Method for preparing antibodies having improved properties
View Patent ↗The present invention is directed to methods and compositions for the production of Fc-containing polypeptides having improved properties and comprising mutations at positions 243 and 264 of the Fc region.
1. An Fc-containing polypeptide comprising the amino acid sequence of SEQ ID NO: 18, wherein said SEQ ID NO:18 comprises mutations at amino acid positions 243 and 264 of the Fc region, wherein the mutations are F243A and V264A, wherein the numbering is according to the EU index as in Kabat, and wherein the Fc-containing polypeptide is an antibody or antibody fragment comprising sialylated N-glycans.
2. The Fc-containing polypeptide of claim 1 , wherein the sialic acid residues in the sialylated N-glycans are attached via α-2,6 linkages.
3. The Fc-containing polypeptide of claim 1 or 2 , wherein the Fc-containing polypeptide has one or more of the following properties when compared to a parent Fc-containing polypeptide:
a) reduced effector function,
b) increased anti-inflammatory properties,
c) increased sialylation,
d) increased bioavailability when administered parenterally, and
e) reduced binding to FcγRI, FcγRIIa, FcγRIIb and FcγRIIIa.
4. The Fc-containing polypeptide of claim 1 , wherein at least 70 mole % of the N-glycans in the Fc-containing polypeptide have a structure selected from the group consisting of SA(1-4)Gal(1-4)GlcNAc((2-4)Man3GlcNAc2.
5. The Fc-containing polypeptide of claim 1 , wherein at least 90 mole % of the N-glycans in the Fc-containing polypeptide have a structure selected from the group consisting of SA(1-4)Gal(1-4)GlcNAc((2-4)Man3GlcNAc2.
6. The Fc-containing polypeptide of claim 1 , wherein at least 66 mole % of the N-glycans in the Fc-containing polypeptide have a NANA2Gal2GlcNAc2Man3GlcNAc2 structure.
7. The Fc-containing polypeptide of claim 1 , wherein at least 90 mole % of the N-glycans in the Fc-containing polypeptide have a structure selected from the group consisting of SA(1-4)Gal(1-4)GlcNAc((2-4)Man3GlcNAc2.
8. The Fc-containing polypeptide of claim 1 , wherein at least 66 mole % of the N-glycans in the Fc-containing polypeptide have a NANA2Gal2GlcNAc2Man3GlcNAc2 structure.