Small-molecule control of antibody N-glycosylation in engineered mammalian cells
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Description
Accepted version
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Author(s)
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
Alternative Title
Small-molecule control of antibody N-glycosylation in engineered mammalian cells
Date Issued
May 2019
Journal
Nature Chemical Biology
Publisher
Springer Science and Business Media LLC
Citation
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)
Version
Author's final manuscript
Abstract
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.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Synthetic Biology Center
Harvard University--MIT Division of Health Sciences and Technology
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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.
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DOI of Published Version
https://doi.org/10.1038/s41589-019-0288-4
