Transcriptome analysis of a CHO cell line expressing a recombinant therapeutic protein treated with inducers of protein expression

Journal of Biotechnology
Dina Fomina-YadlinJeffrey T McGrew

Abstract

The search for specific productivity (qP) determinants in Chinese hamster ovary (CHO) cells has been the focus of the biopharmaceutical cell line engineering efforts aimed at creating "super-producer" cell lines. In this study, we evaluated the impact of small-molecule inducers and temperature shift on recombinant protein production, and used transcriptomic analysis to define gene-phenotype correlations for qP in our biological system. Next-generation RNA Sequencing (RNA-Seq) analysis revealed that each individual inducer (caffeine, hexamethylene bisacetamide (HMBA) and sodium butyrate (NaBu)) or a combination treatment had a distinct impact on the gene expression program of the RANK-Fc cell line. Temperature shift to 31 °C impacted inducer action with respect to transcriptional changes and phenotypic cell line parameters. We showed that inducer treatment was able to increase expression level of the Fc- fusion mRNA and the selectable marker mRNA from 16% up to 45% of total mRNA in the cell. We further demonstrated that qP exhibited a strong positive linear correlation to transcript levels of both the RANK-Fc fusion protein and the dihydrofolate reductase (DHFR) selectable marker. In fact, these were 2 out of 7 transcripts with ...Continue Reading

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Citations

Nov 3, 2015·Biotechnology Advances·Simon FischerKerstin Otte
Jan 17, 2017·Scientific Reports·Thomas Beuchert KallehaugeNathan E Lewis
Feb 23, 2020·Biotechnology Letters·Berta Capella RocaMartin Clynes
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Sep 21, 2021·Biotechnology Progress·Markus SchulzeDirk E Martens

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