Fc VARIANTS
Disclosed are Fc-containing proteins comprising a binding region and a variant Fc region that can elicit one or more immune effector function and/or bind to an Fc receptor more effectively than a similar Fc-containing protein comprising a wild type Fc region. Also disclosed are nucleic acids encoding such Fc-containing proteins, methods for making such proteins, and methods of treatment utilizing such proteins.
1 - 52 . (canceled)
53 . An Fc-containing protein comprising a heterodimeric human IgG Fc region and a binding region, wherein the Fc region comprises an A chain and a B chain, which each comprise from 1 to 10 amino acid substitutions relative to a wild type human Fc polypeptide chain, wherein the Fc-containing protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V with a K D of less than or equal to one fifth of the K D with which a second protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V, wherein the second protein is the same as the Fc-containing protein except that it contains a wild type human IgG Fc region without substitutions.
54 . The Fc-containing protein of claim 53 , wherein the Fc-containing protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V with a K D of less than or equal to one tenth of the K D with which the second protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V.
55 . The Fc-containing protein of claim 53 , wherein the A chain and the B chain each comprise from 1 to 6 amino acid substitutions relative to a wild type human Fc polypeptide chain.
56 . The Fc-containing protein of claim 53 , which comprises charge pair substitutions.
57 . The Fc-containing protein of claim 56 , wherein the A chain comprises the substitutions K392D and K409D and the B chain comprises the substitutions E356K and D399K or vice versa.
58 . The Fc-containing protein of claim 53 , wherein the IgG Fc region is an IgG1 Fc region.
59 . The Fc-containing protein of claim 53 , wherein
(a) the A chain comprises Q311M and K334V substitutions and the B chain comprises L234Y, E294L, and Y296W substitutions or vice versa;
(b) the A chain comprises E233L, Q311M, and K334V substitutions and the B chain comprises L234Y, E294L, and Y296W substitutions;
(c) the A chain comprises L234I, Q311M, and K334V substitutions and the B chain comprises L234Y, E294L, and Y296W substitutions or vice versa;
(d) the A chain comprises S298T and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(e) the A chain comprises A330M and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(f) the A chain comprises A330F and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(g) the A chain comprises Q311M, A330M, and K334V substitutions and the B chain comprises L234Y, E294L, and Y296W substitutions or vice versa;
(h) the A chain comprises Q311M, A330F, and K334V substitutions and the B chain comprises L234Y, E294L, and Y296W substitutions or vice versa;
(i) the A chain comprises S298T, A330M, and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(j) the A chain comprises S298T, A330F, and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(k) the A chain comprises S239D, A330M, and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(l) the A chain comprises S239D, S298T, and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(m) the A chain comprises a K334V substitution and the B chain comprises Y296W and S298C substitutions or vice versa;
(n) the A chain comprises a K334V substitution and the B chain comprises L234Y, Y296W, and S298C substitutions or vice versa;
(o) the A chain comprises L235S, S239D, and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(p) the A chain comprises L235S, S239D, and K334V substitutions and the B chain comprises L234Y, Y296W, and S298C substitutions or vice versa;
(q) the A chain comprises Q311M and K334V substitutions and the B chain comprises L234Y, F243V, and Y296W substitutions or vice versa;
(r) the A chain comprises Q311M and K334V substitutions and the B chain comprises L234Y, K296W, and S298C substitutions or vice versa;
(s) the A chain comprises S239D and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(t) the A chain comprises S239D and K334V substitutions and the B chain comprises L234Y, Y296W, and S298C substitutions or vice versa;
(u) the A chain comprises F243V and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W, substitutions or vice versa;
(v) the A chain comprises F243V and K334V substitutions and the B chain comprises L234Y, Y296W, and S298C substitutions or vice versa;
(w) the A chain comprises E294L and K334V substitutions and the B chain comprises L234Y, K290Y, and Y296W substitutions or vice versa;
(x) the A chain comprises E294L and K334V substitutions and the B chain comprises L234Y, Y296W, and S298C substitutions or vice versa;
(y) the A chain comprises A330M and K334V substitutions and the B chain comprises L234Y and Y296W substitutions or vice versa; or
(z) the A chain comprises A330M and K334V substitutions and the B chain comprises K290Y and Y296W substitutions or vice versa.
60 . The Fc-containing protein of claim 59 , wherein the A chain comprises the amino acid sequence of SEQ ID NO: 8, 12, 14, 16, 20, 22, 24, 26, 28, 30, 32, 34, or 37 and the B chain comprises the amino acid sequence of SEQ ID NO: 10, 18, 39, or 41.
61 . The Fc-containing protein of claim 60 comprising a pair of amino acid sequences selected from the group consisting of: SEQ ID NO: 8 and SEQ ID NO:10; SEQ ID NO: 16 and SEQ ID NO:18; SEQ ID NO: 12 and SEQ ID NO:10; SEQ ID NO: 14 and SEQ ID NO:10; SEQ ID NO: 20 and SEQ ID NO:18; SEQ ID NO: 22 and SEQ ID NO:18; SEQ ID NO: 24 and SEQ ID NO:10; SEQ ID NO: 26 and SEQ ID NO:10; SEQ ID NO: 28 and SEQ ID NO:18; SEQ ID NO: 30 and SEQ ID NO:18; SEQ ID NO: 32 and SEQ ID NO:18; SEQ ID NO: 34 and SEQ ID NO:18; SEQ NO:37 and SEQ ID NO:39; and SEQ NO:37 and SEQ ID NO:41.
62 . The Fc-containing protein of claim 53 , wherein:
the Fc-containing protein is defucosylated, is made in a CHO cell, and/or is an antibody; or
the Fc-containing protein is defucosylated, is made in a CHO cell, and/or is an Fc fusion protein.
63 . The Fc-containing protein of claim 62 , wherein the Fc-containing protein is an antibody and wherein the antibody is bispecific and/or is a full length human IgG1 antibody.
64 . The Fc-containing protein of claim 53 , wherein the Fc-containing protein binds to HER-2/neu, wherein the Fc-containing protein is a bispecific antibody that binds to CD38 and CD138, or wherein the Fc-containing protein binds to mesothelin.
65 . A pharmaceutical composition comprising a therapeutically effective amount of the Fc-containing protein of claim 53 and a pharmaceutically acceptable carrier.
66 . One or more nucleic acids encoding the Fc-containing protein of claim 53 .
67 . The one or more nucleic acids of claim 66 , which comprise the nucleotide sequences of SEQ ID NO: 7, 11, 13, 15, 19, 21, 23, 25, 27, 29, 31, 33, or 36 and SEQ ID NO:9, 17, 38, or 40.
68 . A host cell comprising the one or more nucleic acids of claim 66 .
69 . The host cell of claim 68 , which is a CHO cell.
70 . A method of making an Fc-containing protein containing a heterodimeric Fc region comprising the following steps:
(a) providing a host cell containing one or more nucleic acids encoding an Fc-containing protein comprising a heterodimeric human IgG Fc region and a binding region, wherein the Fc region comprises an A chain and a B chain, which each comprise from 1 to 10 amino acid substitutions relative to a wild type human Fc polypeptide chain, wherein the Fc-containing protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V with a K D of less than or equal to one fifth of the K D with which a second protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V, wherein the second protein is the same as the Fc-containing protein except that it contains a wild type human IgG Fc region without substitutions;
(b) culturing the host cell under conditions such that the Fc-containing protein will be expressed; and
(c) recovering the Fc-containing protein from the cell mass or the culture medium.
71 . A method for treating cancer comprising administering an Fc-containing protein or pharmaceutical composition comprising the Fc-containing protein to a cancer patient, wherein the Fc-containing protein comprises a heterodimeric human IgG Fc region and a binding region, wherein the Fc region comprises an A chain and a B chain, which each comprise from 1 to 10 amino acid substitutions relative to a wild type human Fc polypeptide chain, wherein the Fc-containing protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V with a K D of less than or equal to one fifth of the K D with which a second protein binds to human FcγRIIIA-158F and/or FcγRIIIA-158V, wherein the second protein is the same as the Fc-containing protein except that it contains a wild type human IgG Fc region without substitutions.
72 . The method of claim 71 , wherein chemotherapy and/or radiation is administered before, after, or concurrently with the administration of the Fc-containing protein.