title: GlycoDelete engineering of mammalian cells simplifies N-glycosylation of recombinant proteins.
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.
authors.affiliation: 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].
authors.forename: Leander
authors.initials: L
authors.lastname: Meuris
authors.affiliation: 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].
authors.forename: Francis
authors.initials: F
authors.lastname: Santens
authors.affiliation: 1] NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland. [2].
authors.forename: Greg
authors.initials: G
authors.lastname: Elson
authors.affiliation: 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.
authors.forename: Nele
authors.initials: N
authors.lastname: Festjens
authors.affiliation: 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.
authors.forename: Morgane
authors.initials: M
authors.lastname: Boone
authors.affiliation: NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland.
authors.forename: Anaëlle
authors.initials: A
authors.lastname: Dos Santos
authors.affiliation: Laboratory for Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium.
authors.forename: Simon
authors.initials: S
authors.lastname: Devos
authors.affiliation: NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland.
authors.forename: François
authors.initials: F
authors.lastname: Rousseau
authors.affiliation: 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.
authors.forename: Evelyn
authors.initials: E
authors.lastname: Plets
authors.affiliation: 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.
authors.forename: Erica
authors.initials: E
authors.lastname: Houthuys
authors.affiliation: NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland.
authors.forename: Pauline
authors.initials: P
authors.lastname: Malinge
authors.affiliation: NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Magistrelli
authors.affiliation: NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland.
authors.forename: Laura
authors.initials: L
authors.lastname: Cons
authors.affiliation: NovImmune SA, Plan-Les-Ouates, Geneva, Switzerland.
authors.forename: Laurence
authors.initials: L
authors.lastname: Chatel
authors.affiliation: Laboratory for Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry and Microbiology, Ghent University, Ghent, Belgium.
authors.forename: Bart
authors.initials: B
authors.lastname: Devreese
authors.affiliation: 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.
authors.forename: Nico
authors.initials: N
authors.lastname: Callewaert
mesh_terms: D000818:Animals; D006031:Glycosylation; D006801:Humans; D051379:Mice; D011134:Polysaccharides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D011134:Polysaccharides; D011994:Recombinant Proteins
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