient expression methods are required for their manufacturing process. Transcriptional gene silencing is a common phenomenon in recombinant cell lines which leads to expression reduction and instability. There are reports on improved antibody expression in ubiquitous chromatin opening element (UCOE) containing both heavy and light chain gene constructs. Here we investigate the impact of having these elements as part of the light chain, heavy chain or both genes during cell line development. In this regard, non-UCOE and UCOE vectors were constructed and stable Chinese hamster ovary (CHO) cell pools were generated by different vector combinations. ResultsExpression analysis revealed that all UCOE cell pools had higher antibody yields compared to non-UCOE cells, Moreover the most optimal expression was obtained by cells containing just the UCOE on heavy chain. In terms of stability, it was shown that the high level of expression was kept consistence for more than four months in these cells whereas the expression titers were reduced in the other UCOE pools. ConclusionsIn conclusion, UCOE significantly enhanced the level and stability of antibody expression and the use of this element with heavy chain provided more stable cell lines with higher production level.
机译：背景技术随着对单克隆抗体（mAb）的需求增加，其生产过程需要更有效的表达方法。转录基因沉默是重组细胞系中的常见现象，其导致表达降低和不稳定。有报道称，在含有重链和轻链基因构建体的普遍存在的染色质开放元件（UCOE）中，抗体的表达得到改善。在这里，我们研究了在细胞系发育过程中将这些元素作为轻链，重链或两个基因的一部分的影响。在这方面，构建了非UCOE和UCOE载体，并通过不同的载体组合产生了稳定的中国仓鼠卵巢（CHO）细胞库。结果表达分析表明，与非UCOE细胞相比，所有UCOE细胞库均具有更高的抗体产量，而且，仅在重链上仅包含UCOE的细胞获得了最佳表达。在稳定性方面，已表明在这些细胞中高水平的表达保持一致超过四个月，而在其他UCOE库中表达滴度降低。结论综上所述，UCOE显着提高了抗体表达的水平和稳定性，并且该元素与重链一起使用提供了更稳定的细胞系，具有更高的生产水平。

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