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

Overview

Journal Biosensors (Basel)

Specialty Biotechnology

Date 2024 Dec 27

PMID 39727853

Authors

Peerapong Jeeno

Sumed Yadoung

Marninphan Thongkham

Pichamon Yana

Udomsap Jaitham

Sakaewan Ounjaijean

Zhen-Lin Xu

Korawan Sringarm

Surat Hongsibsong

Affiliations

Soon will be listed here.

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 IC 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.

References

Manousi N, Zachariadis G . Recent Advances in the Extraction of Polycyclic Aromatic Hydrocarbons from Environmental Samples. Molecules. 2020; 25(9). PMC: 7249015. DOI: 10.3390/molecules25092182. View

Matschulat D, Deng A, Niessner R, Knopp D . Development of a highly sensitive monoclonal antibody based ELISA for detection of benzo[a]pyrene in potable water. Analyst. 2005; 130(7):1078-86. DOI: 10.1039/b503636d. View

Bukowska B, Sicinska P . Influence of Benzo(a)pyrene on Different Epigenetic Processes. Int J Mol Sci. 2021; 22(24). PMC: 8707600. DOI: 10.3390/ijms222413453. View

Zhu Y, Ma Y, Lu M, Zhang Y, Li A, Liang X . Efficient Production of Human Norovirus-Specific IgY in Egg Yolks by Vaccination of Hens with a Recombinant Vesicular Stomatitis Virus Expressing VP1 Protein. Viruses. 2019; 11(5). PMC: 6563233. DOI: 10.3390/v11050444. View

Fismes J, Perrin-Ganier C, Empereur-Bissonnet P, Morel J . Soil-to-root transfer and translocation of polycyclic aromatic hydrocarbons by vegetables grown on industrial contaminated soils. J Environ Qual. 2002; 31(5):1649-56. DOI: 10.2134/jeq2002.1649. View

Gerde P, Hanberg A, Jernstrom B, Johansson C, Kyrklund T, Rannug A . Cancer risk assessment, indicators, and guidelines for polycyclic aromatic hydrocarbons in the ambient air. Environ Health Perspect. 2002; 110 Suppl 3:451-88. PMC: 1241197. DOI: 10.1289/ehp.110-1241197. View

Torres O, Alving C, Matyas G . Synthesis of Hapten-Protein Conjugate Vaccines with Reproducible Hapten Densities. Methods Mol Biol. 2016; 1403:695-710. DOI: 10.1007/978-1-4939-3387-7_39. View

Amro W, Al-Qaisi W, Al-Razem F . Production and purification of IgY antibodies from chicken egg yolk. J Genet Eng Biotechnol. 2019; 16(1):99-103. PMC: 6296578. DOI: 10.1016/j.jgeb.2017.10.003. View

Meng X, Li Y, Zhou Y, Zhang Y, Yang L, Qiao B . An enzyme-linked immunosorbent assay for detection of pyrene and related polycyclic aromatic hydrocarbons. Anal Biochem. 2014; 473:1-6. DOI: 10.1016/j.ab.2014.12.002. View

10.

Wu S, Li H, Yin X, Si Y, Qin L, Yang H . Preparation of Monoclonal Antibody against Pyrene and Benzo [a]pyrene and Development of Enzyme-Linked Immunosorbent Assay for Fish, Shrimp and Crab Samples. Foods. 2023; 11(20). PMC: 9602326. DOI: 10.3390/foods11203220. View

11.

Yadoung S, Ishimatsu R, Xu Z, Sringarm K, Pata S, Thongkham M . Development of IgY-Based Indirect Competitive ELISA for the Detection of Fluoroquinolone Residues in Chicken and Pork Samples. Antibiotics (Basel). 2022; 11(11). PMC: 9686788. DOI: 10.3390/antibiotics11111512. View

12.

Scharnweber T, Fisher M, Suchanek M, Knopp D, Niessner R . Monoclonal antibody to polycyclic aromatic hydrocarbons based on a new benzo[a]pyrene immunogen. Fresenius J Anal Chem. 2002; 371(5):578-85. DOI: 10.1007/s002160101012. View

13.

Liang Y, Liu X, Liu Y, Yu X, Fan M . Synthesis of three haptens for the class-specific immunoassay of O,O-dimethyl organophosphorus pesticides and effect of hapten heterology on immunoassay sensitivity. Anal Chim Acta. 2008; 615(2):174-83. DOI: 10.1016/j.aca.2008.03.050. View

14.

Jiang W, Zhang H, Li X, Liu X, Zhang S, Shi W . Monoclonal antibody production and the development of an indirect competitive enzyme-linked immunosorbent assay for screening spiramycin in milk. J Agric Food Chem. 2013; 61(46):10925-31. DOI: 10.1021/jf404027b. View

15.

. Some non-heterocyclic polycyclic aromatic hydrocarbons and some related exposures. IARC Monogr Eval Carcinog Risks Hum. 2010; 92:1-853. PMC: 4781319. View

16.

Felemban S, Vazquez P, Moore E . Future Trends for In Situ Monitoring of Polycyclic Aromatic Hydrocarbons in Water Sources: The Role of Immunosensing Techniques. Biosensors (Basel). 2019; 9(4). PMC: 6955691. DOI: 10.3390/bios9040142. View

17.

Larsson A, Karlsson-Parra A, Sjoquist J . Use of chicken antibodies in enzyme immunoassays to avoid interference by rheumatoid factors. Clin Chem. 1991; 37(3):411-4. View

18.

Zhao N, Wu W, Cui S, Li H, Feng Y, Guo L . Effects of Benzo[a]pyrene-DNA adducts, dietary vitamins, folate, and carotene intakes on preterm birth: a nested case-control study from the birth cohort in China. Environ Health. 2022; 21(1):48. PMC: 9074263. DOI: 10.1186/s12940-022-00859-7. View