Newly Established Monoclonal Antibodies for Immunological Detection of H5N1 Influenza Virus

作者：Ohnishi Kazuo; Takahashi Yoshimasa; Kono Naoko; Nakajima Noriko; Mizukoshi Fuminori; Misawa Shuhei; Yamamoto Takuya; Mitsuki Yu ya; Fu Shu ichi; Hirayama Nakami; Ohshima Masamichi; Ato Manabu; Kageyama Tsutomu; Odagiri Takato; Tashiro Masato; Kobayashi Kazuo; Itamura Shigeyuki; Tsunetsugu Yokota Yasuko^*

来源：Japanese Journal of Infectious Diseases, 2012, 65(1): 19-27.

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.

• 出版日期2012-1