Antibodies against influenza virus and methods of use thereof

Abstract

The invention provides human scFv antibodies and monoclonal antibodies that neutralize influenza virus. Also provided are methods of treating and/or preventing a influenza related disease or disorder such bird flu The invention also provides methods of vaccinating a patient against influenza. Also provided are methods of diagnosing influenza-related diseases or disorders and methods of detecting the presence of a influenza in a sample.

1 - 75 . (canceled) 
     
     
         76 . An isolated monoclonal antibody, or scFv fragment thereof, wherein said monoclonal antibody is a fully human antibody and wherein said monoclonal antibody binds to an epitope in the stem region of the hemagglutinin (HA) protein of an influenza virus and neutralizes influenza A virus. 
     
     
         77 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 76 , wherein the epitope comprises the amino acid at position 18, 38, 40, and 291 of the HA1 polypeptide. 
     
     
         78 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 77 , wherein the epitope comprises the amino acid at position 18, 19, 20, 21, 38, 41, 42, 45, 49, 52, 53, or 56 of the HA2 polypeptide. 
     
     
         79 . An isolated monoclonal antibody, or an scFv fragment thereof, wherein the monoclonal antibody or scFv fragment thereof, binds to an epitope in the stem region of the hemagglutinin (HA) protein of an influenza virus, wherein the epitope comprises the amino acids at positions 18, 38, 40, and 291 of the HA1 polypeptide. 
     
     
         80 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 79 , wherein the epitope comprises the amino acids at positions 18, 19, 20, 21, and 38 of the HA2 polypeptide. 
     
     
         81 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 79  or  80 , wherein the epitope further comprises the amino acids at positions 41, 42, 45, 49, 52, 53 and 56 of the HA2 polypeptide. 
     
     
         82 . An isolated monoclonal antibody, or scFv fragment thereof, wherein the monoclonal antibody or scFv fragment thereof, binds to the HA protein of an influenza virus and comprises a heavy chain variable domain wherein the heavy chain variable domain comprises a CDR H1, CDR H2, and CDR H3, wherein:
 (a) CDR H1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 44, 45, 46, 47, 48, and 49;   (b) CDR H2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 52, 53, 54, 55, 56, and 57; and   (c) CDR H3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 60, 61, 62, 63, 64, and 65.   
     
     
         83 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 82 , wherein the monoclonal antibody or scFv fragment thereof, further comprises a light chain variable domain wherein the light chain variable domain comprises a CDR L1, CDR L2, and CDRL3, wherein:
 (a) CDR L1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 68, 69, 70, 71, 72, 73, 74, 75, 76, and 77;   (b) CDR L2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 80, 81, 82, 83, 84, 85, 86, 87, 88, and 89; and   (c) CDR L3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 92, 93, 94, 95, 96, 97, 98, 99, 100, and 101.   
     
     
         84 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 82  or  83 , wherein the monoclonal antibody, or scFv fragment thereof, comprises a heavy chain variable domain amino acid sequence selected from the group consisting of SEQ ID NOs: 6, 12, 18, 24, 28, 32, and 36. 
     
     
         85 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 84 , wherein the monoclonal antibody, or scFv fragment thereof, comprises a light chain variable domain amino acid sequence selected from the group consisting of SEQ ID NOs: 8, 10, 14, 16, 20, 22, 26, 30, 34, and 38. 
     
     
         86 . An isolated monoclonal antibody, or scFv fragment thereof, wherein the monoclonal antibody or scFv fragment thereof, binds to the HA protein of an influenza virus and comprises a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises a CDR H1, CDR H2, and CDR H3 of any one of the antibodies provided in  FIG. 3  and wherein the light chain variable domain comprises a CDR L1, CDR L2, and CDR L3 of any one of the antibodies provided in  FIG. 3 . 
     
     
         87 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86 , wherein said monoclonal antibody, or scFv fragment thereof, stabilizes the neutral pH conformation of the HA protein. 
     
     
         88 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86 , wherein said influenza virus is an H1a cluster influenza virus or an H1b cluster influenza virus. 
     
     
         89 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 88 , wherein said influenza virus is an H1, H2, H5, H6, or H11 subtype. 
     
     
         90 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86 , wherein said monoclonal antibody, or scFv fragment thereof, inhibits viral and cell membrane fusion. 
     
     
         91 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86 , wherein said monoclonal antibody, or scFv fragment thereof, competes with the binding of antibody D7, D8, F10, G17, H40, A66, D80, E88, E90, or H98 to the HA protein. 
     
     
         92 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86 , wherein said isolated monoclonal antibody, or scFv fragment thereof, binds to HA with a Kd value of 1 pm-200 mM. 
     
     
         93 . The isolated monoclonal antibody, or scFv fragment thereof, of  claim 92 , wherein said monoclonal antibody, or scFv fragment thereof, binds to HA with a Kd value of about 100-200 pM. 
     
     
         94 . An isolated nucleic acid molecule encoding the isolated monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86 . 
     
     
         95 . The isolated nucleic acid molecule of  claim 94 , wherein the isolated monoclonal antibody, or scFv fragment thereof, comprises a heavy chain variable domain nucleic acid sequence selected from the group consisting of SEQ ID NOs: 5, 7, 13, 15, 21, 23, 29, 33, 37, and 40 and a light chain variable domain nucleic acid sequence selected from the group consisting of SEQ ID NOs: 9, 11, 17, 19, 25, 27, 31, 35, 39, and 42. 
     
     
         96 . A method of treating or preventing a disease or disorder caused by an influenza virus, the method comprising administering to a person suffering from said disease or disorder, or at risk of suffering from said disease or disorder, a therapeutically effective amount of the monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86 . 
     
     
         97 . The method of  claim 96 , wherein the method further comprises administering an antiviral drug, a viral entry inhibitor, or a viral attachment inhibitor. 
     
     
         98 . The method of  claim 97 , wherein said anti-viral drug is a neuraminidase inhibitor, a HA inhibitor, a sialic acid inhibitor, or an M2 ion channel. 
     
     
         99 . The method of  claim 96 , wherein the method comprises administering two or more antibodies specific to a Group I influenza virus. 
     
     
         100 . The method of  claim 96 , wherein the method further comprises administering an antibody specific to a Group II influenza virus. 
     
     
         101 . The method of  claim 96 , wherein said antibody, or scFv fragment thereof, is administered prior to or after exposure to influenza virus. 
     
     
         102 . The method of  claim 96 , wherein said antibody, or scFv fragment thereof, is administered at a dose sufficient to neutralize said influenza virus. 
     
     
         103 . The method of  claim 102 , wherein said influenza virus is an H1, H2, H5, H6, or H11 subtype, or any combinations thereof. 
     
     
         104 . A composition comprising the monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 ,  82 , or  86  and a carrier. 
     
     
         105 . A method of vaccinating a subject against an influenza virus comprising administering to said subject a trimeric hemagglutinin (HA) protein coated or embedded in a biologically compatible matrix. 
     
     
         106 . A method of detecting the presence of an influenza virus in a sample, the method comprising:
 a) contacting the sample with the monoclonal antibody, or scFv fragment thereof, of  claim 76 ,  79 , or  86 ; and   b) detecting the presence or absence of an antibody-antigen complex, thereby detecting the presence of an influenza virus in a sample.   
     
     
         107 . An isolated monoclonal antibody, wherein said monoclonal antibody comprise a VH amino acid sequence encoded by the VH germline gene IGHV 1-69* 01 wherein the amino acid at position: a) 27 is a valine; b) 28 is threonine; c) 30 is serine; d) 31 is serine; e) 54 is methionine; f) 55 is phenylalanine; g) 58 is threonine; h) 100 is proloine; i) 101 is serine; j) 102 is tyrosine; k) 103 is isoleucine and 105 is serine, wherein said monoclonal antibody binds to an epitope in the stem region of the hemagglutinin (HA) protein of an influenza virus, and neutralizes influenza A virus.