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Nat Biotechnol. 2014 May;32(5):485-9. doi: 10.1038/nbt.2885. Epub 2014 Apr 20.

GlycoDelete engineering of mammalian cells simplifies N-glycosylation of recombinant proteins.

Author information

1
1] Unit for Medical Biotechnology, Inflammation Research Center (IRC), VIB, Ghent, Belgium. [2] Laboratory for Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium. [3].
2
1] NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland. [2].
3
1] Unit for Medical Biotechnology, Inflammation Research Center (IRC), VIB, Ghent, Belgium. [2] Laboratory for Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium.
4
NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland.
5
Laboratory for Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium.

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.

Comment in

PMID:
24752077
DOI:
10.1038/nbt.2885
[Indexed for MEDLINE]

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