A fluorescence polarization immunoassay method for detection of the bisphenol A residue in environmental water samples based on a monoclonal antibody and 4′-(aminomethyl)fluorescein

Abstract

Based on a sensitive monoclonal antibody against bisphenol A (BPA) and a new tracer named BVA-AMF, a homogeneous fluorescence polarization immunoassay (FPIA) was developed and applied in the determination of bisphenol A in environmental water samples. BVA was selected as the hapten to couple with bovine serum albumin and the conjugate was used as the immunogen for the monoclonal antibody production. Three fluorescein-labeled BVA tracers with different structures (BVA-AMF, BVA-EDF, and BVA-lysFITC) were synthesized. Under the same optimal conditions, BVA-AMF showed the highest sensitivity for FPIA and the detection of BPA exhibited a limit of detection of 5.60 ng mL À1 , an IC 50 of 140 ng mL À1 and a dynamic range of 11.32-904.21 ng mL À1 approximately. In this assay, the monoclonal antibody against BPA showed high specificity towards BPA with less than 0.15% cross-reactivity against several other similar compounds. Four different kinds of water samples were analyzed, with recoveries being 87.91-114.28%. The detection standard curve of BPA exhibited a good linearity (R 2 ¼ 0.9913, n ¼ 3).Compared with ELISA and HPLC methods, FPIA showed reliability and a high correlation with ELISA of 0.9964 and HPLC of 0.9971. The proposed immunoassay technique is suitable for detection of BPA in authentic environmental water samples.

Fluorescence polarization assay for the simultaneous determination of bisphenol A, bisphenol F and their diglycidyl ethers in canned tuna

“…In general, the optimal tracer concentration was determined by its fluorescence intensity at least ten times higher than the background signal from working buffer. [27] Thus, 8 nM of Dex-fl was selected as the final working concentration in this study. With a fixed concentration of tracers, the working concentration of mPPARα-LBD* was determined according to the K d of 7.04 nM.…”

Evaluation and Optimization of Three Different Immunoassays for Rapid Detection Zearalenone in Fodders

“…Small molecules in solution have a smaller fluorescence polarization value because of a fast spin, and the macro molecule vice versa (Lippolis et al 2014). Thus, in FPIA, the fraction of bound and unbound tracers can be calculated without having to conduct a physical separation of them (Huang et al 2015;Udenfriend 2014). FPIA has been commercially used for the detection of some small molecular, although, in some cases, they give only qualitative determinations.…”

Donor/acceptor nanoparticle pair-based singlet oxygen channeling homogenous chemiluminescence immunoassay for quantitative determination of bisphenol A

“…This makes homogeneous immunoassay practical for the rapid analysis of food and environmental samples. For example, a FPIA homogeneous immunoassay has been reported for the determination of BPA [28], but its sensitivity was limited, and its single tube design restricted rapid detection and highthroughput. There is much demand for a better homogenous immunoassay for the rapid, sensitive, and high-throughput analysis of BPA in environmental samples.…”

“…Enzyme-linked immunosorbent assay (ELISA) [25], fluorescence immunoassay (FI) [26], chemiluminescence enzymelinked immunosorbent assay (CL-ELISA) [27], and fluorescence polarization immunoassay (FPIA) [28] have been developed to detect BPA in environmental and food samples. ELISA, FI, and CL-ELISA are heterogeneous methods.…”