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• W2336065216 abstract "Chinese hamster ovary cells (CHO) are widely used in the production of glycosylated therapeutic proteins such as antibodies. During expansion and maintenance, CHO cell lines are prone to production instability, which may be caused by promoter silencing, loss of transgene copies, or post-transcriptional effects. Silencing of recombinant genes may be accompanied by DNA methylation and histone modification. Previously, we demonstrated that cytosine methylation of the human cytomegalovirus major immediate early promoter/enhancer (hCMV-MIE) can be used to predict instability of antibody-producing CHO cell lines. However, the high rate of false prediction motivates the search for further markers of stable promoter activity. To this end, we correlated a variety of histone modifications in the vicinity of hCMV-MIE with production stability over time. Our results suggest that acetylation of histone H3 (H3ac) is a more effective indicator of production stability than DNA methylation. Selecting cell lines with highest CMV promoter H3ac levels enriches stable expressors and improves the average stability of production cell lines. For histone H3 acetylation measurement we employed a method based on chromatin immunoprecipitation (ChIP). In its current form, the method is suitable to evaluate 10-20 cell lines within a few days. We propose to determine H3 acetylation once the number of candidate cell lines has been narrowed based on productivity and product quality. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:776-786, 2016." @default.
• W2336065216 created "2016-06-24" @default.
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• W2336065216 date "2016-04-28" @default.
• W2336065216 modified "2023-09-26" @default.
• W2336065216 title "High levels of histone H3 acetylation at the CMV promoter are predictive of stable expression in Chinese hamster ovary cells" @default.
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• W2336065216 doi "https://doi.org/10.1002/btpr.2271" @default.
• W2336065216 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/27073121" @default.
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