Methods for Chemically Synthesizing Immunoglobulin Chimeric Proteins

Abstract

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.