Preparation Of Monoclonal Antibodies Against Hemagglutinin Of Avian Influenza Virus Subtype H5 And Establishment Of Double Antibodies Sandwich ELISA

Inoculate subtype H5 AIV inactivated to 6～8 week-old Balb/c mice. By using the hybridoma technique, SP2/0 myeloma cells and spleen cells of the immunized Balb/c mice were fused in vitro. Five monoclonal antibodie (McAb) strains were obtained by HIT and sub-cloned and respectely named A1,A2,A4,A5 and F4. The immunoglobulin subtypes of the five McAbs were identified as IgG1,Ig2b,Ig2b,Ig2b and IgG1. All of the HI titers of ascites were ovre 13 log2. The five McAbs has not across reaction with AIV H7,AIV H9,IBV,NDV and EDS-76 V.Double antibodies sandwich ELISA technique for detecting AIV H5 was established with McAb,chicken-anti-AIV polyclonal antibodies and HRP labeled PcAb. The results showed that the ELISA has high sensitivity and could detect AIV H5 and not react to the AIV H7, AIV H9, NDV, IBV, and EDS-76V. This method has a wonderful application future in detection of AIV H5.45 lots of H5 subtype Avan influenza inactivated vaccine were detected by the ELISA established. we got the value of the optical density of HA antige. The results showed that this method could identify the subtype of the H5 subtype Avan influenza inactivated vaccine. The correlation was also compared between the the value of the optical density of HA antige and the HI antibody titers of these 45 lots of vaccines. We found there was low correlation. So this method was not enough to evaluate the efficacy of the inactivated vaccine now .