Type

Journal Article

Authors

Martin Clynes
Timothy Charlebois
Mark Leonard
Linda Francullo
Kevin McCarthy
Paula Meleady
Martin Sinacore
Patrick Gammell
Mark Melville
Padraig Doolan

Subjects

Biochemistry

Topics
bmp2 protein human bone morphogenetic protein 2 animals transcription genetic recombinant proteins transforming growth factor beta rna messenger genetics cricetulus transfection gene expression profiling cho cells base sequence bone morphogenetic proteins dna primers furin cricetinae humans

Transcriptional profiling of gene expression changes in a PACE-transfected CHO DUKX cell line secreting high levels of rhBMP-2. (2007)

Abstract Chinese hamster ovary (CHO) cells are widely used in the biopharmaceutical industry for the production of recombinant human proteins including complex polypeptides such as recombinant human bone morphogenic protein 2 (rhBMP-2). Large-scale manufacture of rhBMP-2 has associated production difficulties resulting from incomplete processing of the recombinant human protein due to insufficient endogenous levels of the paired basic amino acid cleaving enzyme (PACE) in CHO. In order to resolve this issue, CHO DUKX cells expressing rhBMP-2 were transfected with the soluble version of human PACE (PACEsol) resulting in improved amino-terminal homogeneity and a fourfold increase in rhBMP-2 productivity. In this article, we present a microarray expression profile analysis comparing the parental lineage to the higher producing subclone co-expressing PACEsol using a proprietary CHO-specific microarray. Using this technology we observed 1,076 significantly different genes in the high-productivity cells co-expressing PACEsol. Following further analysis of the differentially expressed genes, the Unfolded Protein Response (UPR) component of the endoplasmic reticulum stress response pathway was identified as a key candidate for effecting increased productivity in this cell system. Several additional ER- and Golgi-localised proteins were identified which may also contribute to this effect. The results presented here support the use of large-scale microarray expression profiling as a viable and valuable route towards understanding the behaviour of bioprocess cultures in vitro.
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Full list of authors on original publication

Martin Clynes, Timothy Charlebois, Mark Leonard, Linda Francullo, Kevin McCarthy, Paula Meleady, Martin Sinacore, Patrick Gammell, Mark Melville, Padraig Doolan

Experts in our system

1
Martin Clynes
Dublin City University
Total Publications: 209
 
2
Paula Meleady
Dublin City University
Total Publications: 95
 
3
Padraig Doolan
Dublin City University
Total Publications: 37