title: Generation and Characterization of Monoclonal Antibodies Specific to Avian Influenza H5N1 Hemagglutinin Protein.
abstract: Highly pathogenic avian influenza (HPAI) H5N1 virus has in the past breached the species barrier from infected domestic poultry to humans in close contact. Although human-to-human transmission has previously not been reported, HPAI H5N1 virus has pandemic potential owing to gain of function mutation(s) and/or genetic reassortment with human influenza A viruses. Monoclonal antibodies (MAbs) have been used for diagnosis as well as specific therapeutic candidates in several disease conditions including viral infections in humans. In this study, we describe the preliminary characterization of four murine MAbs developed against recombinant hemagglutinin (rHA) protein of avian H5N1 A/turkey/Turkey/1/2005 virus that are either highly specific or broadly reactive against HA from other H5N1 subtype viruses, such as A/Hong Kong/213/03, A/Common magpie/Hong Kong/2256/2006, and A/Barheaded goose/Quinghai/14/2008. The antibody binding is specific to H5N1 HAs, as none of the antibodies bound H1N1, H2N2, H3N2, or B/Brisbane/60/2008 HAs. Out of the four MAbs, one of them (MA-7) also reacted weakly with the rHA protein of H7N9 A/Anhui/1/2013. All four MAbs bound H5 HA (A/turkey/Turkey/1/2005) with high affinity with an equilibrium dissociation constant (KD) ranging between 0.05 and 10.30 nM. One of the MAbs (MA-1) also showed hemagglutination inhibition activity (HI titer; 31.25 μg/mL) against the homologous A/turkey/Turkey/1/2005 H5N1 virus. These antibodies may be useful in developing diagnostic tools for detection of influenza H5N1 virus infection.
authors.affiliation: 1 Reproductive Cell Biology Lab, National Institute of Immunology , New Delhi, India .
authors.forename: Ankita
authors.initials: A
authors.lastname: Malik
authors.affiliation: 2 Molecular Biophysics Unit, Indian Institute of Science , Bangalore, India .
authors.forename: V Vamsee Aditya
authors.initials: VV
authors.lastname: Mallajosyula
authors.affiliation: 1 Reproductive Cell Biology Lab, National Institute of Immunology , New Delhi, India .
authors.forename: Nripendra Nath
authors.initials: NN
authors.lastname: Mishra
authors.affiliation: 2 Molecular Biophysics Unit, Indian Institute of Science , Bangalore, India .
authors.forename: Raghavan
authors.initials: R
authors.lastname: Varadarajan
authors.affiliation: 1 Reproductive Cell Biology Lab, National Institute of Immunology , New Delhi, India .
authors.forename: Satish Kumar
authors.initials: SK
authors.lastname: Gupta
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000914:Antibodies, Viral / Q000096:biosynthesis*; D000918:Antibody Specificity; D001201:Ascites / Q000276:immunology; D003429:Cross Reactions; D005260:Female; D006385:Hemagglutination Inhibition Tests; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000008:administration & dosage / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000737:chemistry / Q000276:immunology; D053122:Influenza A Virus, H3N2 Subtype / Q000737:chemistry / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000737:chemistry / Q000276:immunology*; D064766:Influenza A Virus, H7N9 Subtype / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000032:analysis / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; C483468:hemagglutinin, human influenza A virus