Development and evaluation of an enzyme-linked immunosorbent assay kit for determination of benzo(a)pyrene in water

Objective To develop an indirect competitive enzyme-linked immunosorbent assay kit for the determination of benzo(a)pyrene(BaP).Methods The optimum concentration of antigen and monoclonal antibody were determined with chessboard titration.The optimum conditions,including pH,saltion and methanol concentration,were separately measured with indirect competitive enzyme-linked immunosorbent assay(icELISA).Results The optimum coating concentration of antigen was 10 μg/mL,while the optimum dilution ratio of antibody was 5 000.The established icELISA for the detection of BaP exhibited a linear range of 5 to 50 ng/mL,a sensitivity of 3.30 ng/mL,and a half inhibitory concentration(IC₅₀)of 13.74 ng/mL in the conditions of pH 7.4,0.01 mol/L of saltion concentration and 10%of methanol.The average recovery was 105%in water samples.Conclusion These results indicate that the icELISA kit developed could be applied in rapid screening detection of BaP in water samples.