Preparation And Characterization Of The Monoclonal Antibodies Against Influenza A (H1N1) Virus

ObjectivePreparing specialized anti-influenza A (H1N1) Virus monoclonal antibodies, and charactering biological function of the antibodies.Method6~8 weeks female BALB/c mice were immunized with DNA vaccine which express hemagglutinin H1 (A/California/04/2009). Mosuse spleen B cells were fused with plasmocytoma cells NS1. Hybridoma cells were screened by indirect ELISA, then did the subcloning to obtain the cell line which specialized and steady secreting anti-influenza A (H1N1) Virus monoclonal antibody.ResultObtained 4 monoclonal antibody cell line which can steady secreting anti-influenza A(H1N1) virus hemagglutinin H1 subtype, named as 41H2, 68E5, 41E7and26D11. Among these antibodies, 68E5, 41E7 and 26D11were charactered as IgG1; 41H2 was charactered as IgG2a, all the antibodies'light chain was kappa. Lenti-pseudovirus test shows 26D11 could recogenize the constructive sites, had great ablity to neutralizing H1N1(A/California/04/2009), was neutralizing antibody. 41H2 and 41E7 were proved to be recogenizing the linear epitope by western blotting test. Our experiment shows 26D11, 41H2 and 41E7 could only specialized reacting to influenza A virus H1 subtype, and there was no crosslink reaction to the other 15 HA subtype.ConclusionSpecialized anti-influenza A (H1N1) Virus monoclonal antibodies were generated. And combined the polyclonal antibody, we also developed new ELISA kit to 2009 H1N1(Swine Flu) HA antigen.