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Issue 5, 2012
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Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a fluorescent label

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Abstract

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.

Graphical abstract: Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a fluorescent label

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Article information


Submitted
29 Nov 2011
Accepted
14 Feb 2012
First published
15 Feb 2012

J. Environ. Monit., 2012,14, 1345-1352
Article type
Paper

Sensitive immunoassay detection of multiple environmental chemicals on protein microarrays using DNA/dye conjugate as a fluorescent label

Z. Fan, Y. S. Keum, Q. X. Li, W. L. Shelver and L. Guo, J. Environ. Monit., 2012, 14, 1345
DOI: 10.1039/C2EM10956E

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