Small-molecule control of antibody N-glycosylation in engineered mammalian cells
N-linked glycosylation in monoclonal antibodies (mAbs) is crucial for structural and functional properties of mAb therapeutics, including stability, pharmacokinetics, safety and clinical efficacy. The biopharmaceutical industry currently lacks tools to precisely control N-glycosylation levels during...
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| Format: | Article |
| Language: | English |
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Springer Science and Business Media LLC
2020
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| Online Access: | https://hdl.handle.net/1721.1/125977 |
| _version_ | 1826202991376465920 |
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| author | Chang, Michelle Gaidukov, Leonid Jung, Giyoung Tseng, Wen Allen Scarcelli, John J. Cornell, Richard Marshall, Jeffrey K. Lyles, Jonathan L Sakorafas, Paul Chu, An-Hsiang Adam Cote, Kaffa Tzvetkova, Boriana Dolatshahi, Sepideh Sumit, Madhuresh Mulukutla, Bhanu Chandra Lauffenburger, Douglas A Figueroa, Bruno Summers, Nevin M Lu, Timothy K Weiss, Ron |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Chang, Michelle Gaidukov, Leonid Jung, Giyoung Tseng, Wen Allen Scarcelli, John J. Cornell, Richard Marshall, Jeffrey K. Lyles, Jonathan L Sakorafas, Paul Chu, An-Hsiang Adam Cote, Kaffa Tzvetkova, Boriana Dolatshahi, Sepideh Sumit, Madhuresh Mulukutla, Bhanu Chandra Lauffenburger, Douglas A Figueroa, Bruno Summers, Nevin M Lu, Timothy K Weiss, Ron |
| author_sort | Chang, Michelle |
| collection | MIT |
| description | N-linked glycosylation in monoclonal antibodies (mAbs) is crucial for structural and functional properties of mAb therapeutics, including stability, pharmacokinetics, safety and clinical efficacy. The biopharmaceutical industry currently lacks tools to precisely control N-glycosylation levels during mAb production. In this study, we engineered Chinese hamster ovary cells with synthetic genetic circuits to tune N-glycosylation of a stably expressed IgG. We knocked out two key glycosyltransferase genes, α-1,6-fucosyltransferase (FUT8) and β-1,4-galactosyltransferase (β4GALT1), genomically integrated circuits expressing synthetic glycosyltransferase genes under constitutive or inducible promoters and generated antibodies with concurrently desired fucosylation (0–97%) and galactosylation (0–87%) levels. Simultaneous and independent control of FUT8 and β4GALT1 expression was achieved using orthogonal small molecule inducers. Effector function studies confirmed that glycosylation profile changes affected antibody binding to a cell surface receptor. Precise and rational modification of N-glycosylation will allow new recombinant protein therapeutics with tailored in vitro and in vivo effects for various biotechnological and biomedical applications. |
| first_indexed | 2024-09-23T12:29:24Z |
| format | Article |
| id | mit-1721.1/125977 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T12:29:24Z |
| publishDate | 2020 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1259772022-09-28T08:07:53Z Small-molecule control of antibody N-glycosylation in engineered mammalian cells Small-molecule control of antibody N-glycosylation in engineered mammalian cells Chang, Michelle Gaidukov, Leonid Jung, Giyoung Tseng, Wen Allen Scarcelli, John J. Cornell, Richard Marshall, Jeffrey K. Lyles, Jonathan L Sakorafas, Paul Chu, An-Hsiang Adam Cote, Kaffa Tzvetkova, Boriana Dolatshahi, Sepideh Sumit, Madhuresh Mulukutla, Bhanu Chandra Lauffenburger, Douglas A Figueroa, Bruno Summers, Nevin M Lu, Timothy K Weiss, Ron Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Synthetic Biology Center Harvard University--MIT Division of Health Sciences and Technology N-linked glycosylation in monoclonal antibodies (mAbs) is crucial for structural and functional properties of mAb therapeutics, including stability, pharmacokinetics, safety and clinical efficacy. The biopharmaceutical industry currently lacks tools to precisely control N-glycosylation levels during mAb production. In this study, we engineered Chinese hamster ovary cells with synthetic genetic circuits to tune N-glycosylation of a stably expressed IgG. We knocked out two key glycosyltransferase genes, α-1,6-fucosyltransferase (FUT8) and β-1,4-galactosyltransferase (β4GALT1), genomically integrated circuits expressing synthetic glycosyltransferase genes under constitutive or inducible promoters and generated antibodies with concurrently desired fucosylation (0–97%) and galactosylation (0–87%) levels. Simultaneous and independent control of FUT8 and β4GALT1 expression was achieved using orthogonal small molecule inducers. Effector function studies confirmed that glycosylation profile changes affected antibody binding to a cell surface receptor. Precise and rational modification of N-glycosylation will allow new recombinant protein therapeutics with tailored in vitro and in vivo effects for various biotechnological and biomedical applications. 2020-06-25T14:07:15Z 2020-06-25T14:07:15Z 2019-05 2018-08 2020-06-19T13:11:23Z Article http://purl.org/eprint/type/JournalArticle 1552-4450 1552-4469 https://hdl.handle.net/1721.1/125977 Chang, Michelle M. et al. "Small-molecule control of antibody N-glycosylation in engineered mammalian cells." Nature Chemical Biology 15, 7 (May 2019): 730–736 © 2019 The Author(s) en http://dx.doi.org/10.1038/s41589-019-0288-4 Nature Chemical Biology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Springer Science and Business Media LLC Prof. Lauffenberger via Howard Silver |
| spellingShingle | Chang, Michelle Gaidukov, Leonid Jung, Giyoung Tseng, Wen Allen Scarcelli, John J. Cornell, Richard Marshall, Jeffrey K. Lyles, Jonathan L Sakorafas, Paul Chu, An-Hsiang Adam Cote, Kaffa Tzvetkova, Boriana Dolatshahi, Sepideh Sumit, Madhuresh Mulukutla, Bhanu Chandra Lauffenburger, Douglas A Figueroa, Bruno Summers, Nevin M Lu, Timothy K Weiss, Ron Small-molecule control of antibody N-glycosylation in engineered mammalian cells |
| title | Small-molecule control of antibody N-glycosylation in engineered mammalian cells |
| title_full | Small-molecule control of antibody N-glycosylation in engineered mammalian cells |
| title_fullStr | Small-molecule control of antibody N-glycosylation in engineered mammalian cells |
| title_full_unstemmed | Small-molecule control of antibody N-glycosylation in engineered mammalian cells |
| title_short | Small-molecule control of antibody N-glycosylation in engineered mammalian cells |
| title_sort | small molecule control of antibody n glycosylation in engineered mammalian cells |
| url | https://hdl.handle.net/1721.1/125977 |
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