Production and characterization of monoclonal antibodies of H5 avian influenza virus

• Main Authors: Tung-Hau Li, 李同浩
• Other Authors: Happy K. Shieh
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/99595812520358884111

碩士 === 中興大學 === 獸醫微生物學研究所 === 95 === The outbreaks of highly pathogenic H5N1 influenza A virus from 1997 to the present day，The virus cancause economical losses to the poultry industry and have devastating consequences for animal health，H5N1 infections have resulted in 318 laboratory-confirmed infected people，including 192 deaths。HPAI receives immense attention in the veterinary world and is globally treated as a disease immediately notifiable on suspicion to the authorities。The purpose of this study is to prepare monoclonal antibodies against H5N3 AIV and to use these monoclonal antibodies to develop a rapid method for detection of AIV。BALB/c mice were immunized with H5N3 virus and then the splenocytes from the immunized mouse were collected and fused with NS-1 myeloma cells。 The hybridomas secreting antibodies against AIV were screened by hemagglutination inhibition、dot immunoblotting、Western blotting and immunofluorescence assay。Two hybridomas which secreted specific antibodies to AIV，were selected among 3000 hybridomas in five fusion experiments。Cloning of these established hybridomas was carried by the limiting dilution method in order to confirm their monoclonality。After two times cloning，two monoclonal antibodies were purified.Hybridomas C2-8G and D1-11G，secreting IgA and IgG2a respectively。