Production of flu vaccine in myceliophthora thermophila

Abstract

Recombinant expression of influenza virus surface proteins in the fungus Myceliophthora thermophila strain C1 is provided. The recombinant proteins are for use in influenza vaccine compositions.

1 . A substantially pure, recombinant influenza virus surface protein produced by a genetically modified Thermothelomyces thermophila C1, wherein the influenza virus surface protein is purified to 95% purity or greater and is active and immunogenic, and induces a protective immune response when used as a vaccine. 
     
     
         2 . The influenza protein of  claim 1 , wherein the influenza virus surface protein is hemagglutinin (HA). 
     
     
         3 . The influenza protein of  claim 2 , wherein the HA subtype is selected from the group consisting of influenza A-H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15 and H16; and influenza B subtype. 
     
     
         4 . The influenza protein of  claim 2 , wherein the HA subtype is a subtype infecting humans selected from the influenza A subtypes H1, H2 and H3; and influenza B subtype. 
     
     
         5 . The influenza protein of  claim 2 , wherein the HA is from an influenza virus strain selected from the group consisting of A/New Caledonia /20/99 (H1N1), A/Califomia/04/2009 (H1N1), A/Uruguay/716/07 (H3N2) (A/Brisbane/10/07-like), B/Florida/04/2006: B Yamagata lineage, A/Puerto Rico/08/1934 (H1N1), and A/Texas/50/2012 (H3N2). 
     
     
         6 . The influenza protein of  claim 1 , wherein the influenza virus surface protein is a neuraminidase (NA). 
     
     
         7 . The influenza protein of  claim 6 , wherein the NA subtype is selected from the group consisting of influenza A- N1, N2, N3, N4, N5, N6, N7, N8 and N9; and influenza B subtype. 
     
     
         8 . The influenza protein of  claim 6 , wherein the NA subtype is a subtype infecting humans selected from influenza A subtypes N1 and N2; and influenza B subtype. 
     
     
         9 . An influenza vaccine composition comprising the influenza virus surface protein of  claim 1 . 
     
     
         10 . The influenza vaccine composition of  claim 9 , formulated for parenteral administration. 
     
     
         11 . The influenza vaccine composition of  claim 9 , formulated for intramuscular administration, intradermal administration, mucosal delivery, oral administration, or intranasal delivery. 
     
     
         12 . The influenza protein of  claim 1 , wherein the genetically-modified  Thermothelomyces thermophila  C1 comprising an expression construct comprising a nucleic acid sequence encoding the influenza virus surface protein operably linked to at least one C1 regulatory sequence, wherein the influenza virus surface protein comprises its ectodomain and transmembrane domain and is expressed in the C1 as a membrane-bound protein. 
     
     
         13 . The influenza protein of  claim 1 , wherein the genetically-modified  Thermothelomyces thermophila  C1 comprising an expression construct comprising a nucleic acid sequence encoding the influenza virus protein operably linked to at least one C1 regulatory sequence comprising a C1 promoter, and wherein the expression construct further comprising a C1 signal peptide. 
     
     
         14 . A method for producing an influenza virus protein, the method comprising culturing a  Thermothelomyces thermophila  C1 under conditions suitable for expressing the influenza virus protein; and recovering the influenza virus protein, wherein the  Thermothelomyces thermophila  C1 comprises an expression construct comprising a nucleic acid sequence encoding the influenza virus protein operably linked to at least one C1 regulatory sequence comprising a C1 promoter, and wherein the expression construct further comprising a C1 signal peptide.