Methods of producing glycosylated proteins
AssigneesLimmatech Biologics AG, Eidgenossische Technische Hochschule Zurich
InventorsMichael L. Wetter, Michael T. Kowarik, Amirreza Faridmoayer, Manuela Mally, Christian A. Lizak, Markus Aebi, Chia-wei Lin, Ivan Hang, Timothy Keys
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Abstract
Described herein are methods of producing glycosylated proteins in vitro and in vivo. The methods include using host cells to produce glycosylated proteins. Also described herein are glycosylated proteins produced using such methods and uses thereof.
Claims13
A host cell comprising (i) a heterologous nucleic acid that encodes a target protein comprising an N-glycosylation consensus sequence of the formula Asn-X-Ser(Thr), wherein X can be any amino acid residue except Pro;
- (ii) a nucleic acid that encodes a heterologous N-glycosyltransferase (NGT) that adds glucose to an amino acid residue present in said N-glycosylation consensus sequence;
- (iii) a nucleic acid that encodes a heterologous galactosyltransferase that catalyzes addition of a galactose to said glucose;
- (iv) a nucleic acid that encodes a heterologous sialyltransferase;
- (v) a nucleic acid that encodes a heterologous polysialyltransferase (polyST);
- and (vi) a nucleic acid that encodes a heterologous CMP-Neu5Ac synthetase;
- wherein the host cell is capable of glycosylating the target protein at the N-glycosylation consensus sequence with glucose-galactose primer and polysialylation.
Dependencies are inferred from the claim text.