Microarray and proteomics expression profiling identifies several candidates, including the valosin‐containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines
Identifieur interne : 001916 ( Main/Exploration ); précédent : 001915; suivant : 001917Microarray and proteomics expression profiling identifies several candidates, including the valosin‐containing protein (VCP), involved in regulating high cellular growth rate in production CHO cell lines
Auteurs : Padraig Doolan [Irlande (pays)] ; Paula Meleady ; Niall Barron ; Michael Henry ; Ross Gallagher ; Patrick Gammell [Irlande (pays)] ; Mark Melville [États-Unis] ; Martin Sinacore [États-Unis] ; Kevin Mccarthy [États-Unis] ; Mark Leonard [États-Unis] ; Timothy Charlebois [États-Unis] ; Martin ClynesSource :
- Biotechnology and Bioengineering [ 0006-3592 ] ; 2010-05-01.
Abstract
A high rate of cell growth (µ) leading to rapid accumulation of viable biomass is a desirable phenotype during scale up operations and the early stages of production cultures. In order to identify genes and proteins that contribute to higher growth rates in Chinese hamster ovary (CHO) cells, a combined approach using microarray and proteomic expression profiling analysis was carried out on two matched pairs of CHO production cell lines that displayed either fast or slow growth rates. Statistical analysis of the microarray and proteomic data separately resulted in the identification of 118 gene transcripts and 58 proteins that were differentially expressed between the fast‐ and slow‐growing cells. Overlap comparison of both datasets identified a priority list of 21 candidates associated with a high growth rate phenotype in CHO. Functional analysis (by siRNA) of five of these candidates identified the valosin‐containing protein (VCP) as having a substantial impact on CHO cell growth and viability. Knockdown of HSPB1 and ENO1 also had an effect on cell growth (negative and positive, respectively). Further functional validation in CHO using both gene knockdown (siRNA) and overexpression (cDNA) confirmed that altered VCP expression impacted CHO cell proliferation, indicating that VCP and other genes and proteins identified here may play an important role in the regulation of CHO cell growth during log phase culture and are potential candidates for CHO cell line engineering strategies. Biotechnol. Bioeng. 2010; 106: 42–56. © 2010 Wiley Periodicals, Inc.
Url:
DOI: 10.1002/bit.22670
Affiliations:
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type="main">Biotechnology and Bioengineering</title><title level="j" type="alt">BIOTECHNOLOGY AND BIOENGINEERING</title><idno type="ISSN">0006-3592</idno><idno type="eISSN">1097-0290</idno><imprint><biblScope unit="vol">106</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="42">42</biblScope><biblScope unit="page" to="56">56</biblScope><biblScope unit="page-count">15</biblScope><publisher>Wiley Subscription Services, Inc., A Wiley Company</publisher><pubPlace>Hoboken</pubPlace><date type="published" when="2010-05-01">2010-05-01</date></imprint><idno type="ISSN">0006-3592</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0006-3592</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A high rate of cell growth (µ) leading to rapid accumulation of viable biomass is a desirable phenotype during scale up operations and the early stages of production cultures. 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Further functional validation in CHO using both gene knockdown (siRNA) and overexpression (cDNA) confirmed that altered VCP expression impacted CHO cell proliferation, indicating that VCP and other genes and proteins identified here may play an important role in the regulation of CHO cell growth during log phase culture and are potential candidates for CHO cell line engineering strategies. Biotechnol. Bioeng. 2010; 106: 42–56. © 2010 Wiley Periodicals, Inc.</div></front></TEI><affiliations><list><country><li>Irlande (pays)</li><li>États-Unis</li></country><region><li>Massachusetts</li></region></list><tree><noCountry><name sortKey="Barron, Niall" sort="Barron, Niall" uniqKey="Barron N" first="Niall" last="Barron">Niall Barron</name><name sortKey="Clynes, Martin" sort="Clynes, Martin" uniqKey="Clynes M" first="Martin" last="Clynes">Martin Clynes</name><name sortKey="Gallagher, Ross" sort="Gallagher, Ross" uniqKey="Gallagher R" first="Ross" last="Gallagher">Ross Gallagher</name><name sortKey="Henry, Michael" sort="Henry, Michael" uniqKey="Henry M" first="Michael" last="Henry">Michael Henry</name><name sortKey="Meleady, Paula" sort="Meleady, Paula" uniqKey="Meleady P" first="Paula" last="Meleady">Paula Meleady</name></noCountry><country name="Irlande (pays)"><noRegion><name sortKey="Doolan, Padraig" sort="Doolan, Padraig" uniqKey="Doolan P" first="Padraig" last="Doolan">Padraig Doolan</name></noRegion><name sortKey="Gammell, Patrick" sort="Gammell, Patrick" uniqKey="Gammell P" first="Patrick" last="Gammell">Patrick Gammell</name></country><country name="États-Unis"><region name="Massachusetts"><name sortKey="Melville, Mark" sort="Melville, Mark" uniqKey="Melville M" first="Mark" last="Melville">Mark Melville</name></region><name sortKey="Charlebois, Timothy" sort="Charlebois, Timothy" uniqKey="Charlebois T" first="Timothy" last="Charlebois">Timothy Charlebois</name><name sortKey="Leonard, Mark" sort="Leonard, Mark" uniqKey="Leonard M" first="Mark" last="Leonard">Mark Leonard</name><name sortKey="Mccarthy, Kevin" sort="Mccarthy, Kevin" uniqKey="Mccarthy K" first="Kevin" last="Mccarthy">Kevin Mccarthy</name><name sortKey="Sinacore, Martin" sort="Sinacore, Martin" uniqKey="Sinacore M" first="Martin" last="Sinacore">Martin Sinacore</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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