FC mutants
The present invention provides reagents, methods and systems for predicting the inhibitory activity of an antibody or variant thereof comprising: determining a binding affinity of the antibody or variant thereof to a Fc activating receptor; determining a binding affinity of the antibody or variant thereof to a Fc inhibitory receptor, and calculating the ratio of said activating binding affinity to said inhibitory binding affinity (A/I ratio), wherein the magnitude of said ratio is less than one (1).
1. An isolated polypeptide comprising the amino acid sequence of a modified SEQ ID NO: 1, wherein said modification comprises:
substitutions at positions 247, 252, 266, 278, 302, and 354 of SEQ ID NO: 1.
2. An isolated polypeptide comprising the amino acid sequence of a modified SEQ ID NO: 1, wherein the modification is:
P247L, M252L, V266L, Y278F, V302E, and S354T
wherein the amino acid sequence possesses at least 95% identity to a native sequence Fc region that has the sequence of amino acid resides 118-447 of SEQ ID NO: 1.
3. A purified antibody comprising a polypeptide having the amino acid sequence of the isolated peptide of claim 2 .
4. A purified antibody comprising a polypeptide having the amino acid sequence of the isolated peptide of claim 2 , wherein the polypetide
(1) has a higher affinity to FcγRIIB than a control antibody comprising SEQ ID NO: 1, or
(2) has a lower affinity to at least one of FcγRI, FcγRIIA 131H , FcγRIIA 131R , FcγIIIA FF, and FcγIIIA FV than the control antibody, or
(3) has lower A/I ratio or lower FcγRIIA 131H /FcγRIIB A/I ratio or lower FcγRIIA 131R /FcγRIIB A/I ratio than the control antibody.
5. The purified antibody of claim 4 , wherein the purified antibody comprises a light chain having the same sequence as a light chain in the control antibody.
6. The purified antibody of claim 3 , which is a monoclonal antibody, a humanized antibody, or a human antibody.
7. A method of inhibiting an immune response of a cell expressing an FcγRIIB receptor and at least one of FcγRI, FcγRIIA 131H , FcγRIIA 131R , FcγIIIA FF, and FcγIIIA FV receptors, the method comprising contacting the cell with said antibody of claim 3 , having a lower A/I ratio between at least one of FcγRI, FcγRIIA 131H , FcγRIIA 131R , FcγIIIA FF, and FcγIIIA FV receptors and FcγRIIB receptor.
8. The method of claim 7 , wherein said cell is an effector cell or a macrophage.
9. A purified antibody comprising a polypeptide having the amino acid sequence of the isolated peptide of claim 1 .
10. The purified antibody of claim 9 , which is a monoclonal antibody, a humanized antibody, or a human antibody.