Study On The Preparation And Characterization Of Monoclonal Antibodies Against Excretory-Secretory Antigen Of Muscle Larvae Of Trichinella Spiralis

This study aimed to establish B cell hybridoma strain secreting monoclonal antibodies (McAb) specific to excretory-secretory (ES) antigen of muscle larvae of Trichinella spiralis applying to hybridizable technique and the McAbs were characterized.BALB/C mice were immunized by ES antigen of muscle larvae of Trichinella spiralis. Their spleen lymph cell were fused with SP2/0 myeloma cell of mice followed to select hybridoma cell strain secreting the highest titer McAbs and extract and purify ascites. The activity of hybridoma cell strain culture medium and ascites were tested with Indirect ELISA. Indirect sandwich ELISA was performed to determine relative affinity. Competition ELISA was done to determine additivity effect and class or subclass of immunoglobulin was examined by agarose double immuodiffusion test. Cross reaction of McAbs were tested with adult worm antigen and egg antigen of Schistosoma japonicum, tachyzoites antigen of Toxoplasma gondii RH strain, cyst liquid antigen of Cysticercus cellulosae. cystic fluid antigen of Echinococcus granulosus, surface antigen, somatic antigen and ES antigen of adult worm of Trichinella spiralis, soluble antigen, 48h ES antigen, 72h ES antigen of muscle larvae. Electronicmicroscope was used to observe ultrastructure of hybridoma cell. The number of chromosome was counted. The stability of hybridoma cells secreting McAbs was evaluated.Two strains hybridoma cells secreting McAbs of anti-ES antigen of muscle larvae were acquired. B cell hybridoma characterizing rumour cell and plasma cell and including secreting granule of McAb was seen through electronic microscope. The mean number of chromosome was 98.6. Two strains hybridoma cells can stably secrete McAb during two mouths. Two strains McAbs belong to IgM. The titer of B2C12 strain culture medium and ascites were 1: 1280, 1: 2.048 X 104 respectively and that of El 1F11 were 1: 640, 1.024 X 104 respectively. The former had much higher relative affinity than the latter. Two McAbs can recognize the different ES antigenic determinant. The McAbs had no cross reaction with all the mentioned antigens.The produced hybridoma cell strains can secrete high titer and specificity McAbs. The characterized results demonstrated that McAbs were potential serodiagnosis reagents to establish diagnosis kit for detecting circulating antigen (CAg) and perfect material for producing gene engineering antibodies.