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A proteomic study of cMyc improvement of CHO culture

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,...

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Autores principales: Kuystermans, Darrin, Dunn, Michael J, Al-Rubeai, Mohamed
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2859402/
https://www.ncbi.nlm.nih.gov/pubmed/20307306
http://dx.doi.org/10.1186/1472-6750-10-25
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author Kuystermans, Darrin
Dunn, Michael J
Al-Rubeai, Mohamed
author_facet Kuystermans, Darrin
Dunn, Michael J
Al-Rubeai, Mohamed
author_sort Kuystermans, Darrin
collection PubMed
description 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.
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spelling pubmed-28594022010-04-27 A proteomic study of cMyc improvement of CHO culture Kuystermans, Darrin Dunn, Michael J Al-Rubeai, Mohamed BMC Biotechnol Research article 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. BioMed Central 2010-03-22 /pmc/articles/PMC2859402/ /pubmed/20307306 http://dx.doi.org/10.1186/1472-6750-10-25 Text en Copyright ©2010 Kuystermans et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research article
Kuystermans, Darrin
Dunn, Michael J
Al-Rubeai, Mohamed
A proteomic study of cMyc improvement of CHO culture
title A proteomic study of cMyc improvement of CHO culture
title_full A proteomic study of cMyc improvement of CHO culture
title_fullStr A proteomic study of cMyc improvement of CHO culture
title_full_unstemmed A proteomic study of cMyc improvement of CHO culture
title_short A proteomic study of cMyc improvement of CHO culture
title_sort proteomic study of cmyc improvement of cho culture
topic Research article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2859402/
https://www.ncbi.nlm.nih.gov/pubmed/20307306
http://dx.doi.org/10.1186/1472-6750-10-25
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