NEUTRALIZING MOLECULES TO VIRAL ANTIGENS
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.
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.