Nematpour Fatemeh, Mahboudi Fereidoun, Vaziri Behrouz, Khalaj Vahid, Ahmadi Samira, Ahmadi Maryam, Ebadat Saedeh, Davami Fatemeh
Biotechnology Research Center, Pasteur Institute of Iran, Tehran, 1316943551, Iran.
Departments of Medical Biotechnology, Semnan University of Medical Sciences, Semnan, 3519899951, Iran.
BMC Biotechnol. 2017 Feb 22;17(1):18. doi: 10.1186/s12896-017-0330-0.
As 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.
Expression 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.
In 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.
随着单克隆抗体（mAb）需求的增加，其生产过程需要更高效的表达方法。转录基因沉默是重组细胞系中的常见现象，会导致表达降低和不稳定。有报道称，在包含重链和轻链基因构建体的遍在染色质开放元件（UCOE）中，抗体表达有所改善。在此，我们研究了在细胞系开发过程中，将这些元件作为轻链、重链或两个基因的一部分时的影响。为此，构建了非UCOE和UCOE载体，并通过不同的载体组合生成了稳定的中国仓鼠卵巢（CHO）细胞库。
表达分析显示，与非UCOE细胞相比，所有UCOE细胞库的抗体产量更高。此外，仅在重链上含有UCOE的细胞获得了最优化的表达。在稳定性方面，结果表明，这些细胞中的高水平表达在四个多月内保持一致，而其他UCOE细胞库中的表达滴度则有所降低。
总之，UCOE显著提高了抗体表达的水平和稳定性，并且将该元件与重链一起使用可提供更稳定、产量更高的细胞系。
