Mercury ion (Hg2+) as a major environmental pollutant threatens human health even at very low concentrations, so it is essential to monitor mercury residues in food. In this study, Hg2+ was conjugated with protein carrier using 1-(4-Isothiocyanobenzyl) ethylenediamine N, N, N', N'-tetraacetic acid (ITCBE) as a bifunctional chelator. 7A1 monoclonal antibody (mAb) against Hg2+-ITCBE with high affinity (7.3 × 109 L/moL) and good specificity was obtained by cell fusion technology and performed to establish immunosensors. Immunochromatographic test strip using colloidal gold nanoparticles (AuNP with an average diameter of 18 nm) as signal reporter showed low sensitivity. Signal amplification probes including larger multi-branched gold nanoflowers (AuNF) and latex microspheres (LM) were employed to enhance the sensitivity of immunosensors. The visible limit of detection (vLOD) of the AuNF- and LM-based strip were determined to be 50 ng/mL and 25 ng/mL respectively, showing more sensitive than that of AuNP-based strip (200 ng/mL). Quantitative analysis showed that AuNF-based strip exhibited lower quantitative limit of detection (qLOD) (0.44 ng/mL) which was 20-fold lower than that of AuNP-based strip (8.92 ng/mL) for determination of Hg2+, and LM-based strip (0.49 ng/mL) was 18 times as sensitive as AuNP-based strip. In summary, the developed immunosensors using AuNF and LM as signal amplification probes exhibited excellent sensitivity and provided portable, on-site detection for Hg2+.
Keywords: Gold nanoflowers; Immunochromatographic test strip; Latex microspheres; Mercury (II) ion; Monoclonal antibody.
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