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AssigneeGlycoFi, Inc.
InventorsPiotr Bobrowicz, Stephen R. Hamilton, Tillman U. Gerngross, Stefan Wildt, Byung-Kwon Choi, Juergen H. Nett, Robert C. Davidson
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The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells exhibit GnTIII activity, which produce bisected N-glycan structures and may be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar transporters and mannosidases, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained.
A yeast host cell that has been genetically engineered to be diminished or depleted in the activity of an initiating α-1,6-mannosylatransferase and to comprise a nucleic acid sequence encoding an α-1,2-mannosidase, a nucleic acid sequence encoding a N-acetylglucosaminyltransferase I (GnT I), and a nucleic acid sequence encoding an N-acetylglucosaminyltransferase III (GnT III).
A yeast host cell that has been genetically engineered to be diminished or depleted in the activity of an initiating α-1,6-mannosylatransferase and to comprise a nucleic acid sequence encoding an α-1,2-mannosidase, a nucleic acid sequence encoding an N-acetylglucosaminyltransferase I (GnT I), a nucleic acid sequence encoding an N-acetylglucosaminyltransferase II (GnT II), and a nucleic acid sequence encoding an N-acetylglucosaminyltransferase III (GnT III).
A yeast host cell that has been genetically engineered to be diminished or depleted in the activity of an initiating α-1,6-mannosylatransferase and to comprise a nucleic acid sequence encoding an α-1,2-mannosidase, a nucleic acid sequence encoding a N-acetylglucosaminyltransferase I (GnT I), a nucleic acid sequence encoding an N-acetylglucosaminyltransferase III (GnT III), and a nucleic acid sequence encoding a mannosidase II.
Dependencies are inferred from the claim text.