Fc containing polypeptides with altered glycosylation and reduced effector function
Provided are binding polypeptides (e.g., antibodies), and drug conjugates thereof, comprising an Fc domain with an altered glycosylation profile and reduced effector function. In particular embodiment, the Fc domain comprises: an asparagine residue at amino acid position 298, according to EU numbering; and a serine or threonine residue at amino acid position 300, according to EU numbering. Also provided are nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided.
1. An isolated polynucleotide encoding a binding polypeptide comprising an Fc domain, wherein the Fc domain comprises: an asparagine residue at amino acid position 298, according to EU numbering; and a serine or threonine residue at amino acid position 300, according to EU numbering, and wherein the encoded binding polypeptide has a lower affinity for an Fcγ receptor than a binding polypeptide having a native Fc domain.
2. A vector comprising the polynucleotide of claim 1 .
3. A host cell comprising the polynucleotide or vector of claim 1 .
4. A method of making a binding polypeptide comprising expressing the polynucleotide of claim 1 .
5. A method of making a binding polypeptide comprising expressing the vector of claim 2 in a cell.
6. The isolated polynucleotide of claim 1 , wherein the encoded binding polypeptide further comprises an alanine residue at amino acid position 299, according to EU numbering.
7. The isolated polynucleotide of claim 1 , wherein the encoded binding polypeptide further comprises a glutamine residue at amino acid position 297, according to EU numbering.
8. The isolated polynucleotide of claim 1 , wherein the Fc domain of the encoded polypeptide is an IgG1 Fc domain.
9. The isolated polynucleotide of claim 1 , wherein the Fc domain of the encoded polypeptide is human.
10. The isolated polynucleotide of claim 1 , wherein the Fcγ receptor is FcγRI and/or FcγRIIIa.
11. The isolated polynucleotide of claim 1 , wherein the encoded polypeptide has a similar affinity for an FcRn receptor as a binding polypeptide having a native Fc domain.
12. The isolated polynucleotide of claim 1 , wherein the encoded polypeptide is an antibody.
13. An isolated polynucleotide encoding a binding polypeptide comprising an Fc domain, wherein the Fc domain comprises: a free asparagine residue at amino acid position 298, according to EU numbering; and a free serine or threonine residue at amino acid position 300, according to EU numbering.
14. An isolated polynucleotide encoding a binding polypeptide comprising an Fc domain, wherein the Fc domain comprises: an N-glycan linked to an asparagine residue at amino acid position 298, according to EU numbering; and a free serine or threonine residue at amino acid position 300, according to EU numbering.
15. The isolated polynucleotide of claim 1 , wherein the encoded polypeptide is an immunoadhesin comprising a non-antibody binding region.
16. The isolated polynucleotide of claim 15 , wherein the non-antibody binding region of the encoded polypeptide is a receptor.
17. The isolated polynucleotide of claim 15 , wherein the non-antibody binding region of the encoded polypeptide is a ligand of a receptor.
18. The isolated polynucleotide of claim 1 , wherein the encoded polypeptide comprises at least one binding site which is a receptor binding site of a ligand.
19. An isolated polynucleotide encoding a binding polypeptide comprising a human IgG1 Fc domain, wherein the Fc domain comprises: an asparagine residue at amino acid position 297, according to EU numbering; an asparagine residue at amino acid position 298, according to EU numbering; an amino acid at position 299, according to EU numbering, wherein said amino acid at position 299 is not proline; and a serine or threonine residue at amino acid position 300, according to EU numbering.
20. An isolated polynucleotide encoding a binding polypeptide comprising a human IgG1 Fc domain, wherein the Fc domain comprises: a glutamine residue at amino acid position 297, according to EU numbering; an asparagine residue at amino acid position 298, according to EU numbering; an amino acid at position 299, according to EU numbering, wherein said amino acid at position 299 is not proline; and a serine or threonine residue at amino acid position 300, according to EU numbering.
21. An isolated polynucleotide encoding a binding polypeptide comprising a human IgG1 Fc domain, wherein the Fc domain comprises: an asparagine residue at amino acid position 297, according to EU numbering; an asparagine residue at amino acid position 298, according to EU numbering; an alanine residue at amino acid position 299, according to EU numbering; and a serine or threonine residue at amino acid position 300, according to EU numbering.
22. An isolated polynucleotide encoding a binding polypeptide comprising a human IgG4 Fc domain, wherein the Fc domain comprises: an asparagine residue at amino acid position 298, according to EU numbering; an alanine residue at amino acid position 299, according to EU numbering; and a serine or threonine residue at amino acid position 300, according to EU numbering.
23. The isolated polynucleotide of claim 22 , wherein the Fc domain of the encoded polypeptide comprises a hinge region with a Ser228Pro mutation (EU numbering).