High-Antibody-Producing Chinese Hamster Ovary Cells Up-Regulate Intracellular Protein Transport and Glutathione Synthesis

Chinese hamster ovary (CHO) cells are the preferred production host for therapeutic monoclonal antibodies (mAb) due to their ability to perform post-translational modifications and their successful approval history. The completion of the genome sequence for CHO cells has reignited interest in using...

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Published inJournal of proteome research Vol. 14; no. 2; pp. 609 - 618
Main Authors Orellana, Camila A, Marcellin, Esteban, Schulz, Benjamin L, Nouwens, Amanda S, Gray, Peter P, Nielsen, Lars K
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 06.02.2015
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Abstract Chinese hamster ovary (CHO) cells are the preferred production host for therapeutic monoclonal antibodies (mAb) due to their ability to perform post-translational modifications and their successful approval history. The completion of the genome sequence for CHO cells has reignited interest in using quantitative proteomics to identify markers of good production lines. Here we applied two different proteomic techniques, iTRAQ and SWATH, for the identification of expression differences between a high- and low-antibody-producing CHO cell lines derived from the same transfection. More than 2000 proteins were quantified with 70 of them classified as differentially expressed in both techniques. Two biological processes were identified as differentially regulated by both methods: up-regulation of glutathione biosynthesis and down-regulation of DNA replication. Metabolomic analysis confirmed that the high producing cell line displayed higher intracellular levels of glutathione. SWATH further identified up-regulation of actin filament processes and intracellular transport and down regulation of several growth-related processes. These processes may be important for conferring high mAb production and as such are promising candidates for targeted engineering of high-expression cell lines.
AbstractList Chinese hamster ovary (CHO) cells are the preferred production host for therapeutic monoclonal antibodies (mAb) due to their ability to perform post-translational modifications and their successful approval history. The completion of the genome sequence for CHO cells has reignited interest in using quantitative proteomics to identify markers of good production lines. Here we applied two different proteomic techniques, iTRAQ and SWATH, for the identification of expression differences between a high- and low-antibody-producing CHO cell lines derived from the same transfection. More than 2000 proteins were quantified with 70 of them classified as differentially expressed in both techniques. Two biological processes were identified as differentially regulated by both methods: up-regulation of glutathione biosynthesis and down-regulation of DNA replication. Metabolomic analysis confirmed that the high producing cell line displayed higher intracellular levels of glutathione. SWATH further identified up-regulation of actin filament processes and intracellular transport and down regulation of several growth-related processes. These processes may be important for conferring high mAb production and as such are promising candidates for targeted engineering of high-expression cell lines.
Author Gray, Peter P
Orellana, Camila A
Nouwens, Amanda S
Nielsen, Lars K
Marcellin, Esteban
Schulz, Benjamin L
AuthorAffiliation The University of Queensland
School of Chemistry and Molecular Biosciences
Australian Institute for Bioengineering and Nanotechnology (AIBN)
AuthorAffiliation_xml – name: Australian Institute for Bioengineering and Nanotechnology (AIBN)
– name: School of Chemistry and Molecular Biosciences
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BackLink https://www.ncbi.nlm.nih.gov/pubmed/25495469$$D View this record in MEDLINE/PubMed
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Keywords iTRAQ
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CHO cells
glutathione
quantitative proteomics
monoclonal antibodies
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Snippet Chinese hamster ovary (CHO) cells are the preferred production host for therapeutic monoclonal antibodies (mAb) due to their ability to perform...
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SubjectTerms Animals
Antibodies, Monoclonal - biosynthesis
CHO Cells
Cricetinae
Cricetulus
Female
Glutathione - biosynthesis
Ovary - immunology
Protein Transport
Up-Regulation
Title High-Antibody-Producing Chinese Hamster Ovary Cells Up-Regulate Intracellular Protein Transport and Glutathione Synthesis
URI http://dx.doi.org/10.1021/pr501027c
https://www.ncbi.nlm.nih.gov/pubmed/25495469
https://search.proquest.com/docview/1652458463
Volume 14
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