Development of hybridoma cell lines excreting monoclonal antibodies against zearalenone

Two hybridoma cell lines excreting monoclonal antibodies against Zearalenone (ZEN), coded Z2A4 and Z8D6, respectively, were obtained by fusing murine Sp2/0 cells with spleen cells from 8～10-week-old BalB/c mice immunized with ZEN-BSA conjugate and subcloning for 4 to 5 cycles .The ascites containing monoclonal antibodies against ZEN was gained via inoculation of the hybridoma cells into the abdominal cavity of BalB/c mice. The monoclonal antibodies produced by the hybridoma cells were tested for subtypes and designated as IgG 1 for ZEN. The titer of antibodies in ascites was 1∶ 1.6×106 for Z2A4 and 1∶3.2×106 for Z8D6, the MW was both 150 040(150 kD) and the working concentration was 1∶40 000 and 1∶100 000. IgG concentration in purified ascites yielded by hybridoma cells of Z2A4 and Z8D6 reached 34 and 39 mg/ml respectively, and had high specificity to ZEN because there was no cross reaction between the monoclonal antibodies against ZEN and other mycotoxins. Affinity constants of Z2A4 and Z8D6 were 5.52×108 and 4.68×109 mol/L respectively.