Development of an Indirect Competitive ELISA Kit for the Rapid Detection of Benzopyrene Residues

This paper presents the generation of monoclonal antibodies (mAbs) with high specificity against benzopyrene (BaP) through cell fusion procedures, and the development of mAb-based indirect competitive enzyme-linked immunoabsorbent kit (icELISA kit) to detect BaP using one of these hybridomas (clone 2E12). The calibration curve of BaP-Kit with standard BaP inhibitor was typical sigmoid curve fitted to the four parameters logistic equation with the linear regression equation y = −28.662x + 132.87, the half-maximal inhibition concentration (IC50) of 0.779 μg/L and the sensitivity of 0.054 μg/L. The recovery of BaP spiked in beef was between 81.3 and 93.6 %. The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 6 %. Except for minor cross-reactivity with benzo[b]fluoranthene (2.7 %), other interference to the assay was negligible (<0.05 %). The validity of BaP kit in 4 °C was above 6 months. The sensitivity, specificity, simplicity, and reliability of BaP kit were confirmed by the procedure utilizing gas chromatography coupled to mass spectrometric (GC-MS) for the identification and quantification of BaP in barbecue. Their coincidence rate was 100 % compared with GC-MS. The kit was proved to be used for the rapid determination of BaP residues.