Multimeric proteins and methods of making and using same
The invention provides multimerization polypeptides capable of conferring formation of multimers when the multimerization polypeptide is linked to a molecule, such as a heterologous polypeptide sequence.
1. A multimerization polypeptide comprising an amino acid sequence selected from the group consisting of (i) SEQ ID NO: 154; and (ii) SEQ ID NO: 154, wherein one or more of amino acid positions 1, 4, 8, 11, 15, 18, 22, 25, and 29 is substituted with leucine, isoleucine, or valine.
2. The multimerization polypeptide of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 154.
3. The multimerization polypeptide of claim 1 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 154, wherein one or more of amino acid positions 1, 4, 8, 11, 15, 18, 22, 25, and 29 is substituted with leucine, isoleucine, or valine.
4. The multimerization polypeptide of claim 2 , wherein the polypeptide consists of the amino acid sequence of SEQ ID NO: 154.
5. The multimerization polypeptide of claim 4 , wherein the polypeptide consists of the amino acid sequence of SEQ ID NO: 154, wherein one or more of amino acid positions 1, 4, 8, 11, 15, 18, 22, 25, and 29 is substituted with leucine, isoleucine, or valine.
6. A chimeric protein comprising a heterologous polypeptide fused to the N-terminus of the multimerization polypeptide of claim 1 .
7. The chimeric protein of claim 6 , wherein the heterologous polypeptide is selected from the group consisting of a binding protein, enzyme, receptor, ligand, nucleic acid binding protein, growth regulatory factor, differentiative factor, and chemotactic factor.
8. The chimeric protein of claim 7 , wherein the binding protein comprises an antigen binding polypeptide.
9. The chimeric protein of claim 8 , wherein the antigen binding polypeptide comprises at least one antibody variable domain.
10. The chimeric protein of claim 9 , wherein the antibody variable domain is human or humanized.
11. The chimeric protein of claim 8 , wherein the antigen binding polypeptide comprises a single chain antibody, Fab, Fab′, (Fab′) 2 , or Fv antibody subsequence.
12. The chimeric protein of claim 8 , wherein the antigen binding polypeptide comprises a multispecific or multifunctional antibody.
13. The chimeric protein of claim 8 , wherein the antigen binding polypeptide binds to ICAM-1 or an epitope thereof.
14. The chimeric protein of claim 6 , wherein said chimeric protein contains a linker between the heterologous polypeptide and multimerization polypeptide.
15. The chimeric protein of claim 14 , wherein the linker comprises a hinge domain.
16. The chimeric protein of claim 14 , wherein the linker comprises an amino acid sequence set forth in any of SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46 or SEQ ID NO: 47.
17. The chimeric protein of claim 14 , wherein the linker comprises an amino acid sequence having a length of from about 5 to 20 amino acids.
18. The chimeric protein of claim 14 , wherein the linker comprises an amino acid sequence having a length of from about 10 to 30 amino acids.
19. The chimeric protein of claim 14 , wherein the linker comprises an amino acid sequence having a length of from about 25 to 50 amino acids.
20. The chimeric protein of claim 14 , wherein the linker comprises an amino acid sequence having a length of from about 30 to 60 amino acids.
21. The chimeric protein of claim 14 , wherein the linker comprises an amino acid sequence having a length of from about 50 to 75 amino acids.
22. A polypeptide oligomer comprising at least two chimeric proteins of claim 6 .
23. The polypeptide oligomer of claim 22 that is a tetramer.
24. The polypeptide oligomer of claim 23 that is homomeric tetramer.
25. A pharmaceutical formulation comprising the chimeric protein of claim 6 and a pharmaceutical carrier.
26. The pharmaceutical formulation of claim 25 , wherein the pharmaceutical carrier is a sterile, aqueous solution.
27. The pharmaceutical formulation of claim 26 , wherein the pharmaceutical carrier is an ointment, salve, gel, or cream.
28. A kit comprising the pharmaceutical formulation of claim 26 in a container.
29. The kit of claim 28 , further comprising instructions for administration of the pharmaceutical formulation.