A proteomic study of cMyc improvement of CHO culture

Hdl Handle:
http://hdl.handle.net/10147/239176
Title:
A proteomic study of cMyc improvement of CHO culture
Authors:
Kuystermans, Darrin; Dunn, Michael J; Al-Rubeai, Mohamed
Citation:
BMC Biotechnology. 2010 Mar 22;10(1):25
Issue Date:
22-Mar-2010
URI:
http://dx.doi.org/10.1186/1472-6750-10-25; http://hdl.handle.net/10147/239176
Abstract:
Abstract Background The biopharmaceutical industry requires cell lines to have an optimal proliferation rate and a high integral viable cell number resulting in a maximum volumetric recombinant protein product titre. Nutrient feeding has been shown to boost cell number and productivity in fed-batch culture, but cell line engineering is another route one may take to increase these parameters in the bioreactor. The use of CHO-K1 cells with a c-myc plasmid allowing for over-expressing c-Myc (designated cMycCHO) gives a higher integral viable cell number. In this study the differential protein expression in cMycCHO is investigated using two-dimensional gel electrophoresis (2-DE) followed by image analysis to determine the extent of the effect c-Myc has on the cell and the proteins involved to give the new phenotype. Results Over 100 proteins that were differentially expressed in cMycCHO cells were detected with high statistical confidence, of which 41 were subsequently identified by tandem mass spectrometry (LC-MS/MS). Further analysis revealed proteins involved in a variety of pathways. Some examples of changes in protein expression include: an increase in nucleolin, involved in proliferation and known to aid in stabilising anti-apoptotic protein mRNA levels, the cytoskeleton and mitochondrial morphology (vimentin), protein biosysnthesis (eIF6) and energy metabolism (ATP synthetase), and a decreased regulation of all proteins, indentified, involved in matrix and cell to cell adhesion. Conclusion These results indicate several proteins involved in proliferation and adhesion that could be useful for future approaches to improve proliferation and decrease adhesion of CHO cell lines which are difficult to adapt to suspension culture.
Item Type:
Journal Article

Full metadata record

DC FieldValue Language
dc.contributor.authorKuystermans, Darrin-
dc.contributor.authorDunn, Michael J-
dc.contributor.authorAl-Rubeai, Mohamed-
dc.date.accessioned2012-08-20T11:58:00Z-
dc.date.available2012-08-20T11:58:00Z-
dc.date.issued2010-03-22-
dc.identifier.citationBMC Biotechnology. 2010 Mar 22;10(1):25-
dc.identifier.urihttp://dx.doi.org/10.1186/1472-6750-10-25-
dc.identifier.urihttp://hdl.handle.net/10147/239176-
dc.description.abstractAbstract Background The biopharmaceutical industry requires cell lines to have an optimal proliferation rate and a high integral viable cell number resulting in a maximum volumetric recombinant protein product titre. Nutrient feeding has been shown to boost cell number and productivity in fed-batch culture, but cell line engineering is another route one may take to increase these parameters in the bioreactor. The use of CHO-K1 cells with a c-myc plasmid allowing for over-expressing c-Myc (designated cMycCHO) gives a higher integral viable cell number. In this study the differential protein expression in cMycCHO is investigated using two-dimensional gel electrophoresis (2-DE) followed by image analysis to determine the extent of the effect c-Myc has on the cell and the proteins involved to give the new phenotype. Results Over 100 proteins that were differentially expressed in cMycCHO cells were detected with high statistical confidence, of which 41 were subsequently identified by tandem mass spectrometry (LC-MS/MS). Further analysis revealed proteins involved in a variety of pathways. Some examples of changes in protein expression include: an increase in nucleolin, involved in proliferation and known to aid in stabilising anti-apoptotic protein mRNA levels, the cytoskeleton and mitochondrial morphology (vimentin), protein biosysnthesis (eIF6) and energy metabolism (ATP synthetase), and a decreased regulation of all proteins, indentified, involved in matrix and cell to cell adhesion. Conclusion These results indicate several proteins involved in proliferation and adhesion that could be useful for future approaches to improve proliferation and decrease adhesion of CHO cell lines which are difficult to adapt to suspension culture.-
dc.titleA proteomic study of cMyc improvement of CHO culture-
dc.typeJournal Article-
dc.language.rfc3066en-
dc.rights.holderDarrin Kuystermans et al.; licensee BioMed Central Ltd.-
dc.description.statusPeer Reviewed-
dc.date.updated2012-08-20T11:08:24Z-
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