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US 12,398,380 B2Granted
Methods of host cell modification
AssigneeGlaxosmithkline Biologicals SA
InventorsMichael Wacker, Michael Kowarik, Fabiana Fernandez
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Abstract
Described herein are methods of inserting nucleic acid sequences into host cells. Also described herein are genetically stable host cells comprising inserted nucleic acid sequences and methods of using such host cells in the generation of proteins.
Claims5
A method of producing an N-glycosylated protein, wherein the method comprises culturing an Escherichia coli (E. coli) host cell under conditions suitable for production of proteins, wherein the E. coli host cell genome comprises:
- (i) a heterologous DNA sequence that encodes an oligosaccharyl transferase;
- (ii) a heterologous DNA sequence of at least 12 kb that encodes a glycosyltransferase from Candida albicans wherein the glycosyltransferase from Candida albicans is capable of making a Candida albicans polysaccharide;
- and
- wherein the E. coli host cell further comprises a gene that encodes a carrier protein containing at least SEQ ID NO:
- 72 having the sequence D/E-X-N-Z-S/T, wherein X and Z are independently selected from any amino acid except proline.
2 more elements
Dependencies are inferred from the claim text.