Compositions comprising polynucleotides encoding TACI-immunoglobulin fusion proteins and methods for producing the same
Molecules that interfere with the binding of a tumor necrosis factor receptor with its ligand, such as a soluble receptor, have proven usefulness in both basic research and as therapeutics. The present invention provides improved soluble transmembrane activator and calcium modulator and cyclophilin ligand-interactor (TACI) receptors.
1. A nucleic acid molecule construct encoding a fusion protein, comprising:
(a) a first polynucleotide encoding a transmembrane activator and calcium modulator and cyclophilin ligand-interactor (TACI) receptor moiety, wherein the TACI receptor moiety consists of amino acid residue 30 to 154 of SEQ ID NO:2 and wherein the TACI receptor moiety binds at least one of ZTNF2 or ZTNF4; and,
(b) a second polynucleotide encoding an immunoglobulin moiety that comprises a heavy chain constant region of an immunoglobulin;
wherein said first and said second polynucleotide encode said fusion protein.
2. The nucleic acid molecule construct of claim 1 , wherein said nucleic acid molecule construct is operably linked to a promoter that directs transcription of the encoded fusion protein.
3. The nucleic acid molecule construct of claim 1 , further comprising a third polynucleotide encoding a secretory sequence.
4. The nucleic acid molecule construct of claim 1 , wherein the immunoglobulin moiety comprises a human heavy chain constant region.
5. The nucleic acid molecule construct of claim 1 , wherein the heavy chain constant region is an IgGl heavy chain constant region.
6. The nucleic acid molecule construct of claim 1 , wherein the immunoglobulin moiety is an IgGl Fc fragment that comprises C H2 and C H3 domains.
7. The nucleic acid molecule construct of claim 1 , wherein the immunoglobulin moiety is an IgGl Fc fragment comprising the amino acid sequence of SEQ ID NO:33.
8. An expression vector comprising the nucleic acid construct of claim 1 .
9. A host cell comprising the nucleic acid construct of claim 1 .
10. A nucleic acid molecule construct encoding a fusion protein, comprising:
(a) a first polynucleotide encoding a transmembrane activator and calcium modulator and cyclophilin ligand-interactor (TACI) receptor moiety, wherein the TACI receptor moiety consists of amino acid residue 30 to 110 of SEQ ID NO:2 and wherein the TACI receptor moiety binds at least one of ZTNF2 or ZTNF4; and,
(b) a second polynucleotide encoding an immunoglobulin moiety that comprises a heavy chain constant region of an immunoglobulin;
wherein said first and said second polynucleotide encode said fusion protein.
11. The nucleic acid molecule construct of claim 10 , wherein said nucleic acid molecule construct is operably linked to a promoter that directs transcription of the encoded fusion protein.
12. The nucleic acid molecule construct of claim 10 , further comprising a third polynucleotide encoding a secretory sequence.
13. The nucleic acid molecule construct of claim 10 , wherein the immunoglobulin moiety comprises a human heavy chain constant region.
14. The nucleic acid molecule construct of claim 10 , wherein the heavy chain constant region is an IgGl heavy chain constant region.
15. The nucleic acid molecule construct of claim 10 , wherein the immunoglobulin moiety is an IgGl Fc fragment that comprises C H2 and C H3 domains.
16. The nucleic acid molecule construct of claim 10 , wherein the immunoglobulin moiety is an IgGl Fc fragment comprising the amino acid sequence of SEQ ID NO:33.
17. The nucleic acid molecule construct of claim 10 , wherein the encoded fusion protein has an amino acid sequence comprising the amino acid sequence of SEQ ID NO:54.
18. The nucleic acid construct of claim 10 , wherein said construct comprises the nucleotide sequence set forth in SEQ ID NO: 53.
19. The nucleic acid molecule construct of claim 10 , wherein the encoded fusion protein has an amino acid sequence comprising the secreted form of the amino acid sequence of SEQ ID NO:54.
20. An expression vector comprising the nucleic acid molecule construct of claim 10 .
21. An expression vector comprising the nucleic acid molecule construct of claim 17 .
22. A host cell comprising the nucleic acid molecule construct of claim 10 .
23. A host cell comprising the nucleic acid molecule construct of claim 17 .
24. A method of expressing a fusion protein comprising culturing a host cell comprising the nucleic acid molecule construct of claim 2 under conditions that allow for the expression of said fission protein.
25. The method of claim 24 farther comprising isolating said fission protein.
26. The nucleic acid molecule construct of claim 10 , wherein the TACT-immunoglobulin fusion protein comprises the amino acid sequence of SEQ ID NO: 54, wherein the optimized tPA (otPA) leader sequence (SEQ ID NO:25) has been removed.
27. The nucleic acid molecule construct of claim 17 , wherein said nucleic acid molecule construct is operably linked to a promoter that directs transcription of the encoded fusion protein.
28. The nucleic acid molecule construct of claim 18 , wherein said nucleic acid molecule construct is operably linked to a promoter that directs transcription of the encoded fusion protein.
29. The nucleic acid molecule construct of claim 19 , wherein said nucleic acid molecule construct is operably linked to a promoter that directs transcription of the encoded fusion protein.
30. The nucleic acid molecule construct of claim 26 , wherein said nucleic acid molecule construct is operably linked to a promoter that directs transcription of the encoded fusion protein.
31. An expression vector comprising the nucleic acid molecule construct of claim 18 .
32. An expression vector comprising the nucleic acid molecule construct of claim 19 .
33. An expression vector comprising the nucleic acid molecule construct of claim 26 .
34. A host cell comprising the nucleic acid molecule construct of claim 18 .
35. A host cell comprising the nucleic acid molecule construct of claim 19 .
36. A host cell comprising the nucleic acid molecule construct of claim 26 .
37. A method of expressing a fusion protein comprising culturing a host cell comprising the nucleic acid molecule construct of claim 11 under conditions that allow for the expression of said fusion protein.
38. The method of claim 37 , further comprising isolating said fusion protein.
39. A method of expressing a fusion protein comprising culturing a host cell comprising the nucleic acid molecule construct of claim 27 under conditions that allow for the expression of said fusion protein.
40. The method of claim 39 , further comprising isolating said fusion protein.
41. A method of expressing a fusion protein comprising culturing a host cell comprising the nucleic acid molecule construct of claim 28 under conditions that allow for the expression of said fusion protein.
42. The method of claim 41 , further comprising isolating said fusion protein.
43. A method of expressing a fusion protein comprising culturing a host cell comprising the nucleic acid molecule construct of claim 29 under conditions that allow for the expression of said fusion protein.
44. The method of claim 43 , further comprising isolating said fusion protein.
45. A method of expressing a fusion protein comprising culturing a host cell comprising the nucleic acid molecule construct of claim 30 under conditions that allow for the expression of said fusion protein.
46. The method of claim 45 , further comprising isolating said fusion protein.