Dietmair 2012 PLoS One

From Bioblast
Publications in the MiPMap
Dietmair S, Hodson MP, Quek LE, Timmins NE, Gray P, Nielsen LK (2012) A multi-omics analysis of recombinant protein production in Hek293 cells. PLoS One 7(8): e43394.

Β» PMID: 22937046 Open Access

Dietmair S, Hodson MP, Quek LE, Timmins NE, Gray P, Nielsen LK (2012) PLoS One

Abstract: Hek293 cells are the predominant hosts for transient expression of recombinant proteins and are used for stable expression of proteins where post-translational modifications performed by CHO cells are inadequate. Nevertheless, there is little information available on the key cellular features underpinning recombinant protein production in Hek293 cells. To improve our understanding of recombinant protein production in Hek293 cells and identify targets for the engineering of an improved host cell line, we have compared a stable, recombinant protein producing Hek293 cell line and its parental cell line using a combination of transcriptomics, metabolomics and fluxomics. Producer cultures consumed less glucose than non-producer cultures while achieving the same growth rate, despite the additional burden of recombinant protein production. Surprisingly, there was no indication that producer cultures compensated for the reduction in glycolytic energy by increasing the efficiency of glucose utilization or increasing glutamine consumption. In contrast, glutamine consumption was lower and the majority of genes involved in oxidative phosphorylation were downregulated in producer cultures. We observed an overall downregulation of a large number of genes associated with broad cellular functions (e.g., cell growth and proliferation) in producer cultures, and therefore speculate that a broad adaptation of the cellular network freed up resources for recombinant protein production while maintaining the same growth rate. Increased abundance of genes associated with endoplasmic reticulum stress indicated a possible bottleneck at the point of protein folding and assembly.


Labels: MiParea: Respiration 


Organism: Human  Tissue;cell: HEK  Preparation: Intact cells 




Pyruvate carboxylase 

The cell dry weight is 514 pg/cell, with 360.5, 86.0, 36.0 and 31.5 pg/cell of protein, lipid, carbohydrate and RNA+DNA, respectively.

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