title: Newly established monoclonal antibodies for immunological detection of H5N1 influenza virus.
abstract: The H5N1 subtype of the highly pathogenic (HP) avian influenza virus has been  recognized for its ability to cause serious pandemics among humans. In the present study, new  monoclonal antibodies (mAbs) against viral proteins were established for the immunological detection  of H5N1 influenza virus for research and diagnostic purposes. B-cell hybridomas were generated from  mice that had been hyperimmunized with purified A/Vietnam/1194/2004 (NIBRG-14) virion that had been  inactivated by UV-irradiation or formaldehyde. After screening over 4,000 hybridomas, eight  H5N1-specific clones were selected. Six were specific for hemagglutinin (HA) and had in vitro  neutralization activity. Of these, four were able to broadly detect all tested clades of the H5N1  strains. Five HA-specific mAbs detected denatured HA epitope(s) in Western blot analysis, and two  detected HP influenza virus by immunofluorescence and immunohistochemistry. A highly sensitive  antigen-capture sandwich ELISA system was established by combining mAbs with different specificities.  In conclusion, these mAbs may be useful for rapid and specific diagnosis of H5N1 influenza.  Therapeutically, they may have a role in antibody-based treatment of the disease.
authors.affiliation: Department of Immunology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ohnishi
authors.forename: Yoshimasa
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Naoko
authors.initials: N
authors.lastname: Kono
authors.forename: Noriko
authors.initials: N
authors.lastname: Nakajima
authors.forename: Fuminori
authors.initials: F
authors.lastname: Mizukoshi
authors.forename: Shuhei
authors.initials: S
authors.lastname: Misawa
authors.forename: Takuya
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Yu-ya
authors.initials: YY
authors.lastname: Mitsuki
authors.forename: Shu-ichi
authors.initials: S
authors.lastname: Fu
authors.forename: Nakami
authors.initials: N
authors.lastname: Hirayama
authors.forename: Masamichi
authors.initials: M
authors.lastname: Ohshima
authors.forename: Manabu
authors.initials: M
authors.lastname: Ato
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Kageyama
authors.forename: Takato
authors.initials: T
authors.lastname: Odagiri
authors.forename: Masato
authors.initials: M
authors.lastname: Tashiro
authors.forename: Kazuo
authors.initials: K
authors.lastname: Kobayashi
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Itamura
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Tsunetsugu-Yokota
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology / Q000302:isolation & purification; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000918:Antibody Specificity; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology; D005260:Female; D005455:Fluorescent Antibody Technique; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007150:Immunohistochemistry; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D008168:Lung / Q000276:immunology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D038301:Virus Inactivation
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000939:Epitopes; D019267:Hemagglutinin Glycoproteins, Influenza Virus