Factor VIII glycoforms
AssigneeBayer HealthCare LLC
InventorsMyung-Sam Cho, Sham-Yuen Chan
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Abstract
The invention concerns glycosylated proteins having human factor VIII activity. In a preferred embodiment, the protein is glycosylated with oligosaccharides that include an alpha-(2,6)-linked sialic acid and a bisecting GlcNAc linked to a core beta-mannose.
Claims38
A method of producing an isolated glycosylated protein having factor VIII procoagulant activity comprising the amino acid sequence of SEQ ID NO:
- 1, the method comprising:
- obtaining cells comprising a heterologous nucleic acid encoding the protein;
- culturing the cells under plasma derived protein-free conditions to produce the protein, and
- isolating the protein thereby produced,
- wherein the protein comprises an N-linked oligosaccharide having an alpha-(2,6)-linked sialic acid.
A method of producing an isolated glycosylated protein having factor VIII procoagulant activity comprising the amino acid sequence of SEQ ID NO:
- 1, the method comprising:
- obtaining cells comprising a heterologous nucleic acid encoding the protein;
- culturing the cells under plasma derived protein-free conditions to produce the protein, and
- isolating the protein thereby produced,
- wherein the protein comprises an N-linked oligosaccharide comprising a bisecting GlcNAc linked to a core beta-mannose.
A method of producing an isolated glycosylated protein having factor VIII procoagulant activity comprising the amino acid sequence of SEQ ID NO:
- 1, the method comprising:
- culturing cells comprising a heterologous nucleic acid encoding the protein in plasma derived protein-free medium to express the protein, and
- isolating the protein thereby produced,
- wherein the protein is expressed at a level of about 2 μU/c/d or greater.
A method of producing an isolated glycosylated protein having factor VIII procoagulant activity comprising the amino acid sequence of SEQ ID NO:
- 1, the method comprising:
- culturing cells comprising a heterologous nucleic acid encoding the protein in serum free medium supplemented with human plasma proteins to express the protein, and
- isolating the protein thereby produced,
- wherein the protein is expressed at a level of about 10 μU/c/d or greater.
Dependencies are inferred from the claim text.