Preparation and Identification of Monoclonal Antibodies against Chicken Immunoglobulin G

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[Objective] To prepare monoclonal antibodies against chicken immunoglobulin G (IgG) and improve the diagnostic level of specific antibodies in chickens. [Method] Chicken IgG was isolated by saturated ammonium sulfate and purified by Sephadex G-200 column chromatography. Then the BALB/c mice were immunized by the chicken IgG, and the spleen cells were fused with mouse myeloma cells SP2/0. Finally, the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay (ELISA). [Result] Four hybridoma cell strains secreting monoclonal antibodies against chicken IgG were obtained and named as C44, C45, C67 and C68, and their ascites titers in indirect ELISA were 1∶640 000, 1∶320 000, 1∶640 000 and 1∶80 000, respectively. The monoclonal antibodies secreted by C44 and C45 could recognize light chains of chicken IgG and those secreted by C67 and C68 could recognize heavy chains of chicken IgG. They all could not recognize IgG from duck, rabbit and swine. Additionally, the Ig type identification results showed that they all belonged to IgG1. [Conclusion] Four cell strains of obtained hybridoma can stably produce the monoclonal antibodies against chicken IgG. [Objective] To prepare monoclonal antibodies against chicken immunoglobulin G (IgG) and improve the diagnostic level of specific antibodies in chickens. [Method] Chicken IgG was isolated by saturated ammonium sulfate and purified by Sephadex G-200 column chromatography. Then the BALB / c mice were immunized by the chicken IgG, and the spleen cells were fused with mouse myeloma cells SP2 / 0. Finally, the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay (ELISA). [Result] Four hybridoma cells secreting monoclonal antibodies against chicken IgG were obtained and named as C44, C45, C67 and C68, and their ascites titers in indirect ELISA were 1: 640,000, 1: 320,000, 1: 640,000 and 1: 80,000, respectively. The monoclonal antibodies secreted by C44 and C45 could recognize light chains of chicken IgG and those secreted by C67 and C68 could recognize heavy chains of chicken IgG. They all could not recognize IgG from duck, rabbit and swine. Additionally, the Ig type identification results showed that they all belonged to IgG1. [Conclusion] Four cell strains of obtained hybridoma can stably produce the monoclonal antibodies against chicken IgG.
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