Abstract:

The method for quantitative analysis of AFB1 was established by using the microsphere array technology platform. Monoclonal anti-aflatoxin B1 was used in this study. Through optimizing the coupling antigen and determining critical saturated concentration of monoclonal anti-aflatoxin B1 as well as optimizing incubation time of antigen and antibody, the quantitative detection of AFB1 was established eventually. The results showed that, the yield of the coupled microballoon sphere was between 70.08%-80.80%, the best dosage of the coupling antigen was 100 μg. The median fluorescence intensity (MFI) detected by the same dosage of the coupling antigen and the identical concentration of the monoclonal anti-aflatoxin B1, increased along with the extending foster time and became stable finally. When the foster time was 24 h and 48 h, the MFI was the biggest with 2.0 ng/μL concentration of the monoclonal anti-aflatoxin B1. As referring to the international current standard of IC50, the standard curve showed that the detection sensitivity was 2.33 ng/mL (0.1165ng), the detection linear equation was y=0.0006x+0.0014, the cross reactivity rate of the monoclonal anti-aflatoxin B1 with AFB2, AFG1 and ZEN were 9.34%, 2.88% and<0.10%. The quantitative detection of AFB1 was established successfully in this study. The examination limitation of the standard curve was conformed to the China’s limitation standard of AFB1 in the food and feed. This method was suitable for the massive sample examination.