Establishment And Preliminary Application Of Hybridomas Secreting Monoclonal Antibodies Against Clostridium Perfringens Enterotoxin

Hybridomas secreting monoclonal antibodies (McAbs)specific for Clostridium perfringens type A enterotoxin(CPE) were produced by fusion of Nso myelomas cells with spleen cells from BALB/C mice immunized with purified CPE .Wells containing hybridomas secreting immunoglobulin G (IgG) antibodies against CPE were specifically identified by an indirect enzyme-linked immunosorbent assay(ELISA),and two strains of ELISA-positive hybridomas were selected and cloned forth by limited dilution. All two hybridomas produced McAbs containing immunoglobulin G1 heavy chains. These hybridomas were then grown as ascitic tumors in mice,and McAbs were purified from the ascites fluids with DEAE.The specificity of the McAbs was demonstrated by RPLA and ELISA. The hybridomas with 96.8 average number of chromosome stablely secreted McAbs after subcultured twenty-one times. The optimal coating concentration of McAb was 2. 5μg/ml,working concentration of rabbit-anti-CPE positive serum was 1:400.The sensitivity of the McAb-AC-ELISA was 0.92ng/ml. Establishment of a double decker sandwhich-ELISA provided a simple,fast,sensitive and universally used method for the diagnosis of CPE.