Abstract

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.

Highlights

  • As the demand for monoclonal antibodies increases, more efficient expression methods are required for their manufacturing process

  • Analysis between ubiquitous chromatin opening element (UCOE) pools showed that the expression level driven by Chinese hamster ovary (CHO)-UHL was greater than other UCOE cell pools

  • In line with previous studies, these results indicate that the inclusion of UCOE on antibody chains provides a substantial improvement in CHO cell productivity [16, 21]

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Summary

Introduction

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. 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. It has been reported that the use of cis-acting epigenetic regulatory elements such as locus control regions (LCRs), matrix attachment regions (MARs) and ubiquitous chromatin opening elements (UCOEs) can protect transgenes from such

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