Dual variable domain immunnoglobulins and uses thereof

1. A binding protein comprising first and second polypeptide chains, 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

wherein the second polypeptide chain comprises a second VD1-(X1)n-VD2-C-(X2)n, 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;

X2 does not comprise an Fc region;

n is 0 or 1, and

wherein the binding protein is capable of binding a pair of antigens selected from the group consisting of IL-1alpha and IL-1beta,

wherein either the VD1 heavy and light chain variable domain or the VD2 heavy and light chain variable domain form an antigen binding domain that binds the IL-1beta antigen and comprises the amino acid sequences of SEQ ID NOs: 30 and 31 or SEQ ID NOs: 32 and 33, and

wherein the other of the VD1 or VD2 heavy and light chain variable domains forms an antigen binding domain that binds the IL-1alpha antigen.

2. The binding protein of claim 1 , wherein the heavy chain variable domain of the antigen binding domain that binds the IL-1alpha antigen comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 40, 42, 44, and 46, and wherein the light chain variable domain of the antigen binding domain that binds the IL-1alpha antigen comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 41, 43, 45, and 47.

3. The binding protein of claim 1 , wherein X1 or X2 is an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-29.

4. The binding protein of claim 1 , wherein the binding protein comprises two first polypeptide chains and two second polypeptide chains.

5. The binding protein of claim 1 , wherein the Fc region is a variant sequence Fc region.

6. A binding protein capable of binding two antigens comprising four polypeptide chains,

wherein two polypeptide chains comprise 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

wherein two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, 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;

X2 does not comprise an Fc region;

n is 0 or 1; and

wherein the VD1 heavy and light chain variable domains form an antigen binding domain that binds the IL-1beta antigen

and comprise the amino acid sequences of SEQ ID NOs 30 and 31 or SEQ ID NOs 32 and 33; and

wherein the VD2 heavy and light chain variable domains forms an antigen binding domain that binds the IL-1 alpha antigen.

7. A binding protein capable of binding two antigens comprising four polypeptide chains,

wherein two polypeptide chains comprise 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;

n is 0 or 1; and

wherein two polypeptide chains comprise VD1-(X1)n-VD2-C-(X2)n, 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;

X2 does not comprise an Fc region;

n is 0 or 1; and

wherein the VD2 heavy and light chain variable domains form an antigen binding domain that binds the IL-1beta antigen

and comprise the amino acid sequences of SEQ ID NOs 30 and 31 or SEQ ID NOs 32 and 33; and

wherein the VD1 heavy and light chain variable domains forms an antigen binding domain that binds the IL-1 alpha antigen.

8. The binding protein of any one of claims 1 , 6 , and 7 , wherein the binding protein has an on rate constant (Kon) to IL-1beta antigen selected from the group consisting of: at least about 10 2 M −1 s −1 ; at least about 10 3 M −1 s −1 ; at least about 10 4 M −1 s −1 ; at least about 10 5 M −1 s −1 ; and at least about 10 6 M −1 s −1 , as measured by surface plasmon resonance.

9. The binding protein of any one of claims 1 , 6 , and 7 , wherein the binding protein has an off rate constant (Koff) to IL-1beta antigen selected from the group consisting of: at most about 10 −3 s −1 ; at most about 10 −4 s −1 ; at most about 10 −5 s −1 ; and at most about 10 −6 s −1 , as measured by surface plasmon resonance.

10. The binding protein of any one of claims 1 , 6 , and 7 , wherein the binding protein has a dissociation constant (K D ) to IL-1beta antigen selected from the group consisting of: at most about 10 −7 M; at most about 10 −8 M; at most about 10 −9 M; at most about 10 −10 M; at most about 10 −11 M; at most about 10 −12 M; and at most 10 −13 M.

11. A binding protein conjugate comprising a binding protein of any one of claims 1 , 6 , and 7 , the binding protein conjugate further comprising an agent selected from the group consisting of an immunoadhesion molecule, an imaging agent, a therapeutic agent, and a cytotoxic agent.

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

13. The binding protein conjugate of claim 12 , wherein the imaging agent is a radiolabel selected from the group consisting of: 3 H, 14 C, 35 S, 90 Y, 99 Tc, 111 In, 125 I, 131 I, 177 Lu, 166 Ho, and 153 Sm.

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

15. A pharmaceutical composition comprising the binding protein of any one of claims 1 , 6 , and 7 , and a pharmaceutically acceptable carrier.

16. The pharmaceutical composition of claim 15 , wherein the composition further comprises at least one additional therapeutic agent.

17. The pharmaceutical composition of claim 16 , wherein the additional therapeutic agent is selected from the group consisting of: a cytotoxic agent, an angiogenesis inhibitor, a kinase inhibitor, a co-stimulation molecule blocker; an adhesion molecule blocker, an anti-cytokine antibody or functional fragment thereof, methotrexate, cyclosporin, rapamycin, FK506, a detectable label or reporter, a TNF antagonist, an antirheumatic, a muscle relaxant, a narcotic, a non-steroid anti-inflammatory drug (NSAID), an analgesic, an anesthetic, a sedative, a local anesthetic, a neuromuscular blocker, an antimicrobial, an antipsoriatic, a corticosteriod, an anabolic steroid, an erythropoietin, an immunization, an immunoglobulin, an immunosuppressive, a growth hormone, a hormone replacement drug, a radiopharmaceutical, an antidepressant, an antipsychotic, a stimulant, an asthma medication, a beta agonist, an inhaled steroid, an epinephrine or analog, a cytokine, and a cytokine antagonist.

18. The binding protein of claim 1 , wherein the VD1 or VD2 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 30 and 31.

19. The binding protein of claim 1 , wherein the VD1 or VD2 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 32 and 33.

20. The binding protein of claim 6 , wherein the VD1 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 30 and 31.

21. The binding protein of claim 6 , wherein the VD1 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 32 and 33.

22. The binding protein of claim 6 , wherein the VD2 heavy chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 40, 42, 44, and 46, and wherein the VD2 light chain variable domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 41, 43, 45, and 47.

23. The binding protein of claim 7 , wherein the VD2 heavy and light chain variable domains comprises the amino acid sequences of SEQ ID NOs 30 and 31.

24. The binding protein of claim 7 , wherein the VD2 heavy and light chain variable domains comprises the amino acid sequences of SEQ ID NOs 32 and 33.

25. The binding protein of claim 1 , wherein the VD1 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 30 and 31.

26. The binding protein of claim 1 , wherein the VD1 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 32 and 33.

27. The binding protein of claim 1 , wherein the VD2 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 30 and 31.

28. The binding protein of claim 1 , wherein the VD2 heavy and light chain variable domains comprise the amino acid sequences of SEQ ID NOs 32 and 33.