Stable microRNA expression enhances therapeutic antibody productivity of Chinese hamster ovary cells

MicroRNAs (miRNAs) are short non-coding RNAs that post-transcriptionally regulate the expression of different target genes and, thus, enable engineered gene networks to achieve complex phenotypic changes in mammalian cells. We hypothesized that exploiting this feature of miRNAs could improve therape...

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Bibliographic Details
Published in:Metabolic engineering 2013-11, Vol.20, p.157-166
Main Authors: Strotbek, Michaela, Florin, Lore, Koenitzer, Jennifer, Tolstrup, Anne, Kaufmann, Hitto, Hausser, Angelika, Olayioye, Monilola A.
Format: Article
Language:English
Subjects:
Animals
Antibodies
Antibodies, Monoclonal - biosynthesis
Antibodies, Monoclonal - genetics
Cell line engineering
CHO Cells
Cricetinae
Cricetulus
Fed-batch culture
Functional genomic screen
Genome-Wide Association Study
Humans
Immunoglobulin G - biosynthesis
Immunoglobulin G - genetics
MicroRNAs - biosynthesis
MicroRNAs - genetics
miRNA library
Recombinant therapeutic protein production
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Summary:MicroRNAs (miRNAs) are short non-coding RNAs that post-transcriptionally regulate the expression of different target genes and, thus, enable engineered gene networks to achieve complex phenotypic changes in mammalian cells. We hypothesized that exploiting this feature of miRNAs could improve therapeutic protein production processes by increasing viable cell densities and/or productivity of the mammalian cells used for manufacturing. To identify miRNAs that increase the productivity of producer cells, we performed a genome wide functional miRNA screen by transient transfection of Chinese hamster ovary (CHO) cells stably expressing an IgG1 antibody (CHO-IgG1). Using this approach, we identified nine human miRNAs that improved the productivities not only of the CHO-IgG1 cells but also of CHO cells expressing recombinant human serum albumin (HSA), demonstrating that the miRNAs act in a product-independent manner. We selected two miRNAs (miR-557 and miR-1287) positively impacting the viable cell density and the specific productivity, respectively, and then stably co-expressed them in IgG1 expressing CHO cells. In these cells, higher IgG1 titers were observed in fed-batch cultures whilst product quality was conserved, demonstrating that miRNA-based cell line engineering provides an attractive approach toward the genetic optimization of CHO producer cells for industrial applications. •First genome wide miRNA screen in an IgG1-secreting CHO cell line.•Identification of nine miRNAs enhancing IgG1 production levels.•Improved performance of stable miR-engineered CHO cells in fed-batch cultures.
ISSN:1096-7176
1096-7184
DOI:10.1016/j.ymben.2013.10.005