Analysis of dynamic changes in the proteome of a Bcl-XL overexpressing Chinese hamster ovary cell culture during exponential and stationary phases

Biotechnology Progress
Tyler CarlageWilliam S Hancock

Abstract

Mammalian cell cultures used for biopharmaceutical production undergo various dynamic biological changes over time, including the transition of cells from an exponential growth phase to a stationary phase during cell culture. To better understand the dynamic aspects of cell culture, a quantitative proteomics approach was used to identify dynamic trends in protein expression over the course of a Chinese hamster ovary (CHO) cell culture for the production of a recombinant monoclonal antibody and overexpressing the antiapoptotic gene Bcl-xl. Samples were analyzed using a method incorporating iTRAQ labeling, two-dimensional LC/MS, and linear regression calculations to identify significant dynamic trends in protein abundance. Using this approach, 59 proteins were identified with significant temporal changes in expression. Pathway analysis tools were used to identify a putative network of proteins associated with cell growth and apoptosis. Among the differentially expressed proteins were molecular chaperones and isomerases, such as GRP78 and PDI, and reported cell growth markers MCM2 and MCM5. In addition, two proteins with growth-regulating properties, transglutaminase-2 and clusterin, were identified. These proteins are associated ...Continue Reading

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Citations

Oct 12, 2012·Applied Microbiology and Biotechnology·M Butler, A Meneses-Acosta
Sep 19, 2015·Critical Reviews in Biotechnology·Jennifer DumontRashmi Kshirsagar
Sep 30, 2015·Journal of Proteome Research·Amit KumarMichael J Betenbaugh
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Jul 10, 2021·Current Opinion in Biotechnology·Pritam SinharoyMichael W Handlogten
Sep 7, 2017·Journal of Proteome Research·Kelley M HeffnerMichael J Betenbaugh

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