Materials and methods for improved immunoglycoproteins
Immunoglycoproteins, including antibodies, with improved ADCC and altered glycosylation patterns are provided. Also provided are cell culturing methods and media for producing such immunoglycoproteins, and therapeutic uses of such immunoglycoproteins.
1. A composition comprising recombinant immunoglycoprotein molecules, wherein the recombinant immunoglycoprotein molecules each comprise an N-linked glycosylation site for linkage of oligosaccharide, and wherein at least 60% of the N-linked oligosaccharides of the N-linked glycosylation site have a hexose content of at least ten hexose molecules.
2. The composition of claim 1 , wherein the immunoglycoprotein molecules exhibit at least 5-fold higher ADCC compared to control immunoglycoprotein molecules of the same encoded amino acid sequence produced in CHO-K1 cells in the absence of a carbohydrate modifier.
3. The composition of claim 1 , wherein at least 60% of the N-linked oligosaccharides of the N-linked glycosylation site contain no fucose.
4. The composition of claim 1 formulated in a pharmaceutically acceptable carrier or diluent.
5. The composition of claim 4 , wherein the composition is sterile.
6. The composition of claim 1 comprising at least 100 g of immunoglycoprotein molecules and a pharmaceutically acceptable carrier or diluent.
7. The composition of claim 1 comprising at least 100 liters of culture medium.
8. The composition of any one of claims 1 - 3 , wherein the recombinant immunoglycoprotein molecules are bispecific in antigen-binding.
9. The composition of any one of claims 1 - 3 , wherein the recombinant immunoglycoprotein molecules are trispecific in antigen-binding.
10. The composition of any one of claims 1 - 3 , wherein the recombinant immunoglycoprotein molecule is an antibody or a small modular immunopharmaceutical.
11. The composition of claim 10 , wherein the small modular immunopharmaceutical comprises amino acids 24-496 of SEQ ID NO:2.