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. 2015 Oct 20;7(10):4216-31.
doi: 10.3390/toxins7104216.

A Magnetic Nanoparticle Based Enzyme-Linked Immunosorbent Assay for Sensitive Quantification of Zearalenone in Cereal and Feed Samples

Xian Zhang  1 , Xin Wang  2 , Mengjiao Sun  3 , Xiaofeng Zhang  4 , Houhui Song  5 , Yaxian Yan  6 , Jianhe Sun  7 , Xiaoliang Li  8 , Weihuan Fang  9
Affiliations

A Magnetic Nanoparticle Based Enzyme-Linked Immunosorbent Assay for Sensitive Quantification of Zearalenone in Cereal and Feed Samples

Xian Zhang et al. Toxins (Basel). .

Abstract

A novel enzyme-linked immunosorbent assay based on magnetic nanoparticles and biotin/streptavidin-HRP (MNP-bsELISA) was developed for rapid and sensitive detection of zearalenone (ZEN). The detection signal was enhanced and the sensitivity of the assay was improved by combined use of antibody-conjugated magnetic nanoparticles and biotin-streptavidin system. Under the optimized conditions, the regression equation for quantification of ZEN was y = -0.4287x + 0.3132 (R² = 0.9904). The working range was 0.07-2.41 ng/mL. The detection limit was 0.04 ng/mL and IC50 was 0.37 ng/mL. The recovery rates of intra-assay and inter-assay ranged from 92.8%-111.9% and 91.7%-114.5%, respectively, in spiked corn samples. Coefficients of variation were less than 10% in both cases. Parallel analysis of cereal and feed samples showed good correlation between MNP-bsELISA and liquid chromatograph-tandem mass spectrometry (R² = 0.9283). We conclude that this method is suitable for rapid detection of zearalenone in cereal and feed samples in relevant laboratories.

Keywords: biotin-streptavidin; immunoassay; magnetic nanoparticles; quantification; zearalenone.

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Figures

Figure 1
Figure 1
Schematic diagrams of the preparation of the immunomagnetic nanoparticles cross-linked with anti-zearalenone monoclonal antibody (MNP-anti-ZEN) (A) and MNP-bsELISA (B).
Figure 2
Figure 2
Identification of conjugation of zearalenone with bovine serum albumin (ZEN-BSA) by indirect ELISA (A) and Western blotting (B). The molecular weight of BSA is about 66 KDa. Lane 1: ZEN-BSA, Lane 2: BSA.
Figure 3
Figure 3
Identification of the immunomagnetic nanoparticles cross-linked with anti-zearalenone monoclonal antibody (MNP-anti-ZEN) by indirect MNP-bsELISA.
Figure 4
Figure 4
Determination of suitable incubation time by indirect ELISA with (+ZEN) or without (−ZEN) addition of zearalenone.
Figure 5
Figure 5
Tri-parametric curve fitting of log concentration of zearalenone vs. inhibition index by MNP-bsELISA. The insert shows the standard curve for quantification of zearalenone.
Figure 6
Figure 6
Analysis of matrix interferences by different dilutions, with PBS, of corn extracts spiked with different concentrations of zearalenone by MNP-bsELISA.
Figure 7
Figure 7
Correlation of results obtained by both MNP-bsELISA and LC-MS/MS for zearalenone detection in natural cereal and feed samples.
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