Establishment of Time-resolved Fluorescence Immunoassay for Progesterone Residues in Animal-derived Foods
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Abstract:
In this study, progesterone was taken as the research object, a new antigen was designed and prepared, and specific rabbit antibody was obtained through animal immunization. Based on the obtained antigen and antibody, key parameters, such as the concentration of antigen and antibody, the type of reaction buffer and its pH value, were further optimized. Finally, a time-resolved fluorescence Immunoassay (TRFIA) for rapid detection of progesterone residues in animal-derived foods was established for the first time in China. In this study, the optimal experimental conditions were determined as follows: the concentration of antigen was 0.30 μg/mL, the concentration of europium labeled antibody was 0.40 μg/mL, the pH of TBST working buffer was 7.2, and the standard curve was established. The linear range of the method was 0.46~6.94 μg/L, the IC50 reached 1.79 μg/L, the cross-reaction rate with testosterone was 1.25%, and there was no cross-reaction with estradiol, estriol and other hormones. The average spiking recovery rate of the actual samples was 82.0%~113.0%, and the coefficient of variation within and between batches was less than 5%. The correlation with the analysis results obtained by high-performance liquid chromatography was good (r2 = 0.988). The results indicate that the TRFIA detection method established in this study was accurate, reliable, specific, and sensitive, and can be used for the rapid detection of progesterone residues in animal-derived foods.
