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 . An isolated monoclonal antibody or scFv antibody, wherein said antibody binds to an epitope in the stem region of the hemagglutinin (HA) protein of an influenza virus, and neutralizes influenza A virus, wherein the epitope comprises the amino acid at position 18, 38, 40, 291 of the HA1 polypeptide and the amino acid at position 18, 19, 20, 21, 38, 41, 42, 45, 49, 52, 53 and 56 of the HA2 polypeptide. 
     
     
         2 . The antibody of  claim 1 , wherein said influenza virus is a Group I influenza virus. 
     
     
         3 . The antibody of  claim 1 , wherein said influenza virus is H1 cluster influenza virus. 
     
     
         4 . The antibody of  claim 3 , wherein said HI cluster influenza virus is an H1a cluster influenza virus or an H1b cluster influenza virus. 
     
     
         5 . The antibody of  claim 1 , wherein said epitope is non-linear. 
     
     
         6 . The monoclonal antibody of  claim 1 , wherein said epitope comprises the HA1 and the HA2 polypeptide. 
     
     
         7 . (canceled) 
     
     
         8 . The antibody of  claim 1 , wherein said antibody inhibits viral and cell membrane fusion. 
     
     
         9 . The antibody of  claim 1 , wherein said antibody competes with the binding of monoclonal antibody D7, D8, F10, G17, H40, A66, D80, E88, E90, or H98 to the HA protein. 
     
     
         10 . The antibody of  claim 1  wherein said antibody is a fully human antibody. 
     
     
         11 .- 19 . (canceled) 
     
     
         20 . A method of treating or preventing a disease or disorder caused by an influenza virus, the method comprising administering to a person at risk of suffering from said disease or disorder, a therapeutically effective amount of the antibody of  claim 1 . 
     
     
         21 . The method of  claim 20 , wherein the method further comprises administering an antiviral drug, a viral entry inhibitor or a viral attachment inhibitor. 
     
     
         22 . The method of  claim 21 , wherein said anti-viral drug is a neuraminidase inhibitor, a HA inhibitor, a sialic acid inhibitor or an M2 ion channel. 
     
     
         23 . The method of  claim 22  wherein said M2 ion channel inhibitor is amantadine or rimantadine. 
     
     
         24 . The method of  claim 22 , wherein said neuraminidase inhibitor is zanamivir, or oseltamivir phosphate. 
     
     
         25 . The method of  claim 20 , wherein the method comprises administering two or more antibodies specific to a Group I influenza virus. 
     
     
         26 . The method of  claim 20 , wherein the method further comprises administering an antibody specific to a Group II influenza virus. 
     
     
         27 . The method of  claim 20 , wherein said antibody is administered prior to or after exposure to influenza virus. 
     
     
         28 . The method of  claim 20 , wherein said antibody is administered at a dose sufficient to neutralize said influenza virus. 
     
     
         29 .- 37 . (canceled) 
     
     
         38 . A composition comprising the antibody of  claim 1  and a carrier. 
     
     
         39 . The composition of  claim 38 , further comprising an anti-viral drug, a viral entry inhibitor or a viral attachment inhibitor. 
     
     
         40 . The composition of  claim 39 , wherein said anti-viral drug is a neuraminidase inhibitor, a HA inhibitor, a sialic acid inhibitor or an M2 ion channel. 
     
     
         41 . The composition of  claim 40 , wherein said M2 ion channel inhibitor is amantadine or rimantadine. 
     
     
         42 . The composition of  claim 40 , wherein said neuraminidase inhibitor zanamivir, or oseltamivir phosphate. 
     
     
         43 . The composition of  claim 38 , further comprising one or more antibodies specific to a Group I influenza virus 
     
     
         44 . The composition of  claim 38 , further comprising an antibody specific to a Group II influenza virus. 
     
     
         45 . A method identifying an antibody specific for an influenza virus comprising:
 contacting a trimeric hemagglutinin (HA) protein, wherein said HA protein is immobilized on a solid surface with a test antibody and detecting a HA protein-antibody complex.   
     
     
         46 . (canceled) 
     
     
         47 . The method of  claim 45 , wherein said HA protein is produced in a non-mammalian cell. 
     
     
         48 . The method of  claim 45 , wherein said HA protein is produced in insect cells. 
     
     
         49 . The method of  claim 45 , wherein said HA protein comprising modified glycosylations compared to a wild type HA protein. 
     
     
         50 . The method of  claim 45  wherein said HA protein is immobilized on said solid surface as to mask the globular head of said HA protein. 
     
     
         51 . A method of producing an isolated antibody that specifically binds a trimeric hemagglutinin (HA) protein, (a) exposing a single chain or Fab expression library to said HA protein wherein said HA protein is immobilized on a solid surface; (b) identifying an antibody in said library that specifically binds said protein: and (c) isolating the antibody from the library. 
     
     
         52 . (canceled) 
     
     
         53 . The method of  claim 51 , wherein said HA protein is produced in a non-mammalian cell. 
     
     
         54 . The method of  claim 51 , wherein said HA protein comprising modified glycosylations compared to a wild type HA protein. 
     
     
         55 . The method of  claim 51 , wherein said HA protein is immobilized on said solid surface as to mask the globular head of said HA protein. 
     
     
         56 . The method of  claim 51  wherein said expression library is a non-immune library or an H5 naïve library. 
     
     
         57 . (canceled) 
     
     
         58 . A method of vaccinating a subject against an influenza virus comprising administering to said subject an trimeric hemagglutinin (HA) protein coated or embedded in a biologically compatible matrix. 
     
     
         59 . The method of  claim 58 , wherein said HA protein has non-mammalian carbohydrate structures. 
     
     
         60 . The method of  claim 58 , wherein said HA protein comprising modified glycosylations compared to a wild type HA protein. 
     
     
         61 . The method of  claim 58 , wherein said HA protein is coated or embedded in said biologically compatible matrix as to mask the globular head of said HA protein. 
     
     
         62 . A method of producing an isolated antibody that specifically binds a pathogenic enveloped virus, (a) exposing a single chain or Fab expression library to a membrane fusion protein of said virus wherein said protein is immobilized on a solid surface; (b) identifying an antibody in said library that specifically binds said protein; and (c) isolating the antibody from the library. 
     
     
         63 . (canceled) 
     
     
         64 . The method of  claim 62 , wherein said membrane fusion protein is produced in a nonmammalian cell. 
     
     
         65 . The method of  claim 62 , wherein said membrane fusion protein has non-mammalian carbohydrate structures. 
     
     
         66 . The method of  claim 62 , wherein said membrane fusion protein comprising modified glycosylations compared to a wild type membrane fusion protein. 
     
     
         67 . The method of  claim 62 , wherein said expression library is a non-immune library or an H5 naïve library. 
     
     
         68 . (canceled) 
     
     
         69 . A method of vaccinating a subject against pathogenic enveloped virus comprising administering to said subject a membrane fusion protein of said virus coated or embedded in a biologically compatible matrix. 
     
     
         70 . The method of  claim 69 , wherein said fusion protein has non-mammalian carbohydrate structures. 
     
     
         71 . The method of  claim 69 , wherein said fusion protein comprising modified glycosylations compared to a wild type membrane fusion protein. 
     
     
         72 . A method of detecting the presence of an influenza virus in a sample, the method comprising:
 a) contacting the sample with the monoclonal antibody of claim; and   b) detecting the presence or absence of an antibody-antigen complex,   thereby detecting the presence of an influenza virus in a sample.   
     
     
         73 . The method of  claim 72 , wherein the detecting occurs in vivo. 
     
     
         74 . (canceled) 
     
     
         75 . An isolated monoclonal antibody, wherein said monoclonal antibody comprise a VH amino acid sequence encoded by the VH germline gene IGHVI-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 proline; i) 101 is serine; j) 102 is tyrosine; k) 10 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.