Development Of The Polyclonal Antibodies Against MON And Application For The Determination Of Mon Residues

This paper includes three parts of research, synthesis of the antigen and development of the polyclonal antibodies against MON and application for the determination of MON residues in eggs, the results was shown as follows:1. Development of the antigen against MONIn production Monensin is existed as Monensin sodium salt. However, Monensin sodium salt has not any group which are directly coupled with protein conjugations such as BSA,OVA. So it can not synthesize immugen directly and must be modified to the substance contained of carboxyl to produce antibody. We applied succinate acid anhydride method for transformation to monensin-succinate acid anhydride derivative. The materials of the transformation were appraised by thin layer chromatography. The result of appraising is approved successfully.The Mon-HS was coupled with protein (BSA, OVA) by the mixed acid anhydride reaction method. The conjugation included the immunogen(Mon-HS-BSA) and the coating antigen(Mon-HS-OVA). The synthesized antigens were appraised by SDS-PAGE. The protein-hapten conjugate rate was 14.7:1(Mon-HS-BSA). 2. Development of the polyclonal antibodies against MONHigh titer polyclonal antibodies against Monensin were produced from rabbits immunized with the synthesized immunogen conjugates Mon-HS-BSA. Through the indirect competitive enzyme-linked immunosorbent assay(icELISA).The final titer of antibodies were 1:25600(No.2). Calibration graphs were prepared by plotting log (MON concentration) against percentage binding. The percentage binding was calculated from the absorbance obtained in the absence (Bo) and the presence (B) of MON in standards and samples as follows: binding= B/Bo. The I₅₀ of this assay was established to be 32.4ng /ml, and the sensitivity of detection was 2.75ng/ml. To test the speciality of antibodies, SAL and HAN and SMM were used in the competitive ELISA as the competitive antigen. The result proved that the antibodies shows no cross-eaction to the other three antibiotics.