Glycodelete engineering of mammalian cells simplifies N-glycosylation of recombinant proteins
Open accessAbstract
Heterogeneity in the N-glycans on therapeutic proteins causes difficulties for protein purification and process reproducibility and can lead to variable therapeutic efficacy. This heterogeneity arises from the multistep process of mammalian complex-type N-glycan synthesis. Here we report a glycoengineering strategy--which we call GlycoDelete--that shortens the Golgi N-glycosylation pathway in mammalian cells. This shortening results in the expression of proteins with small, sialylated trisaccharide N-glycans and reduced complexity compared to native mammalian cell glycoproteins. GlycoDelete engineering does not interfere with the functioning of N-glycans in protein folding, and the physiology of cells modified by GlycoDelete is similar to that of wild-type cells. A therapeutic human IgG expressed in GlycoDelete cells had properties, such as reduced initial clearance, that might be beneficial when the therapeutic goal is antigen neutralization. This strategy for reducing N-glycan heterogeneity on mammalian proteins could lead to more consistent performance of therapeutic proteins and modulation of biopharmaceutical functions.
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Using the GlycoDelete expression system for transient anti-CD-20 production, recombinant proteins had very short sialylated N-glycans with high homogeneity on the Fc domain and reduced affinity to FcγRs compared to proteins expressed in wild-type 293S.
“Using this expression system for transient anti-CD-20 production, recombinant proteins had very short sialylated N-glycans with high homogeneity on the Fc domain and reduced affinity to FcγRs compared to proteins expressed in wild-type 293S.”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Meuris et al., 2014
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Experimental — accuracy not yet measuredGlycoDelete reduces glycan heterogeneity by expressing a Golgi-targeted endo-β-N-acetylglucosaminidase in HEK 293SGnTI(−) cells to produce GlcNAc N-glycan stumps modified by galactosyltransferases and sialyltransferases to yield small sialylated trisaccharides.
“GlycoDelete is an elegant glycoengineering strategy worth mentioning, which reduces glycan heterogeneity by expressing a Golgi-targeted endo-β-N-acetylglucosaminidase in HEK 293SGnTI(−) cells to produce GlcNAc N-glycan ‘stumps’ modified by galactosyltransferases and sialyltransferases to yield small sialylated trisaccharides [108].”
From Glycoengineering Chinese hamster ovary cells: a short history (2021) · cites this paper as [108]
A technique called GlycoDelete engineering was recently developed in HEK293S cells.
“Furthermore, to improve N-glycan homogeneity on therapeutic proteins, a technique called “GlycoDelete” engineering was also recently developed in HEK293S cells (Meuris et al., 2014).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Meuris et al., 2014
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- Authors
- Meuris, L., Santens, F., Elson, G., Festjens, N., Boone, M., Dos Santos, A., Devos, S., Rousseau, F., Plets, E., Houthuys, E., Malinge, P., Magistrelli, G., Cons, L., Chatel, L., Devreese, B., & Callewaert, N.
- Affiliations
- Vlaams Instituut voor Biotechnologie (BE)
- Light Chain Bioscience (Switzerland) (CH)
- Ghent University (BE)
- Corresponding author
- Callewaert, Nico
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2014-04-20
- Volume
- 32
- Issue
- 5
- Pages
- 485-489
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC7039703
- OpenAlex
- W1965443132
- Cited by
- 141
- References
- 31
- Citations by year
- 2026: 2 · 2025: 6 · 2024: 5 · 2023: 10 · 2022: 8 · 2021: 12 · 2020: 16 · 2019: 7 · 2018: 22 · 2017: 16 · 2016: 15 · 2015: 16 · 2014: 6
- FWCI
- 7.25
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Ghent University Academic Bibliography (Ghent University) (submittedVersion)
- Funders
- Fonds Wetenschappelijk Onderzoek (G.0.541.08.N.10)
- Universiteit Gent
- Vlaamse regering
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Transgenic Plants and Applications
- Keywords
- N-glycosylation, glycoengineering, recombinant protein expression, protein clearance, glycoprotein folding, biopharmaceuticals
- MeSH terms
- Animals, Glycosylation, Humans, Polysaccharides, Recombinant Proteins, Protein Engineering, Mice
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