Ultrasensitive Fluorescence Immunoassay for Detection of Bisphenol A in Milk Products Using Functionalized Gold Nanopart
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Ultrasensitive Fluorescence Immunoassay for Detection of Bisphenol A in Milk Products Using Functionalized Gold Nanoparticles as Probe Wei Ji 1 & Lingyun Du 1 & Yuanfu Zhang 1 & Guofu Liu 2 & Shuhao Wang 1
Received: 27 November 2014 / Accepted: 18 March 2015 # Springer Science+Business Media New York 2015
Abstract An ultrasensitive fluorescence immunoassay was developed for detection of bisphenol A (BPA). Herein, thiolmodified single-strand DNA (SH-ssDNAs) and anti-BPA antibodies were simultaneously conjugated with gold nanoparticles (AuNPs), and the fluorescein isothiocyanate-labeled single-strand DNA was hybridized with SH-ssDNAs to form a dual-codified probe. In the presence of BPA, a competitive immunoreaction was conducted between BPA and BPA coating antigens immobilized on microplate for the antiBPA antibodies of dual-codified probe. Then, in order to avoid the fluorescent signals quenched by AuNPs, restriction enzyme BamH І was employed to cut the fluorescent labels from immunocomplex, resulting in an increase of fluorescence emission. Under optimized conditions, the linear range for detection of BPA is from 1.0×10−2 to 1.0×103 ng/L with a low limit of detection of 3.4×10−3 ng/L. This method is of good sensibility, stability, and specificity, and it can be used to detect BPA in actual milk products and its migration from packaging materials into milk.
Keywords Fluorescenceimmunoassay . Functionalizedgold nanoparticles . Restriction enzyme . Bisphenol A
* Shuhao Wang [email protected] 1
Shandong Provincial Key Laboratory of Chemical Energy Storage and Novel Cell Technology, College of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng 252059, China
2
College of Life Science, Liaocheng University, Liaocheng 252059, China
Introduction Bisphenol A (BPA, 2,2-bis (4-hydroxyphenyl) propane), an indispensable organic raw material of elasticizer, antioxidant, hot stabilizer, and coating waterproofing material is widely used in cosmetics, paper products, food packaging, and water bottles (Staples et al. 1998; Vandenberg et al. 2007; Liao and Kannan 2011). However, numerous reports revealed that BPA possess certain estrogenic properties, constant exposure of BPA can induce the proliferation of cells and cause a series of potential pathological alteration of reproductive system even at extremely low exposure levels (Howdeshell et al. 1999; Hu et al. 2002; Takayanagi et al. 2006). And, BPA cannot participate in the process of normal metabolism, so a low content of compounds may bring discomfort. BPA in the organism mainly origins is from the intake of food (Schecter et al. 2010). Even though BPA was hydrophobic, BPA still can readily migrate into the food in various ways such as sustained exposure of food to BPA package or high temperatures process with BPA container or contact liposoluble components with BPA (Errico et al. 2014; Munguía-López et al. 2005; Cao and Corriveau 2008; Nam et al. 2010). Marsha K detected that the preschool children’s urine samples contained BPA (Marsha et al. 2011).
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