Methods for chemically synthesizing immunoglobulin chimeric proteins
View Patent ↗The invention provides methods of chemically synthesizing chimeric proteins comprising at least a portion of an immunoglobulin constant region and a biologically active molecule.
1. A method of producing a chimeric protein comprising combining
(a) at least a portion of an immunoglobulin constant region comprising an FcRn binding site and having a naturally occurring cysteine from the interchain region of the immunoglobulin as the N-terminal amino acid; and
(b) a biologically active molecule comprising a functional group capable of reacting with an N terminal cysteine;
wherein the combining of (a) and (b) produces the chimeric protein.
2. The method of claim 1 , wherein the chimeric protein is chosen from a dimer and a monomer-dimer hybrid.
3. The method of claim 2 , wherein the monomer-dimer hybrid is a dimerically linked monomer-dimer hybrid.
4. The method of claim 1 , wherein the biologically active molecule is a polypeptide.
5. The method of claim 1 , wherein the functional group is thioester.
6. The methods of claim 5 , wherein the biologically active molecule is a polypeptide comprising an C terminal thioester.
7. The method of claim 1 , wherein the functional group is an aldehyde.
8. The method of claim 1 , wherein both the biologically active molecule and the portion of an immunoglobulin are produced recombinantly.
9. The method of claim 1 , wherein at least one of the biologically active molecule and the portion of an immunoglobulin is produced by chemical synthesis.
10. The method of claim 1 , wherein the chimeric protein further comprises a linker.
11. The method of claim 1 , wherein the reaction is performed in solution.
12. The method of claim 1 , wherein the reaction is performed on a solid support.
13. The method of claim 1 , wherein the biologically active molecule is chosen from EPO, interferon, a viral fusion inhibitor, and a clotting factor.
14. A method of producing a monomer-dimer hybrid protein comprising combining
(a) at least a portion of an immunoglobulin constant region comprising an FcRn binding site and having a naturally occurring cysteine from the interchain region of the immunoglobulin as the N-terminal amino acid; and
(b) a biologically active molecule comprising a functional group capable of reacting with an N terminal cysteine;
wherein the combining of (a) and (b) produces the monomer-dimer hybrid protein.
15. The method of claim 14 , wherein the monomer-dimer hybrid is a dimerically linked monomer-dimer hybrid.
16. The method of claim 14 , wherein the at least one biologically active molecule is chosen from a polypeptide, a nucleic acid molecule, a small organic molecule, a small inorganic molecule.
17. The method of claim 14 , wherein the biologically active molecule is a polypeptide.
18. The method of claim 14 , wherein the functional group is thioester.
19. The methods of claim 18 , wherein the biologically active molecule is a polypeptide comprising an C terminal thioester.
20. The method of claim 14 , wherein the functional group is an aldehyde.
21. The method of claim 14 , wherein both the biologically active molecule and the portion of an immunoglobulin are produced recombinantly.
22. The method of claim 14 , wherein at least one of the biologically active molecule and the portion of an immunoglobulin is produced by chemical synthesis.
23. The method of claim 14 , wherein the chimeric protein further comprises a linker.
24. The method of claim 14 , wherein the reaction is performed in solution.
25. The method of claim 14 , wherein the reaction is performed on a solid support.
26. The method of claim 14 , wherein the biologically active molecule is chosen from EPO, interferon, a viral fusion inhibitor, and a clotting factor.