Neutralizing molecules to viral antigens

Abstract

The present invention concerns methods and means for identifying, producing, and engineering neutralizing molecules against influenza A viruses, and to the neutralizing molecules produced. In particular, the invention concerns neutralizing molecules against various influenza A virus subtypes, including neutralizing antibodies against H5 and/or H3 and/or H1, such as, for example all of H1, H3, and H5 subtypes, and methods and means for making such molecules.

What is claimed is: 
     
         1 . A vaccine effective against an influenza A virus comprising a polypeptide that comprises a hemagglutinin (HA) stem polypeptide or fragment thereof. 
     
     
         2 . A vaccine effective against an influenza A virus comprising a polypeptide that functionally mimics a neutralization epitope of a molecule that (i) neutralizes more than one subtype and/or more than one isolate of an influenza A virus, (ii) binds to a hemagglutinin (HA) antigen of the virus, and (iii) does not inhibit hemagglutination, wherein the polypeptide comprises a hemagglutinin (HA) stem polypeptide or fragment thereof. 
     
     
         3 . The vaccine of  claim 1  or  2 , wherein the polypeptide comprises an HA globular head region polypeptide or fragment thereof. 
     
     
         4 . The vaccine of  claim 1  or  2 , wherein the polypeptide lacks an HA globular head polypeptide or fragment thereof. 
     
     
         5 . The vaccine of  claim 1  or  2 , wherein the polypeptide lacks an HA1 globular head polypeptide. 
     
     
         6 . The vaccine of  claim 1  or  2 , wherein the polypeptide lacks a hemagglutinin binding domain. 
     
     
         7 . The vaccine of  claim 1  or  2 , effective against more than one isolate of an influenza A virus subtype and/or more than one subtype of the influenza A virus. 
     
     
         8 . The vaccine of  claim 7 , effective against at least two influenza A virus subtypes selected from the group consisting of H1, H2, H3, H5, H6, H7, H8, and H9. 
     
     
         9 . The vaccine of  claim 7 , effective against at least two isolates of an influenza A virus subtype selected from the group consisting of H1, H2, H3, H5, H6, H7, H8, and H9. 
     
     
         10 . The vaccine of  claim 7 , effective against an H1 influenza A virus subtype. 
     
     
         11 . The vaccine of  claim 7 , effective against at least one isolate of H1 influenza A virus subtype. 
     
     
         12 . The vaccine of  claim 7 , effective against H5 influenza A virus subtype. 
     
     
         13 . The vaccine of  claim 7 , effective against at least one isolate of H5 influenza A virus subtype. 
     
     
         14 . The vaccine of  claim 7 , effective against H1 and H5 influenza A virus subtypes. 
     
     
         15 . The vaccine of  claim 7 , effective against isolates of H1 and H5 influenza A virus subtypes. 
     
     
         16 . The vaccine of  claim 2 , wherein said molecule is an antibody. 
     
     
         17 . The vaccine of  claim 16 , wherein the antibody is an antibody which binds essentially the same epitope as the epitope for an antibody comprising a heavy chain polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:45, SEQ ID NO:9, and SEQ ID NO:61; or a consensus or variant sequence based upon said amino acid sequences, or a fragment thereof. 
     
     
         18 . The vaccine of  claim 16 , wherein the antibody is an antibody comprising a heavy chain polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:45, SEQ ID NO:9, and SEQ ID NO:61; or a consensus or variant sequence based upon said amino acid sequences, or a fragment thereof. 
     
     
         19 . The vaccine of  claim 16 , wherein the antibody is an antibody which binds essentially the same epitope as the epitope for an antibody comprising a heavy chain polypeptide comprising an amino acid sequence having the formula: M-A-X 1 -X 2 -Q-L-V-Q-S-G-X 3 -E-V-X 4 -K-P-G-X 5 -S-V-X 6 -X 7 -S-C-K-X 8 -S-G-G-X 9 -F-S-S-Y-A-X 10 -X 11 -W-V-R-Q-A-P-G-W-M-G-X 12 -G-I-I-X 13 -X 14 -F-G-T-T-X 15 -N-Y-A-Q-K-F-Q-G-R-X 16 -T-X 17 -T-A-D-X 18 -X 19 -T-S-T-A-Y-M-E-L-S-S-L-R-S-X 20 -D-T-A-V-Y-Y-C-A-R-G-S-Y-Y-Y-E-X 21 -X 22 -L-D-Y-W-G-X 23 -G-T-X 24 -V-T-V-S-S; or a consensus or variant sequence based upon said amino acid sequences, or a fragment thereof; wherein X 1  is Q or E; X 2  is V or M; X 3  is A or T; X 4  is K or Q; X 5  is S or A; X 6  is K or R; X 7  is V or L; X 8  is A, T or V; X 9  is T, S or A; X 10  is I or V; X II  is S or T; X 12  is G or A; X 13  is P or G; X 14  is I or M; X 15  is A or T; X 16  is V or L; X 17  is I, L, or M; X 18  is K or E; X 19  is S, L or M; X 20  is E or D; X 21  is S, T or N; X 22  is S or T; X 23  is Q, K, G or R; and X 24  is L, T or M. 
     
     
         20 . The vaccine of  claim 16 , wherein the antibody is an antibody comprising a heavy chain polypeptide comprising an amino acid sequence having the formula: M-A-X 1 -X 2 -Q-L-V-Q-S-G-X 3 -E-V-X 4 -K-P-G-X 5 -S-V-X 6 -X 7 -S-C-K-X 8 -S-G-G-X 9 F-S-S-Y-A-X 10 -X 11 -W-V-R-Q-A-P-G-Q-G-L-E-W-M-G-X 12 -G-I-I-X 13 -X 14 -F-G-T-T-X 15 -N-Y-A-Q-K-F-Q-G-R-X 16 -T-X 17 -T-A-D-X 18 -X 19 -T-S-T-A-Y-M-E-L-S-S-L-R-S-X 20 -D-T-A-V-Y-Y-C-A-R-G-S-Y-Y-Y-E-X 21 -X 22 -L-D-Y-W-G-X 23 -G-T-X 24 -V-T-V-S-S; or a consensus or variant sequence based upon said amino acid sequences, or a fragment thereof; wherein X 1  is Q or E; X 2  is V or M; X 3  is A or T; X 4  is K or Q; X 5  is S or A; X 6  is K or R; X 7  is V or L; X 8  is A, T or V; X 9  is T, S or A; X 10  is I or V; X 11  is S or T; X 12  is G or A; X 13  is P or G; X 14  is I or M; X 15  is A or T; X 16  is V or L; X 17  is I, L, or M; X 18  is K or E; X 19  is S, L or M; X 20  is E or D; X 21  is S, T or N; X 22  is S or T; X 23  is Q, K, G or R; and X 24  is L, T or M.