Cited by 141Viral Infectious Diseases and Gene Expression in InsectsGlycosylation and Glycoproteins ResearchTransgenic Plants and Applications

Glycodelete engineering of mammalian cells simplifies N-glycosylation of recombinant proteins

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Meuris, Leander, Santens, Francis, Elson, Greg, Festjens, Nele, Boone, Morgane, Dos Santos, Anaëlle, Devos, Simon, Rousseau, François +8 more·2014·Nature Biotechnology

Abstract

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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  • GlycoDelete 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
Venue
Nature Biotechnology
Published
2014-04-20
Volume
32
Issue
5
Pages
485-489
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
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
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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