Recombinant Human Albumin-Granulocyte Macrophage Colony Stimulating Factor Fusion Protein With Long-Lasting Biological Effects

Abstract

Compositions, kits and methods are provided for promoting general health or for prevention or treatment of diseases by using novel recombinant fusion proteins of human serum albumin (HSA) and bioactive molecules. The bioactive molecules may be a protein or peptide having a biological function in vitro or in vivo, and preferably, having a therapeutic activity when administered to a human. By fusing the bioactive molecule to HSA, stability of the bioactive molecule in vivo can be improved and the therapeutic index increased due to reduced toxicity and longer-lasting therapeutic effects in vivo. In addition, manufacturing processes are provided for efficient, cost-effective production of these recombinant proteins in yeast.

1 . A human serum albumin-granulocyte macrophage colony stimulating factor (HSA-GM-CSF) fusion protein comprising: (a) the amino acid sequence set forth in SEQ ID NO: 10; (b) the amino acid sequence encoded by the polynucleotide set forth in SEQ ID NO: 9; or (c) the amino acid sequence encoded by the polynucleotide contained in the yeast strain designated as YZ-HSA/hGM-CSF in ATCC® Deposit No: PTA-4607, said HSA-GM-CSF fusion protein having a plasma half-life longer than a plasma half-life of granulocyte macrophage colony stimulating factor. 
     
     
         2 . The HSA-GM-CSF fusion protein of  claim 1 , wherein said HSA-GM-CSF fusion protein has a shelf-life four times longer than a shelf-life of the granulocyte macrophage colony stimulating factor. 
     
     
         3 . The HSA-GM-CSF fusion protein of  claim 1 , wherein said HSA-GM-CSF fusion protein has said plasma half-life four times longer than said plasma half-life of the granulocyte macrophage colony stimulating factor. 
     
     
         4 . A composition comprising said HSA-GM-CSF fusion protein of  claim 1  and a pharmaceutically acceptable excipient. 
     
     
         5 . The composition of  claim 4  further comprising a human serum albumin-cell proliferation stimulatory factor (HSA-CPSF) fusion protein different from said HSA-GM-CSF fusion protein. 
     
     
         6 . A polynucleotide comprising a polynucleotide sequence encoding said HSA-GM-CSF fusion protein of  claim 1 . 
     
     
         7 . A vector comprising said polynucleotide of  claim 6 . 
     
     
         8 . The vector of  claim 7 , wherein said vector is an expression vector for expressing said fusion protein in a host organism comprising mammal, fish, insect, plant, yeast, or bacterium. 
     
     
         9 . The vector of  claim 8 , wherein said host organism is said yeast. 
     
     
         10 . The vector of  claim 9 , wherein a strain of said yeast is selected from the group consisting of  Saccharomyces, Candida, Pichia, Kluyveromyces, Torulaspora , and  Schinosaccharomyces.    
     
     
         11 . The vector of  claim 9 , wherein a strain of said yeast is  Pichia pastoris.    
     
     
         12 . The vector of  claim 7 , wherein said vector is a yeast transfer vector comprising pPICZ A, pPICZ B, or pPICZ C. 
     
     
         13 . A host cell comprising said vector of  claim 7 . 
     
     
         14 . The host cell of  claim 13 , wherein said host cell is selected from the group consisting of mammalian, fish, insect, plant, yeast, and bacterial cells. 
     
     
         15 . The host cell of  claim 14 , wherein said mammalian cells are Chinese hamster ovarian (CHO) cells.