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. 2014 Oct 13;14(10):18886-97.
doi: 10.3390/s141018886.

Heterogeneous electrochemical immunoassay of hippuric acid on the electrodeposited organic films

Young-Bong Choi  1 , Nam-Hyuk Kim  2 , Seung-Hoi Kim  3 , Gun-Sik Tae  4 , Hyug-Han Kim  5
Affiliations

Heterogeneous electrochemical immunoassay of hippuric acid on the electrodeposited organic films

Young-Bong Choi et al. Sensors (Basel). .

Abstract

By directly coordinating hippuric acid (HA) to the ferrate (Fe) as an electron transfer mediator, we synthesized a Fe-HA complex, which shows a good electrochemical signal and thus enables the electrochemical immunoanalysis for HA. We electrodeposited organic films containing imidazole groups on the electrode surface and then bonded Ni ion (positive charge) to induce immobilization of Fe-HA (negative charge) through the electrostatic interaction. The heterogeneous competitive immunoassay system relies on the interaction between immobilized Fe-HA antigen conjugate and free HA antigen to its antibody (anti-HA). The electric signal becomes weaker due to the hindered electron transfer reaction when a large-sized HA antibody is bound onto the Fe-HA. However, in the presence of HA, the electric signal increases because free HA competitively reacts with the HA antibody prior to actual reaction and thus prevents the HA antibody from interacting with Fe-HA at the electrode surface. This competition reaction enabled an electrochemical quantitative analysis of HA concentration with a detection limit of 0.5 μg mL(-1), and thus allowed us to develop a simple and rapid electrochemical immunosensor.

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Figures

Figure 1.
Figure 1.
Schematic representation of the fabricating SPCEs/Organic film/Ni/Fe-HA for heterogeneous electrochemical immunoassay.
Figure 2.
Figure 2.
Preparation of Fe-conjugated HA antigen [Fe(CN)5(amp-HA)]3−/2− (Fe-HA).
Figure 3.
Figure 3.
Cyclic voltammograms for (a) SPCEs/Organic film and (b) SPCEs/Organic film/Ni surface recorded in phosphate buffer solution containing 2 mM K3Fe(CN)6. Cyclic voltammetry was conducted at a scan rate of 100 mVs−1. Inset: Anode currents at 0.375 V versus Ag/AgCl as a function of the number of cycles, ranging between five and 30.
Figure 4.
Figure 4.
Cyclic voltammograms for: (a) SPCEs/Organic film and (b) SPCEs/Organic film/Ni surfaces, recorded in phosphate buffer solution containing 2 mM K3Fe(CN)6. Cyclic voltammetry was conducted at a scan rate of 100 mVs−1, and the voltammograms are compared to the bare SPCEs, dashed line.
Figure 5.
Figure 5.
Differential pulse voltammograms of the SPCEs/Organic film/Ni/Fe-HA with variable anti-HA (0 ∼ 5 mg mL−1). Inset shows the calibration curve of the cathodic DPV peak current of SPCEs/Organic film/Ni/Fe-HA at 0.45 V versus Ag/AgCl as a function of the anti-HA concentrations.
Figure 6.
Figure 6.
Differential pulse voltammograms of the SPCEs/Organic film/Ni/Fe-HA with variable HA (0 ∼ 5 mg mL−1). Inset shows the calibration curve of the anodic DPV peak current of SPCEs/Organic film/Ni/Fe-HA at 0.45 V versus Ag/AgCl as a function of the HA concentrations (N = 6, r = 0.9943).
Figure 7.
Figure 7.
Cyclic voltammogram of the SPCEs/Organic film/Ni/Fe-HA in 2 mM ascorbic acid solution at scan rate of 0.10 V s−1.
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