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Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a fluorescent label
- Fan, Ziyan, Keum, Young Soo, Li, Qing X., Shelver, Weilin L., Guo, Liang-Hong
- Journal of environmental monitoring 2012 v.14 no.5 pp. 1345
- DNA, antibodies, antigens, benzopyrene, cross reaction, detection limit, equipment performance, estradiol, ethers, fluorescent dyes, fluorescent labeling, glass, immunoassays, microarray technology, pollutants, proteins, river water, screening, water pollution
- Indirect competitive immunoassays were developed on protein microarrays for the sensitive and simultaneous detection of multiple environmental chemicals in one sample. In this assay, a DNA/SYTOX Orange conjugate was employed as an antibody label to increase the fluorescence signal and sensitivity of the immunoassays. Epoxy-modified glass slides were selected as the substrate for the production of 4 × 4 coating antigen microarrays. With this signal-enhancing system, competition curves for 17β-estradiol (E2), benzo[a]pyrene (BaP) and 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) were obtained individually on the protein microarray. The IC₅₀ and calculated limit of detection (LOD) are 0.32 μg L⁻¹ and 0.022 μg L⁻¹ for E2, 37.2 μg L⁻¹ and 24.5 μg L⁻¹ for BaP, and 31.6 μg L⁻¹ and 2.8 μg L⁻¹ for BDE-47, respectively. LOD of E2 is 14-fold lower than the value reported in a previous study using Cy3 labeled antibody (Du et al., Clin. Chem, 2005, 51, 368–375). The results of the microarray immunoassay were within 15% of chromatographic analysis for all three pollutants in spiked river water samples, thus verifying the immunoassay. Simultaneous detection of E2, BaP and BDE-47 in one sample was demonstrated. There was no cross-reaction in the immunoassay between these three environmental chemicals. These results suggest that microarray-based immunoassays with DNA/dye conjugate labels are useful tools for the rapid, sensitive, and high throughput screening of multiple environmental contaminants.