A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells

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 underp...

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Main Authors:	Dietmair, Stefanie, Hodson, Mark P., Quek, Lake-Ee, Timmins, Nicholas E., Gray, Peter, Nielsen, Lars K.
Format:	Online
Language:	English
Published:	Public Library of Science 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3427347/

id	pubmed-3427347
recordtype	oai_dc
spelling	pubmed-34273472012-08-30 A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells Dietmair, Stefanie Hodson, Mark P. Quek, Lake-Ee Timmins, Nicholas E. Gray, Peter Nielsen, Lars K. Research Article 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. Public Library of Science 2012-08-24 /pmc/articles/PMC3427347/ /pubmed/22937046 http://dx.doi.org/10.1371/journal.pone.0043394 Text en © 2012 Dietmair et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Dietmair, Stefanie Hodson, Mark P. Quek, Lake-Ee Timmins, Nicholas E. Gray, Peter Nielsen, Lars K.
spellingShingle	Dietmair, Stefanie Hodson, Mark P. Quek, Lake-Ee Timmins, Nicholas E. Gray, Peter Nielsen, Lars K. A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells
author_facet	Dietmair, Stefanie Hodson, Mark P. Quek, Lake-Ee Timmins, Nicholas E. Gray, Peter Nielsen, Lars K.
author_sort	Dietmair, Stefanie
title	A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells
title_short	A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells
title_full	A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells
title_fullStr	A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells
title_full_unstemmed	A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells
title_sort	multi-omics analysis of recombinant protein production in hek293 cells
description	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.
publisher	Public Library of Science
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3427347/
_version_	1611552157942677504

A Multi-Omics Analysis of Recombinant Protein Production in Hek293 Cells

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