title: Competitive immunoassay for analysis of bisphenol A in children's sera using a specific antibody.
abstract: Bisphenol A (BPA) has been reported as a potential estrogenic substance that could affect human health and reproduction. In this study, a monoclonal antibody (Mab) against BPA was produced after the immunization of Balb/c mice with a conjugate of 4,4-bis (4-hydroxyphenyl) valeric acid coupling with keyhole limpet hemocyanin (BVA-KLH). The obtained Mab showed higher affinity against BPA and lower cross-reactivity toward 4,4'-sulfonyldiphenol, diphenolic acid, hydroquinone, salicylic acid, and other common phenolic compounds. Basing on the Mab, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed. Under the optimal conditions, the limit of detection (LOD) was found to be 0.1 ng mL(-1) with the linear working range of 0.45-10.56 ng mL(-1). After sample extraction, the fortified serum samples were detected with intra- and inter-assay recovery ranges of 81.2-92.9 and 84.4-94.4 %, respectively. Then, 100 children's sera were screened by ic-ELISA. The result showed that 54 % of the serum samples were BPA-positive. The positive samples were purified by immuno-affinity column (IAC) and further confirmed by high-performance liquid chromatography (HPLC) with fluorescence detector measured at λ ex/λ em 228/310 nm in acetonitrile-water solution (v:v, 40:60). The analysis of the unknown samples showed that ic-ELISA agreed well with the HPLC results. It also revealed that the ELISA developed here could be a useful tool for screening BPA in children's sera before the validation of HPLC.
authors.affiliation: School of Life Science, Jiangxi Science and Technology Normal University, Nanchang, Jiangxi Province, 330013, China.
authors.forename: Fanfan
authors.initials: F
authors.lastname: Yang
authors.affiliation: School of Life Science, Jiangxi Science and Technology Normal University, Nanchang, Jiangxi Province, 330013, China.
authors.forename: Long
authors.initials: L
authors.lastname: Xu
authors.affiliation: School of Life Science, Jiangxi Science and Technology Normal University, Nanchang, Jiangxi Province, 330013, China.
authors.forename: Lixin
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Nanchang Center for Disease Control and Prevention, Nanchang, Jiangxi, Province, 330006, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Life Science, Jiangxi Science and Technology Normal University, Nanchang, Jiangxi Province, 330013, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Meng
authors.affiliation: School of Life Science, Jiangxi Science and Technology Normal University, Nanchang, Jiangxi Province, 330013, China. lilirenrong@hotmail.com.
authors.forename: Renrong
authors.initials: R
authors.lastname: Liu
mesh_terms: D000368:Aged; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001559:Benzhydryl Compounds / Q000097:blood / Q000276:immunology*; D002648:Child; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004967:Estrogens / Q000097:blood / Q000276:immunology; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008807:Mice, Inbred BALB C; D010636:Phenols / Q000097:blood / Q000276:immunology*; D000098826:Bisphenol A Compounds
chemical_list: D000911:Antibodies, Monoclonal; D001559:Benzhydryl Compounds; D004967:Estrogens; D010636:Phenols; C006780:bisphenol A; D000098826:Bisphenol A Compounds
keywords: Bisphenol A; ELISA; HPLC; Immuno-affinity column; Monoclonal antibody; Sera