In-House Immunoglobulin Y-Based Immunoassay for Detecting Benzo[a]pyrene in Grilled Pork Samples

Abstract

Benzo[a]pyrene (B[a]P) is a hazardous polycyclic aromatic hydrocarbon that accumulates in several environmental matrices as a result of incomplete combustion. Its presence, carcinogenic properties, and tendency for bioaccumulation provide significant risks to human health and the environment. The objective of this study is to create an immunoassay for the detection of benzo[a]pyrene utilizing immunoglobulin Y antibodies. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was utilized to develop a speedy, straightforward, sensitive, and economical approach for detecting B[a]P residues. Following the immunization of hens with the hapten pyrenebutyric acid-bovine serum albumin (PyBA-BSA), the IgY antibody extracted from egg yolk was utilized to identify B[a]P residues. To evaluate antibody specificity, six PAH derivatives—PyBA, B[a]P, Chrysene, Benzo[b]fluoranthene, Benzo[a]anthracene, and Benzo[k]fluoranthene—were examined in the experiment to compete for binding with PyBA. The findings indicate that the antibody had considerable affinity for Chrysene (1.15%), Benzo[b]fluoranthene (311.32%), Benzo[k]fluoranthene (10.62%), Benzo[a]anthracene (22.82%), and PyBA (9.55%). Nonetheless, its affinity for B[a]P remained at 100%. The recovery range for grilled pork samples spiked with B[a]P doses of 10.00–0.1 μg/mL was 74.99% to 143.11%. This study utilized a polyclonal antibody, employing the IgY antibody for the inaugural development of an immunoassay to detect benzo[a]pyrene. The ELISA had a higher IC50 value compared to the other immunoassays; however, it yielded good results. This immunoassay signifies a substantial progression in environmental analytical chemistry, offering a cost-effective and accessible technique for the detection of B[a]P to protect human health and the environment.