Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses
N-linked glycosylation affects the potency, safety, immunogenicity, and pharmacokinetic clearance of several therapeutic proteins including monoclonal antibodies. A robust control strategy is needed to dial in appropriate glycosylation profile during the course of cell culture processes accurately....
| Main Authors: | , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
Elsevier BV
2020
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| Online Access: | https://hdl.handle.net/1721.1/125974 |
| _version_ | 1826205680123510784 |
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| author | Sumit, Madhuresh Dolatshahi, Sepideh Chu, An-Hsiang Adam Cote, Kaffa Scarcelli, John J. Marshall, Jeffrey K. Cornell, Richard J. Weiss, Ron Lauffenburger, Douglas A. Mulukutla, Bhanu Chandra Figueroa, Bruno |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Sumit, Madhuresh Dolatshahi, Sepideh Chu, An-Hsiang Adam Cote, Kaffa Scarcelli, John J. Marshall, Jeffrey K. Cornell, Richard J. Weiss, Ron Lauffenburger, Douglas A. Mulukutla, Bhanu Chandra Figueroa, Bruno |
| author_sort | Sumit, Madhuresh |
| collection | MIT |
| description | N-linked glycosylation affects the potency, safety, immunogenicity, and pharmacokinetic clearance of several therapeutic proteins including monoclonal antibodies. A robust control strategy is needed to dial in appropriate glycosylation profile during the course of cell culture processes accurately. However, N-glycosylation dynamics remains insufficiently understood owing to the lack of integrative analyses of factors that influence the dynamics, including sugar nucleotide donors, glycosyltransferases, and glycosidases. Here, an integrative approach involving multi-dimensional omics analyses was employed to dissect the temporal dynamics of glycoforms produced during fed-batch cultures of CHO cells. Several pathways including glycolysis, tricarboxylic citric acid cycle, and nucleotide biosynthesis exhibited temporal dynamics over the cell culture period. The steps involving galactose and sialic acid addition were determined as temporal bottlenecks. Our results show that galactose, and not manganese, is able to mitigate the temporal bottleneck, despite both being known effectors of galactosylation. Furthermore, sialylation is limited by the galactosylated precursors and autoregulation of cytidine monophosphate-sialic acid biosynthesis. |
| first_indexed | 2024-09-23T13:16:55Z |
| format | Article |
| id | mit-1721.1/125974 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:16:55Z |
| publishDate | 2020 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | mit-1721.1/1259742022-09-28T13:09:21Z Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses Sumit, Madhuresh Dolatshahi, Sepideh Chu, An-Hsiang Adam Cote, Kaffa Scarcelli, John J. Marshall, Jeffrey K. Cornell, Richard J. Weiss, Ron Lauffenburger, Douglas A. Mulukutla, Bhanu Chandra Figueroa, Bruno Massachusetts Institute of Technology. Department of Biological Engineering N-linked glycosylation affects the potency, safety, immunogenicity, and pharmacokinetic clearance of several therapeutic proteins including monoclonal antibodies. A robust control strategy is needed to dial in appropriate glycosylation profile during the course of cell culture processes accurately. However, N-glycosylation dynamics remains insufficiently understood owing to the lack of integrative analyses of factors that influence the dynamics, including sugar nucleotide donors, glycosyltransferases, and glycosidases. Here, an integrative approach involving multi-dimensional omics analyses was employed to dissect the temporal dynamics of glycoforms produced during fed-batch cultures of CHO cells. Several pathways including glycolysis, tricarboxylic citric acid cycle, and nucleotide biosynthesis exhibited temporal dynamics over the cell culture period. The steps involving galactose and sialic acid addition were determined as temporal bottlenecks. Our results show that galactose, and not manganese, is able to mitigate the temporal bottleneck, despite both being known effectors of galactosylation. Furthermore, sialylation is limited by the galactosylated precursors and autoregulation of cytidine monophosphate-sialic acid biosynthesis. 2020-06-24T16:02:54Z 2020-06-24T16:02:54Z 2019-02 2018-11 2020-03-12T17:00:58Z Article http://purl.org/eprint/type/JournalArticle 2589-0042 https://hdl.handle.net/1721.1/125974 Sumit, Madhuresh et al. "Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses." iScience 12 (February 2019): P102-120 © 2019 The Authors en http://dx.doi.org/10.1016/j.isci.2019.01.006 iScience Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier BV Elsevier |
| spellingShingle | Sumit, Madhuresh Dolatshahi, Sepideh Chu, An-Hsiang Adam Cote, Kaffa Scarcelli, John J. Marshall, Jeffrey K. Cornell, Richard J. Weiss, Ron Lauffenburger, Douglas A. Mulukutla, Bhanu Chandra Figueroa, Bruno Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses |
| title | Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses |
| title_full | Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses |
| title_fullStr | Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses |
| title_full_unstemmed | Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses |
| title_short | Dissecting N-Glycosylation Dynamics in Chinese Hamster Ovary Cells Fed-batch Cultures using Time Course Omics Analyses |
| title_sort | dissecting n glycosylation dynamics in chinese hamster ovary cells fed batch cultures using time course omics analyses |
| url | https://hdl.handle.net/1721.1/125974 |
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