A multi-omics analysis of recombinant protein production in Hek293 cells

Dietmair, Stefanie, Hodson, Mark P., Quek, Lake-Ee, Timmins, Nicholas E., Gray, Peter and Nielsen, Lars K. (2012) A multi-omics analysis of recombinant protein production in Hek293 cells. Plos One, 7 8: e43394.1-e43394.15.


Author Dietmair, Stefanie
Hodson, Mark P.
Quek, Lake-Ee
Timmins, Nicholas E.
Gray, Peter
Nielsen, Lars K.
Title A multi-omics analysis of recombinant protein production in Hek293 cells
Journal name Plos One  (ERA 2012 Listed)    (ERA 2010 Rank A)   Check publisher's open access policy
Publication date 2012-08
Sub-type Article
DOI 10.1371/journal.pone.0043394
Volume number 7
Issue number 8
ISSN 1932-6203
Start page e43394.1
End page e43394.15
Total pages 15
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Collection year 2013
Language eng
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.
Keyword Transient gene-expression
Mammalian-cells
Pyruvate-carboxylase
Molecule-beta
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Article number e43394

Document type: Journal Article
Sub-type: Article
Collections: Official 2013 Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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