1 . A heterodimeric antibody or fragment thereof comprising one or more substitutions in each of the following domains: a first CH 3 -domain, a second CH 3 -domain, a CH 1 -domain, a C L -domain, a V H -domain and a V L -domain, wherein the one or more substitutions introduce charged amino acids that are electrostatically unfavorable to homodimer formation and electrostatically favorable to heterodimer formation.
2 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the first CH 3 -domain or the second CH 3 -domain comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more positive-charged amino acids in a wild-type human IgG amino acid sequence are replaced with one or more negative-charged amino acids.
3 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the first CH 3 -domain or the second CH 3 -domain comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more negative-charged amino acids in a wild-type human IgG amino acid sequence are replaced with one or more positive-charged amino acids.
4 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the CH 1 -domain or the C L -domain comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more positive-charged amino acids in wild-type IgG amino acid sequence are replaced with one or more negative-charged amino acids.
5 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the CH 1 -domain or the C L -domain comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more negative-charged amino acids in wild-type IgG amino acid sequence are replaced with one or more positive-charged amino acids.
6 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the V H -domain or the V L -domain comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more positive-charged amino acids in wild-type IgG amino acid sequence are replaced with one or more negative-charged amino acids.
7 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the V H -domain or the V L -domain comprises an amino acid sequence differing from wild-type IgG amino acid sequence such that one or more negative-charged amino acids in wild-type IgG amino acid sequence are replaced with one or more positive-charged amino acids.
8 . The heterodimeric antibody or fragment thereof of any one of claims 1 - 7 , wherein the positive-charged amino acid is selected from the group consisting of lysine, histidine and arginine.
9 . The heterodimeric antibody or fragment thereof of any one of claims 1 - 7 , wherein the negative-charged amino acid is selected from the group consisting of aspartic acid and glutamic acid.
10 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the first CH 3 -domain comprises a replacement of a glutamic acid with a positive-charged amino acid.
11 . The heterdimeric antibody or fragment thereof of claim 10 , wherein the glutamic acid is Glu356.
12 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the first CH 3 -domain comprises a replacement of an aspartic acid with a negative-charged amino acid.
13 . The heterodimeric antibody or fragment thereof of claim 12 , wherein the aspartic acid is Asp356 and Asp399.
14 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the second CH 3 -domain comprises a replacement of a lysine with a negative-charged amino acid.
15 . The heterodimeric antibody or fragment thereof of claim 14 , wherein the lysine is selected from the group consisting of Lys392, Lys409, Lys439 and Lys370.
16 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the CH 1 -domain comprises a replacement of a serine with a negative-charged amino acid.
17 . The heterodimeric antibody or fragment thereof of claim 16 , wherein the serine is Ser183.
18 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the C L -domain comprises a replacement of a serine with a positive-charged amino acid.
19 . The heterodimeric antibody or fragment thereof of claim 18 , wherein the serine is Ser176.
20 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the V H -domain comprises a replacement of a glutamine with a negative-charged amino acid.
21 . The heterodimeric antibody or fragment thereof of claim 20 , wherein the glutamine is Gln38 (Kabat position 38, AHo position 46).
22 . The heterodimeric antibody or fragment thereof of claim 1 , wherein the V L -domain comprises a replacement of a glutamine with a positive-charged amino acid.
23 . The heterodimeric antibody or fragment thereof of claim 22 , wherein the glutamine is Gln39 (Kabat position 39, AHo position 46).
24 . The heterodimeric antibody or fragment thereof of any one of claims 1 - 23 , that is selected from the group consisting of an antibody, a bispecific antibody, a monospecific monovalent antibody, a bispecific maxibody, a monobody, a peptibody, a bispecific peptibody, a monovalent peptibody and a receptor fusion protein.
25 . A heterodimeric antibody or fragment thereof comprising a heavy chain comprising (a) a first amino acid substitution at an AHo position selected from the group consisting of AHo positions 42-50 that introduces a charged amino acid at said position, (b) a second amino acid substitution at a position selected from the group consisting of positions 126-213 (EU numbering) that introduces a charged amino acid at said position, (c) a third amino acid substitution at a position selected from the group consisting of positions 352-360 (EU numbering) that introduces a charged amino acid at said position, and (d) a fourth amino acid substitution at a position selected from the group consisting of positions 395-403 (EU numbering) that introduces a charged amino acid, wherein the charged amino acid of (a) has the same charge as the charged amino acid of (b), and wherein the charged amino acids of (c) and (d) have an opposite charge of the charged amino acids of (a) and (b).
26 . The heterodimeric antibody or fragment thereof of claim 25 , wherein the first amino acid substitution is at AHo position 46, the second amino acid substitution is at EU position 183, the third amino acid substitution is at EU position 356 and the fourth amino acid substitution is at EU position 399.
27 . The heterodimeric antibody or fragment of claim 25 , wherein glutamine at AHo position 46 is replaced with glutamic acid, serine at EU position 183 is replaced with glutamic acid, glutamic acid at EU position 356 is replaced with lysine, and aspartic acid at EU position 399 is replaced with lysine.
28 . An antibody or fragment thereof comprising a heavy chain comprising (a) a first amino acid substitution at an AHo position selected from the group consisting of AHo positions 35-43 that introduces a charged amino acid at said position, (b) a second amino acid substitution at a position selected from the group consisting of positions 126-213 (EU numbering) that introduces a charged amino acid at said position, (c) a third amino acid substitution at a position selected from the group consisting of positions 388-398 (EU numbering) that introduces a charged amino acid at said position, and (d) a fourth amino acid substitution at a position selected from the group consisting of positions 404-413 (EU numbering) that introduces a charged amino acid, wherein the charged amino acids of (c) and (d) have an opposite charge of the charged amino acids of (a) and (b).
29 . The heterodimeric antibody or fragment thereof according to claim 28 , wherein the first amino acid substitution is at AHo position 46, the second amino acid substitution is at EU position 183, the third amino substitution is at EU position 393 and the fourth amino acid substitution is at EUposition 409.
30 . The antibody or fragment thereof claim 29 , wherein glutamine at AHo position 46 is replaced with lysine, serine at EUposition 183 is replaced with lysine, lysine at EU position 392 is replaced with aspartic acid and lysine at EU position 409 is replaced with aspartic acid.
31 . An heterodimeric antibody or fragment comprising a light chain comprising (a) a first amino acid substitution at an AHo position selected from the group consisting of AHo positions 42-50 that introduces a charged amino acid at said position, and (b) a second amino acid substitution at a position selected from the group consisting of positions 126-213 (EU numbering) that introduces a charged amino acid at said position.
32 . The heterodimeric antibody or fragment thereof of claim 31 , wherein the first amino acid substitution is at AHo position 46 and the second amino acid substitution is at EU position 176.
33 . The heterodimeric antibody or fragment of claim 32 , wherein glutamine at AHo position 46 is replaced with glutamic acid and serine at EU position 176 is replaced with glutamic acid.
34 . An antibody or fragment thereof comprising a heavy chain comprising (a) a first amino acid substitution at an AHo position selected from the group consisting of AHo positions 42-50 that introduces a charged amino acid at said position, (b) a second amino acid substitution at a position selected from the group consisting of positions 126-213 (EU numbering) that introduces a charged amino acid at said position, (c) a third amino acid substitution at a position selected from the group consisting of positions 352-360 (EU numbering) that introduces a charged amino acid at said position, and (d) a fourth amino acid substitution at a position selected from the group consisting of positions 395-403 (EU numbering) that introduces a charged amino acid, wherein the charged amino acids of (a) and (b) have a negative-charge and the charged amino acids of (c) and (d) have a positive charge.
35 . The antibody of claim 34 , wherein the first amino acid substitution is at position AHo 46, the second amino acid substitution is at EU position 183, the third amino acid substitution is at EU position 356 and the fourth amino acid substitution is at EU position 399.
36 . The antibody or fragment thereof of claim 34 , comprising a light chain comprising positive-charged amino acids at AHo position 46 and EU position 176.
37 . A heterodimeric antibody or fragment thereof comprising a heavy chain comprising (a) a first amino acid substitution at an AHo position selected from the group consisting of AHo positions 42-50 that introduces a charged amino acid at said position, (b) a second amino acid substitution at a position selected from the group consisting of positions 126-213 (EU numbering) that introduces a charged amino acid at said position, (c) a third amino acid substitution at a position selected from the group consisting of positions 388-397 (EU numbering) that introduces a charged amino acid at said position, and (d) a fourth amino acid substitution at a position selected from the group consisting of positions 404-413 (EU numbering) that introduces a charged amino acid, wherein the charged amino acids of (a) and (b) have a positive-charge and the charged amino acids of (c) and (d) have a negative-charge.
38 . The heterodimeric antibody or fragment thereof of claim 38 , wherein the first amino acid substitution is at AHo position 46, the second amino acid substitution is at EU position 183, the third amino acid substitution is at EU position 392 and the fourth amino acid substitution is at EU position 409.
39 . The heterodimeric antibody or fragment thereof of claim 37 , comprising a light chain comprising negative-charged amino acids at AHo position 46 and EU position 176.
40 . A heterodimeric antibody comprising a first heavy chain and a second heavy chain and a first light chain and a second light chain, wherein the first heavy chain comprises amino acid substitutions at AHo position 46 and EU positions 183, 356 and 399, wherein the second heavy chain comprises amino acid substitutions at AHo position 46 and EU positions 183, 392 and 409, and wherein the first and second light chains comprise an amino acid substitution at AHo position 46 and EU position 176, wherein the amino acid substitutions introduce a charged amino acid at said positions.
41 . The heterodimeric antibody of claim 40 , wherein glutamine at position AHo 46 of the first heavy chain is replaced with glutamic acid, wherein glutamine at position AHo 46 of the second heavy chain is replaced with lysine, wherein glutamine at position AHo 46 of the first light chain is replaced with lysine, wherein glutamine at position AHo 46 of the second light chain is replaced with glutamic acid, wherein serine at EU position 183 of the first heavy chain is replaced with glutamic acid, wherein glutamic acid at EU position 356 of the first heavy chain is replaced with lysine, wherein glutamic acid at EU position 399 of the first heavy chain is replaced with lysine, wherein serine at EU position 183 of the second heavy chain is replaced with lysine, wherein lysine at EU position 392 of the second heavy chain is replaced with aspartic acid, and wherein lysine at EU position 409 of the second heavy chain is replaced with aspartic acid.
42 . A heterodimeric antibody that binds sclerostin and DKK-1, comprising a first heavy chain comprising a variable region amino acid sequence selected from the group consisting of SEQ ID NOs: 378 and 366 and comprising amino acid substitutions at EU positions 183, 356 and 399 of the first heavy chain, a second heavy chain comprising a variable region amino acid sequence selected from the group consisting of SEQ ID NOs: 1003 and 974 and comprising amino acid substitutions at EU positions 183, 392 and 409 of the second heavy chain, a first light chain comprising a variable region amino acid sequence selected from the group consisting of SEQ ID NOs: 376 and 364 and comprising an amino acid substitution at EU position 176 of the first light chain, and a second light chain comprising a variable region amino acid sequence selected from the group consisting of SEQ ID NOs: 1002 and 978 and comprising an amino acid substitution at EU position 176 of the second light chain; wherein the amino acid substitutions introduce a charged amino acid at said positions.
43 . An antibody that binds to a region of sclerostin comprising amino acids 86-111 of SEQ ID NO: 1, wherein the antibody comprises substitutions in each of the following domains: a first CH 3 -domain, a second CH 3 -domain, a CH 1 -domain, and a C L -domain, wherein the one or more substitutions introduce charged amino acids that are electrostatically unfavorable to homodimer formation and electrostatically favorable to heterodimer formation.
44 . An antibody that binds to a region of sclerostin comprising amino acids 86-111 of SEQ ID NO: 1, wherein the antibody comprises a heavy chain having a CH3 domain comprising one or more amino acid substitutions, wherein the one or more substitutions introduce charged amino acids that are electrostatically unfavorable to homodimer formation and electrostatically favorable to heterodimer formation.
45 . The antibody of claim 44 , wherein a negative charged amino acid in the CH3 domain is substituted with a positive charged amino acid.
46 . The antibody of claim 45 , wherein the negative charged amino acid is at EU position D399, E356, or E357.
47 . The antibody of claim 45 , wherein the negative charged amino acid at EU positions D399, E356 and E357 are each substituted with a positive charged amino acid.
48 . The antibody of claim 45 or claim 47 , wherein the positive charged amino acid is lysine.
49 . The antibody of claim 44 , wherein a positive charge amino acid in the CH3 domain is substituted with a negative charged amino acid.
50 . The antibody of claim 49 , wherein the positive charged amino acid is at EU position K370, K392 or K409.
51 . The antibody of claim 49 , wherein the positive charged amino acids at EU positions K370, K392 and K409 are each substituted with a negative charged amino acid.
52 . The antibody of claim 49 or claim 51 , wherein the negative charged amino acid is aspartic acid.
53 . A heterodimeric antibody that binds to a region of sclerostin comprising amino acids 86-111 of SEQ ID NO: 1, wherein the antibody comprises a first heavy chain and a first light chain comprising a sclerostin-binding portion and a second heavy chain and a second light chain comprising a DKK1-binding portion, wherein the first heavy chain comprises amino acid substitutions at AHo position 46 and EU positions 183, 356 and 399, wherein the second heavy chain comprises amino acid substitutions at AHo position 46 and EU positions 183, 392 and 409, wherein the first and second light chains comprise an amino acid substitution at AHo position 46 and EU position 176, and wherein the amino acid substitutions introduce a charged amino acid at said positions.
54 . A heterodimeric antibody that binds to a region of sclerostin comprising amino acids 86-111 of SEQ ID NO: 1, the antibody comprising a first heavy chain and a first light chain comprising a sclerostin-binding portion and a second heavy chain and a second light chain comprising a DKK1-binding portion, wherein the first heavy chain comprises amino acid substitutions at EU positions 183, 356 and 399, wherein the second heavy chain comprises amino acid substitutions at EU positions 183, 392 and 409, and wherein the first and second light chains comprise an amino acid substitution at EU position 176, wherein the amino acid substitutions introduce a charged amino acid at said positions.
55 . The heterodimeric antibody of any one of claims 1 - 43 , wherein the antibody binds to a region of sclerostin comprising amino acids 86-111 of SEQ ID NO: 1.
56 . The heterodimeric antibody of any one of claims 1 - 55 , that is an IgG immunoglobulin.
57 . A nucleic acid comprising a nucleotide sequence encoding the heterodimeric antibody according to any one of claims 1 - 56 .
58 . A vector comprising the nucleotide sequence of claim 57 .
59 . An isolated host cell comprising the nucleic acid of claim 57 or the vector of claim 58 .
60 . A method of increasing bone mineral density in a mammalian subject comprising administering the heterodimeric antibody of any one of claims 1 - 56 to the subject in an amount effective to increase bone mineral density in the subject.
61 . A composition comprising the antibody of any one of claims 1 - 56 and a pharmaceutically acceptable carrier, diluent or adjuvant.
62 . The composition according to claim 61 , wherein less than 5% of the antibody in the composition is in aggregate form after two weeks of storage at about 4° C.
63 . The composition according to claim 61 , wherein less than 1% of the antibody in the composition is in aggregate form after two weeks of storage at about 4° C.
64 . The composition according to claim 62 or claim 63 , wherein an amount of antibody in the composition present in aggregate form in determined by Size Exclusion Chromatography.
65 . A composition comprising a heterodimeric antibody or fragment thereof and a pharmaceutically acceptable carrier, diluent or adjuvant, the heterodimeric antibody or fragment thereof comprising one or more substitutions in each of the following domains: a first CH 3 -domain, a second CH 3 -domain, a CH 1 -domain, a C L -domain, a V H -domain and a V L -domain, wherein the one or more substitutions introduce charged amino acids that are electrostatically unfavorable to homodimer formation and electrostatically favorable to heterodimer formation; wherein less than 5% of the antibody or fragment in the composition is in aggregate form after two weeks of storage at about 4° C.
66 . A composition comprising a heterodimeric antibody that binds to a region of sclerostin comprising amino acids 86-111 of SEQ ID NO: 1 and a pharmaceutically acceptable carrier, diluent or adjuvant, the antibody comprising substitutions in each of the following domains: a first CH 3 -domain, a second CH 3 -domain, a CH 1 -domain, and a C L -domain, wherein the one or more substitutions introduce charged amino acids that are electrostatically unfavorable to homodimer formation and electrostatically favorable to heterodimer formation; wherein less than 5% of the antibody in the composition is in aggregate form after two weeks of storage at about 4° C.
67 . The composition according to claim 66 or claim 67 , wherein less than 1% of the antibody in the composition is in aggregate form after two weeks of storage at about 4° C.
