Study On Preparation Of Polyclonal Antibody Against Penicillin And Rapid Detection Of Penicillin Residue

In this study, penicillin was coupled to BSA and OVA, four kinds of conjugates were obtained, PEN-EDC-BSA and PEN-BSA were immunizing antigen, PEN-EDC-OVA and PEN-OVA were envelope antigen. The result of SDS-PAGE indicated that penicillin were coupled to carrier proteins successfully, the efficiency of the coupling reaction of PEN-EDC-BSA,PEN-EDC-OVA,PEN-BSA,PEN-OVA were 12:1,16:1,6:1,7:1, respectively, which were calculated through determining the OD280 and OD260 of carrier proteins,penicillin and conjugates. Two groups(E group and S group) of rabbits were immunized by PEN-EDC-BSA and PEN-BSA, respectively, and the antiserums were preparated, then their titre were determined by indirect ELISA, the titres of E group were between 1:1600-1:12800, and S group were between 1:102400-1:204900. The IgG was purified by caprylic acid-ammonium sulphate method, the concentrations of IgG solution were between 18.15-27.42mg/ml, and their titre were not changed on the whole, then we chose S3,S4 as the best IgG solutions, their concentrations and titres were 23.97,24.27mg/ml and 1:102400,1:204900, respectively.Different concentration of IgG solation was mixed with different concentration SPA stabilizing buffer for preparing SPA diagnosticum. Then SPA coagglutination test which detecting penicillin was done, we got the conclusions that the optimal SPA concentration was 5%, the optimum ratio of SPA stabilizing buffer and IgG was 2:1. The minimum penicillin detecting ability of SPA-COA to detect penicillin was 10μg/L.