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|Title:||LC-MS-based metabolic characterization of high monoclonal antibody-producing Chinese hamster ovary cells||Authors:||Chong, W.P.K.
|Keywords:||Chinese hamster ovary (CHO)
Liquid chromatography-mass spectrometry (LC-MS)
Monoclonal antibody (mAb)
|Issue Date:||Dec-2012||Citation:||Chong, W.P.K., Thng, S.H., Hiu, A.P., Lee, D.-Y., Chan, E.C.Y., Ho, Y.S. (2012-12). LC-MS-based metabolic characterization of high monoclonal antibody-producing Chinese hamster ovary cells. Biotechnology and Bioengineering 109 (12) : 3103-3111. ScholarBank@NUS Repository. https://doi.org/10.1002/bit.24580||Abstract:||The selection of suitable mammalian cell lines with high specific productivities is a crucial aspect of large-scale recombinant protein production. This study utilizes a metabolomics approach to elucidate the key characteristics of Chinese hamster ovary (CHO) cells with high monoclonal antibody productivities (qmAb). Liquid chromatography-mass spectrometry (LC-MS)-based intracellular metabolite profiles of eight single cell clones with high and low qmAb were obtained at the mid-exponential phase during shake flask batch cultures. Orthogonal projection to latent structures discriminant analysis (OPLS-DA) subsequently revealed key differences between the high and low qmAb clones, as indicated by the variable importance for projection (VIP) scores. The mass peaks were further examined for their potential association with qmAb across all clones using Pearson's correlation analysis. Lastly, the identities of metabolites with high VIP and correlation scores were confirmed by comparison with standards through LC-MS-MS. A total of seven metabolites were identified-NADH, FAD, reduced and oxidized glutathione, and three activated sugar precursors. These metabolites are involved in key cellular pathways of citric acid cycle, oxidative phosphorylation, glutathione metabolism, and protein glycosylation. To our knowledge, this is the first study to identify metabolites that are associated closely with qmAb. The results suggest that the high producers had elevated levels of specific metabolites to better regulate their redox status. This is likely to facilitate the generation of energy and activated sugar precursors to meet the demands of producing more glycosylated recombinant monoclonal antibodies. © 2012 Wiley Periodicals, Inc.||Source Title:||Biotechnology and Bioengineering||URI:||http://scholarbank.nus.edu.sg/handle/10635/89321||ISSN:||00063592||DOI:||10.1002/bit.24580|
|Appears in Collections:||Staff Publications|
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