US2015232516A1PendingUtilityA1

Flagellin Fusion Proteins

62
Assignee: VAXINNATE CORPPriority: Mar 7, 2006Filed: Apr 30, 2015Published: Aug 20, 2015
Est. expiryMar 7, 2026(expired)· nominal 20-yr term from priority
Inventors:Langzhou SongValerian NakaarAlbert E. PriceLynda G. TusseyJames W. HuleattThomas J. PowellRobert K. Evans
A61K 2039/6068A61K 39/0258C12N 2770/24122C12N 2760/16271C12N 15/62C07K 14/11C07K 2319/00A61K 2039/575A61P 31/16A61P 31/00A61P 37/04A61K 39/385C12N 2760/16134A61K 2039/6037C07K 14/195C07K 14/005C12N 2760/16222C12N 2760/16171C12N 2770/24134C07K 14/255C12N 7/00C12N 2760/16122A61K 39/145A61K 39/02C12N 2760/16334A61K 39/12A61K 47/42A61K 2039/6018C12N 2760/16234C12N 2760/16322A61K 2039/543A61K 2039/6075A61K 39/00Y02A50/30
62
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Claims

Abstract

Methods of making a protein that stimulates a protective immune response in a subject include separating a portion of a protein from a naturally occurring influenza viral hemagglutinin to form a protein portion. The protein portion includes at least a portion of a globular head, and at least a portion of at least one secondary structure having at least one β-sheet at a bottom of the globular head that causes the globular head to essentially retain its tertiary structure. The protein portion made by the methods of the invention lacks a transmembrane domain, a cytoplasmic domain and an HA2 subunit. A nucleic acid sequence encoding the protein portion is transformed into a prokaryotic host cell.

Claims

exact text as granted — not AI-modified
1 . (canceled) 
     
     
         2 . A fusion protein comprising at least a portion of a flagellin protein that initiates an intracellular signal transduction pathway for a Toll-like Receptor and at least a portion of at least one an influenza viral antigen inserted into a loop of a domain 3 of the flagellin protein. 
     
     
         3 . The fusion protein of  claim 2 , wherein the fusion protein is incorporated into a liposome. 
     
     
         4 . The fusion protein of  claim 2 , wherein the fusion protein is attached to a support. 
     
     
         5 . The fusion protein of  claim 4 , wherein the support is a polymer bead. 
     
     
         6 . The fusion protein of  claim 4 , wherein the support is a virus. 
     
     
         7 . The fusion protein of  claim 4 , wherein the support is a nucleic acid. 
     
     
         8 . The fusion protein of  claim 4 , wherein the support is an adjuvant. 
     
     
         9 . The fusion protein of  claim 2 , wherein the flagellin protein includes at least one member selected from the group consisting of a  Salmonella typhimurium  flagellin protein, an  E. coli  flagellin protein, a  Salmonella muenchen  flagellin protein, a  Yersinia  flagellin protein, a  P. aeruginosa  flagellin protein and a  Listeria monocytogenes  flagellin protein. 
     
     
         10 . The fusion protein of  claim 9 , wherein the flagellin protein is the  Salmonella typhimurium  flagellin protein. 
     
     
         11 . The fusion protein of  claim 10 , wherein the  Salmonella typhimurium  flagellin protein is SEQ ID NO: 841. 
     
     
         12 . The fusion protein of  claim 2 , wherein the portion of the influenza viral antigen includes an HA1-1 portion of an influenza viral hemagglutinin antigen. 
     
     
         13 . The fusion protein of  claim 2 , wherein the portion of the influenza viral antigen includes an HA1-2 portion of an influenza viral hemagglutinin antigen. 
     
     
         14 . The fusion protein of  claim 2 , wherein the influenza viral antigen is at least a portion of an influenza B viral antigen. 
     
     
         15 . The fusion protein of  claim 14 , wherein the influenza B viral antigen is inserted into the loop of the domain 3 of the flagellin protein defined by amino acid residues 196-216 of SEQ ID NO: 841. 
     
     
         16 . The fusion protein of  claim 14 , wherein the influenza B viral antigen is inserted into the loop of the domain 3 of the flagellin protein defined by amino acid residues 259-274 of SEQ ID NO: 841. 
     
     
         17 . The fusion protein of  claim 14 , wherein the influenza B viral antigen is inserted into the loop of the domain 3 of the flagellin protein defined by amino acid residues 266-281 of SEQ ID NO: 841. 
     
     
         18 . The fusion protein of  claim 14 , wherein the influenza B viral antigen is inserted into the loop of the domain 3 of the flagellin protein defined by amino acid residues 244-248 of SEQ ID NO: 841. 
     
     
         19 . The fusion protein of  claim 14 , wherein the influenza B viral antigen is at least a portion of an influenza B viral hemagglutinin antigen. 
     
     
         20 . The fusion protein of  claim 19 , wherein the portion of the influenza B viral hemagglutinin antigen lacks a transmembrane domain and a cytoplasmic domain. 
     
     
         21 . The fusion protein of  claim 20 , wherein the portion of the influenza B viral hemagglutinin antigen includes at least a portion of a globular head. 
     
     
         22 . The fusion protein of  claim 21 , wherein the portion of the influenza B viral hemagglutinin antigen further lacks an HA2 subunit. 
     
     
         23 . The fusion protein of  claim 21 , wherein the portion of the globular head includes at least one β-sheet at a bottom of the portion of the globular head. 
     
     
         24 . The fusion protein of  claim 23 , wherein the fusion protein is incorporated into a liposome. 
     
     
         25 . The fusion protein of  claim 23 , wherein the fusion protein is attached to a support. 
     
     
         26 . The fusion protein of  claim 25 , wherein the support is a polymer bead. 
     
     
         27 . The fusion protein of  claim 25 , wherein the support is a virus. 
     
     
         28 . The fusion protein of  claim 25 , wherein the support is a nucleic acid. 
     
     
         29 . The fusion protein of  claim 25 , wherein the support is an adjuvant. 
     
     
         30 . The fusion protein of  claim 23 , wherein the portion of the globular head further includes at least one β-sandwich at the bottom of the portion of the globular head. 
     
     
         31 . The fusion protein of  claim 30 , wherein the portion of the globular head further includes at least two β-strands at the bottom of the portion of the globular head. 
     
     
         32 . The fusion protein of  claim 2 , further including a linker between at least one of an amino-terminus or a carboxy-terminus of the influenza viral antigen and the loop of the domain 3 of the flagellin protein. 
     
     
         33 . The fusion protein of  claim 32 , wherein the linker is an amino acid linker. 
     
     
         34 . The fusion protein of  claim 2 , further including an additional antigen fused to the flagellin protein at a site that is distinct from insertion of the influenza viral antigen into the loop of the domain 3 of the flagellin protein. 
     
     
         35 . The fusion protein of  claim 34 , wherein the additional antigen is at least one member selected from the group consisting of an influenza A viral antigen and an influenza B viral antigen. 
     
     
         36 . The fusion protein of  claim 35 , wherein the additional antigen is the influenza A viral antigen. 
     
     
         37 . The fusion protein of  claim 36 , wherein the influenza A viral antigen is an influenza A hemagglutinin influenza A viral antigen. 
     
     
         38 . The fusion protein of  claim 34 , further including a linker between at least one of an amino-terminus or a carboxy-terminus of the additional antigen and the flagellin protein. 
     
     
         39 . The fusion protein of  claim 38 , wherein the linker is an amino acid linker.

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