Fucosylation-Deficient Cells
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsGang CHEN, Darya Burakov, James P. Fandl
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Abstract
An isolated nucleic acid encoding an FX protein having a serine at position 79, a lysine at position 90, a leucine at position 136, an arginine at position 211, a serine at position 289, and a combination thereof is provided. Cells having a gene encoding a modified FX protein are provided, wherein the cells exhibit a reduced ability to fucosylate a glycoprotein at a first temperature, but exhibit the ability to fucosylate the glycoprotein at a second temperature. Methods and compositions for making glycoproteins with reduced fucosylation are provided.
Claims20
A cell capable of fucosylating a glycoprotein, wherein the cell comprises a modified GDP-4-keto-6-deoxy-mannose-3,5-epimerase-4-reductase (FX) gene having a modification that codes for an FX protein that is at least 90% identical to SEQ ID NO:1 and that comprises a serine at position 289, and wherein no more than about 10% of the glycoprotein is fucosylated when the cell is grown at a temperature of about 37° C. in the absence of an external fucose source.
An isolated nucleotide sequence encoding an FX protein, wherein the FX protein comprises a serine at position 289.
A method for making a glycoprotein with reduced fucosylation, comprising:
- culturing a mammalian cell that comprises a FX gene that codes for a FX protein with a serine at position 289, at a temperature of about 37° C. and in the absence of an external fucose source, wherein the cell expresses a glycoprotein that comprises an immunoglobulin CH2 region and an immunoglobulin CH3 region;
- and, isolating the glycoprotein from the culture.
Dependencies are inferred from the claim text.