Dual variable domain immunoglobulins and uses thereof

1. A binding protein comprising first and second polypeptide chains, each independently comprising VD1-(X1)n-VD2-C-(X2)n, wherein

VD1 is a first variable domain;

VD2 is a second variable domain;

C is a constant domain;

X1 is a linker;

X2 is an Fc region; and

n is 0 or 1;

wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site, and wherein the binding protein is capable of binding NGF and PGE2, wherein

(1) the variable domains that form a functional target binding site for NGF comprise CDRs 1-3 from SEQ ID NO: 38 and CDRs 1-3 from SEQ ID NO: 39; and

(2) the variable domains that form a functional target binding site for PGE2 comprise CDRs 1-3 from SEQ ID NO: 32 and CDRs 1-3 from SEQ ID NO: 33.

2. A binding protein comprising first and second polypeptide chains, each independently comprising VD1-(X1)n-VD2-C-(X2)n, wherein

VD1 is a first variable domain;

VD2 is a second variable domain;

C is a constant domain;

X1 is a linker;

X2 is an Fc region; and

n is 0 or 1;

wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site, and wherein the binding protein is capable of binding NGF and PGE2, wherein

(1) the variable domains that form a functional target binding site for NGF comprise SEQ ID NO: 38 and/or SEQ ID NO: 39;

and

(2) the variable domains that form a functional target binding site for PGE2 comprise SEQ ID NO: 32 and/or SEQ ID NO: 33.

3. The binding protein according to claim 1 or 2 , wherein the first polypeptide chain comprises a first VD1-(X1)n-VD2-C-(X2)n, wherein

VD1 is a first heavy chain variable domain;

VD2 is a second heavy chain variable domain;

C is a heavy chain constant domain;

X1 is a linker;

X2 is an Fc region; and

n is 0 or 1;

wherein the second polypeptide chain comprises a second VD1-(X1)n-VD2-C, wherein

VD1 is a first light chain variable domain;

VD2 is a second light chain variable domain;

C is a light chain constant domain;

X1 is a linker; and

n is 0 or 1;

and wherein the VD1 domains on the first and second polypeptide chains form a first functional target binding site and the VD2 domains on the first and second polypeptide chains form a second functional target binding site.

4. The binding protein according to claim 1 , wherein

(1) the variable domains that form a functional target binding site for NGF comprise SEQ ID NO: 38 and SEQ ID NO: 39;

and

(2) the variable domains that form a functional target binding site for PGE2 comprise SEQ ID NO: 32 and SEQ ID NO: 33.

5. The binding protein according to claim 1 or 2 , comprising two first polypeptide chains and two second polypeptide chains, forming four functional target binding sites.

6. The binding protein according to claim 1 or 2 , wherein X1 is any one of SEQ ID NO: 1-26.

7. The binding protein according to claim 1 or 2 , wherein the Fc region is a variant sequence Fc region.

8. The binding protein according to claim 1 or 2 , wherein the Fc region is an Fc region from IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, or IgD.

9. The binding protein according to claim 1 or 2 , wherein the binding protein is a crystallized binding protein.

10. A binding protein capable of binding NGF and PGE2, wherein the binding protein comprises:

DVD933 (comprising SEQ ID NOs: 48 and 49),

DVD934 (comprising SEQ ID NOs: 50 and 51),

DVD935 (comprising SEQ ID NOs: 52 and 53),

DVD936 (comprising SEQ ID NOs: 54 and 55),

DVD937 (comprising SEQ ID NOs: 56 and 57),

DVD938 (comprising SEQ ID NOs: 58 and 59),

DVD939 (comprising SEQ ID NOs: 60 and 61), or

DVD940 (comprising SEQ ID NOs: 62 and 63).

11. A binding protein conjugate comprising a binding protein according claim 1 or 2 , said binding protein conjugate further comprising an immunoadhesion molecule, an imaging agent, a therapeutic agent, or a cytotoxic agent.

12. The binding protein conjugate of claim 11 , wherein the imaging agent is a radiolabel, an enzyme, a fluorescent label, a luminescent label, a bioluminescent label, a magnetic label, or biotin.

13. The binding protein conjugate of claim 12 , wherein said radiolabel is 3 H, 14 C, 35 S, 90 Y, 99 Tc, 111 In, 125 l, 131 l, 177 Lu, 166 Ho, or 153 Sm.

14. The binding protein conjugate of claim 11 , wherein said therapeutic or cytotoxic agent is an anti-metabolite, an alkylating agent, an antibiotic, a growth factor, a cytokine, an anti-angiogenic agent, an anti-mitotic agent, an anthracycline, a toxin, or an apoptotic agent.

15. An isolated nucleic acid encoding a binding protein amino acid sequence according to claim 1 .

16. A vector comprising the isolated nucleic acid according to claim 15 .

17. A host cell comprising the vector according to claim 16 , wherein the host cell is optionally selected from the group consisting of a prokaryotic cell, Escherichia coli , a eukaryotic cell, an animal cell, a plant cell, a fungal cell, a yeast cell, an Sf9 cell, a mammalian cell, an avian cell, an insect cell, a CHO cell, and a COS cell.

18. A method of producing a binding protein, comprising culturing the host cell of claim 17 in culture medium under conditions sufficient to produce the binding protein.

19. A pharmaceutical composition comprising the binding protein according to any one of claims 1 , 2 , and 10 , and a pharmaceutically acceptable carrier.

20. The pharmaceutical composition according to claim 19 , further comprising at least one additional therapeutic agent.

21. A method of determining the presence, amount, or concentration of NGF and/or PGE2 in a test sample by an immunoassay,

wherein the immunoassay comprises contacting the test sample with at least one binding protein and at least one detectable label; and

wherein the at least one binding protein comprises the binding protein of claim 1 .

22. A kit for assaying a test sample for the presence, amount, or concentration of NGF and/or PGE2 in the sample, said kit comprising:

(a) instructions for assaying the test sample for NGF and/or PGE2; and

(b) at least one binding protein comprising the binding protein of claim 1 or 2 .

23. The binding protein according to claim 1 or 2 , wherein the binding protein is capable of inhibiting NGF with an EC 50 , of at most about 49 pM, as measured in a neruoscreen assay, capable of binding to NGF with a K D of at most about 5.13×10 −12 M, as measured by surface plasmon resonance, and/or capable of inhibiting PGE2 with an EC 50 , of at most about 23 pM, as measured in an EP4 bioassay.