Immunoglobulin chimeric monomer-dimer hybrids

Abstract

The invention relates to a chimeric monomer-dimer hybrid protein wherein the protein comprises a first and a second polypeptide chain, the first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and the second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.

1 - 194 . (canceled) 
     
     
         195 . A cell comprising:
 a first nucleic acid construct comprising a first nucleic acid molecule encoding a first polypeptide comprising a biologically active molecule fused to a portion of an immunoglobulin constant region; and   a second nucleic acid construct comprising a second nucleic acid molecule encoding a second polypeptide comprising a portion of an immunoglobulin constant region without a biologically active molecule or an immunoglobulin variable region;   wherein the cell expresses the first polypeptided and/or the second polypeptide when the cell is cultured in a growth medium.   
     
     
         196 . The cell of  claim 195 , wherein the first polypeptide further comprises a linker between the biologically active molecule and the first fragment of the immunoglobulin constant region. 
     
     
         197 . The cell of  claim 196 , wherein the linker is selected form an amino acid sequence set forth in SEQ ID NO: 31, 32, 33, 34, 35, 36, 37, 38, 39, 85, or 86. 
     
     
         198 . The cell of  claim 195 , wherein the first nucleic acid construct and the second nucleic acid construct are viral vectors or plasmids. 
     
     
         199 . The cell of  claim 198 , wherein the viral vectors are chosen from adenovirus vectors, adeno associated virus vectors, murine leukemia virus vectors, and/or mixtures thereof; wherein the plasmids are chosen from pUC, pGGM, pGEX, and/or mixtures thereof. 
     
     
         200 . The cell of  claim 195 , wherein the cell is a prokaryotic or an eukaryotic cell. 
     
     
         201 . The cell of  claim 195 , wherein the portion of the immunoglobulin constant region comprises a Fc neonatal receptor (FcRN) binding partner. 
     
     
         202 . The cell of  claim 195 , wherein the portion of the immunoglobulin constant region comprises a Fc fragment of an IgG, IgA, IgD, IgE, or IgM. 
     
     
         203 . The cell of  claim 195 , wherein the portion of the immunoglobulin constant region comprises a Fc fragment from an IgG1, IgG2, IgG3, or IgG4. 
     
     
         204 . The cell of  claim 195 , wherein the portion of the immunoglobulin constant region comprises an Fc fragment from human IgG1 comprising a FcRn binding partner, and wherein at least one of the Fc fragments in the first polypeptide and in the second polypeptide comprises a N297A mutation. 
     
     
         205 . The cell of  claim 195 , wherein the portion of the immunoglobulin constant region is a peptide mimetic of an Fc fragment of an immunoglobulin. 
     
     
         206 . The cell of  claim 195 , wherein the biologically active molecule is selected from a growth factor or hormone, a viral fusion inhibitor, a clotting factor, a cytokine, a cell surface receptor, analog thereof, or a fragment thereof. 
     
     
         207 . The cell of  claim 195 , wherein the biologically active molecule is an interferon. 
     
     
         208 . A cell comprising:
 a first expression vector containing a first nucleic acid sequence encoding a first polypeptide, wherein the first polypeptide comprises a biologically active molecule fused to a portion of an IgG Fc fragment, optionally via a linker, wherein the IgG Fc fragment includes an FcRn binding site comprising an N297A mutation, and wherein the biologically active molecule is interferon; and   a second expression vector comprising a second nucleic acid sequence encoding a second polypeptide, wherein the second polypeptide comprises a portion of an IgG Fc fragment without a biologically active molecule or an immunoglobulin variable region, and wherein the IgG Fc fragment includes an FcRn binding site comprising an N297A mutation;   wherein the cell expresses the first polypeptide and the second polypeptide when the cell is cultured in a growth medium.   
     
     
         209 . A method for treating a subject with a viral infection comprising:
 administering to the subject a pharmaceutically acceptable amount of a chimeric protein comprising:   a first polypeptide comprising a biologically active molecule fused to a portion of an IgG Fc fragment, optionally via a linker, wherein the IgG Fc fragment includes an FcRn binding site comprising an N297A mutation, and wherein the biologically active molecule is interferon; and   a second polypeptide comprising a portion of an IgG Fc fragment without a biologically active molecule or an immunoglobulin variable region, wherein the IgG Fc fragment includes an FcRn binding site comprising an N297A mutation;   wherein the IgG Fc fragment of the first polypeptide is linked to the IgG Fc fragment of the second polypeptide.   
     
     
         210 . The method of  claim 209 , wherein the biologically active molecule is interferon α or interferon β. 
     
     
         211 . The method of  claim 209 , wherein the biologically active molecule is interferon β and the interferon β is linked to the IgG Fc fragment with a linker. 
     
     
         212 . The method of  claim 211 , wherein the linker is an eight amino acid linker having the amino acid sequence set forth in SEQ ID NO: 31. 
     
     
         213 . The method of  claim 209 , wherein the chimeric protein is formed by the first polypeptide and the second polypeptide obtained from the cell of  claim 208 . 
     
     
         214 . The method of  claim 209 , wherein the chimeric protein is administered to the subject intravenously, subcutaneously, orally, buccally, sublingually, nasally, parenterally, rectally, vaginally, or via a pulmonary route.