An Improved Chemiluminescence Immunoassay for the Ultrasensitive Detection of Aflatoxin B 1
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An Improved Chemiluminescence Immunoassay for the Ultrasensitive Detection of Aflatoxin B1 Junfeng Li 1 & Xiangyi Fang 1 & Yucong Yang 2 & Xiaoli Cheng 2 & Peng Tang 1
Received: 11 November 2015 / Accepted: 28 March 2016 # Springer Science+Business Media New York 2016
Abstract An indirect competitive chemiluminescence (CL) immunoassay based on Fe3O4@SiO2@Au magnetic nanoparticles (Au-SCMPs) has been optimized and developed for the detection of aflatoxin B1 (AFB1). To improve the detection sensitivity and efficiency, this method combines 2′,6′dimethylcarbonylphenyl-10-sulfopropyl acridinium-9carboxylate 4′-NHS ester (NSP-DMAE-NHS) as a new kind of high efficient luminescence reagent with a simplified separation procedure based on the optimized Au-SCMPs. Superparamagnetic nanoparticles were coated with different sizes of Au colloids (18, 30, and 50 nm), and their capacities in immobilization of bovine serum albumin (BSA) were investigated. The BSA-AFB 1 conjugate (BSA-AFB 1 ) was immobilized on the optimized Au-SCMPs and competed with the free AFB1 for specially binding the NSP-DAME-NHSlabeled anti-AFB1 antibody (mAb-AFB1). After the indirect competitive immunoreactions and magnetic separation, the CL of the nanoparticles was measured by a homemade luminescent measurement system. Under the optimum conditions, the IC50 value was 0.07 ng mL−1 and the limit of detection (LOD) of AFB1 was 0.01 ng mL−1, respectively. The result shows a much lower IC50 and LOD than that typically achieved Electronic supplementary material The online version of this article (doi:10.1007/s12161-016-0499-1) contains supplementary material, which is available to authorized users. * Xiangyi Fang [email protected]
1
Physics Department, School of Science, Xi’an Jiaotong University, Xi’an, People’s Republic of China
2
Department of Clinical Laboratory, The First Affiliated Hospital of Medical College, Xi’an Jiaotong University, Xi’an, People’s Republic of China
by ELISA. This proposed immunoassay method was rapid, low-cost, and suitable for the detection of AFB1. Keywords Chemiluminescence immunoassays . Aflatoxin B1 . Acridinium ester . Fe3O4@SiO2@Au magnetic nanocomposites
Introduction In recent years, magnetic nanoparticles are attracting great interest due to their promising application in many areas such as drug delivery, food and food industry, magnetic resonance imaging, and environmental remediation (Blasco and Picó 2011; Edelman and Warach 1993; Pankhurst et al. 2003; Zhang et al. 2010). To overcome the problems of toxicity and agglomeration limiting the application of magnetic nanoparticles in biological field, magnetic nanoparticles were often coated with various kinds of shells (Deng et al. 2005). The silica shell is one of the most appropriate layers for its good chemical stability, excellent biocompatibility, and water dispersibility (Ahangaran et al. 2013; Hui et al. 2011). Also, silica shell facilitates further coupling interaction with some guest molecules because of the abundant amounts of functional groups which is a grea
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