Methods for Chemically Synthesizing Immunoglobulin Chimeric Proteins
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 at least one biologically active molecule and at least a portion of an immunoglobulin constant region, wherein
a) the portion of an immunoglobulin constant region comprises an N terminus cysteine and
b) the biologically active molecule comprises a functional group capable of reacting with an N terminus cysteine to form a bond.
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 at least one biologically active molecule is chosen from a polypeptide, a nucleic acid molecule, a small organic molecule, a small inorganic molecule.
5 . (canceled)
6 . (canceled)
7 . The method of claim 1 , wherein the functional group is thioester.
8 . (canceled)
9 . The method of claim 1 , wherein the functional group is an aldehyde.
10 . The method of claim 1 , wherein both the biologically active molecule and the portion of an immunoglobulin are produced recombinantly.
11 . 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.
12 . The method of claim 1 , wherein the portion of an immunoglobulin is an FcRn binding partner.
13 . The method of claim 12 , the FcRn binding partner is an Fc or a fragment thereof.
14 . (canceled)
15 . (canceled)
16 . (canceled)
17 . The method of claim 1 , wherein the biologically active molecule is chosen from EPO, interferon, a viral fusion inhibitor, and a clotting factor.
18 . A method of synthesizing a chimeric protein comprising
a) recombinantly expressing a fusion protein comprising at least a portion of an immunoglobulin constant region and a splicing protein capable of forming a C terminus thioester on the portion of an immunoglobulin constant region;
b) adding a thiol cofactor to the fusion protein of a);
c) adding at least one biologically active molecule having an N terminal cysteine, thereby synthesizing the chimeric protein.
19 . The method of claim 18 , wherein the splicing protein is intein.
20 . The method of claim 18 , wherein the thiol cofactor is MESNA.
21 . The method of claim 18 , wherein the chimeric protein is chosen from a dimer and a monomer-dimer hybrid.
22 . The method of claim 21 , wherein the monomer-dimer hybrid is a dimerically linked monomer-dimer hybrid.
23 . The method of claim 18 , 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.
24 . (canceled)
25 . (canceled)
26 . The method of claim 18 , wherein both the biologically active molecule and the portion of an immunoglobulin are produced recombinantly.
27 . The method of claim 18 , wherein at least one of the biologically active molecule and the portion of an immunoglobulin is produced by chemical synthesis.
28 . The method of claim 18 , wherein the portion of an immunoglobulin is an FcRn binding partner.
29 . The method of claim 28 , the FcRn binding partner is an Fc or a fragment thereof.
30 . (canceled)
31 . (canceled)
32 . (canceled)
33 . (canceled)
34 . The method of claim 18 , wherein the biologically active molecule is chosen from EPO, interferon, a viral fusion inhibitor, and a clotting factor.