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US 8,795,984 B2Granted
Production of glycoproteins with reduced O-glycosylation
AssigneeMerck Sharp & Dohme Corp.
InventorsPiotr Bobrowicz, W. James Cook, Warren Kett
Life of this patent
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Abstract
A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more α-1,2-mannosidases.
Claims10
A method for producing a heterologous protein from a yeast host cell comprising:
- (a) providing a yeast host cell having a nucleic acid encoding the heterologous protein operably linked to an inducible promoter;
- (b) contacting the yeast host cell with an inducer of the promoter to induce expression of the heterologous protein at a time before or at the same time as contacting the yeast host cell with 5-[[3-(1-Phenyl-2-hydroxy)ethoxy)-4-(2-phenylethoxy)]phenyl]methylene]-4-oxo-2-thioxo-3-thiazolidineacetic Acid;
- and
- (e) isolating the heterologous protein produced by the host cell.
1 more element
A method for producing a heterologous protein from a yeast host cell comprising:
- (a) providing a yeast host cell having a nucleic acid encoding the heterologous protein operably linked to an inducible promoter and a second nucleic acid encoding a Trichoderma reesei α-1,2-mannosidase enzyme;
- (b) contacting the yeast host cell with an inducer of the promoter to induce expression of the heterologous protein at a time before or at the same time as contacting the yeast host cell with 5-[[3-(1-Phenyl-2-hydroxy)ethoxy)-4-(2-phenylethoxy)]phenyl]methylene]-4-oxo-2-thioxo-3-thiazolidineacetic Acid;
- and
- (e) isolating the heterologous protein produced by the host cell.
1 more element
Dependencies are inferred from the claim text.