Genetically engineered eukaryotic cells producing sialylated glycoproteins
AssigneesVib Vzw, Universiteit Gent
InventorsNico Callewaert, Wander Van Breedam, Francis Santens
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Abstract
The present application provides glycosylation engineered higher eukaryotic cells which produce homogeneous forms of sialylated glycans present on a recombinant glycoprotein. In particular, the glycosylation engineered higher eukaryotic cells produce homogeneous forms of small N-glycan structures which carry terminal sialic acid residues. In addition, the invention provides glycan-conjugates based on specific coupling with sialic acid residues present on the recombinant glycoproteins.
Claims17
A eukaryotic cell comprising:
- a first exogenous nucleic acid sequence encoding an endoglucosaminidase enzyme;
- a second nucleic acid sequence encoding a mutant UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase which is insensitive to CMP-Neu5Ac feedback inhibition;
- and
- a third exogenous nucleic acid sequence encoding a glycoprotein.
A composition comprising a plurality of glycosylated forms of a recombinant Fc-containing molecule, wherein glycosylated forms comprising N-glycans consisting of a GlcNAc residue, glycosylated forms comprising N-glycans consisting of LacNAc disaccharides and glycosylated forms comprising N-glycans consisting of sialyl-LacNAc trisaccharides of said Fc-containing molecule are present in the composition, and wherein the amount of glycosylated forms comprising N-glycans consisting of sialyl-LacNAc trisaccharides is at least 39% in the pool of glycosylated forms comprising N-glycans consisting of GlcNAc residues, glycosylated forms comprising N-glycans consisting of LacNAc disaccharides and glycosylated forms comprising N-glycans consisting of sialyl-LacNAc trisaccharides of the Fc-fusion protein.
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Dependencies are inferred from the claim text.