A label-free ratiometric immunoassay using bioinspired nanochannels and a smart modified electrode.

Zhe Qiao, Zilian Jiang, Qiufen Luo, Hongfang Zhang, Jianbin Zheng

Analytica chimica acta 2021 Jun 01

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Labeling with redox reporter is often required in developing electrochemical bioassay for most proteins or nucleic acid biomarkers. Herein, a label-free ratiometric immunosensing platform is firstly developed by integrating the antibody-conjugated nanochannels with a smart modified electrode. The electrode modifier is the composite of C60, tetraoctylammonium bromide (TOA+) and Prussian blue (PB). Cyclic voltammograms of the ultimate C60-TOA+/PB modified electrode exhibited two pairs of peaks at 0.15 V and -0.13 V, ascribing to the redox of PB and C60, respectively. With the addition of K3[Fe(CN)6] in the electrolyte solution, the peaks of PB decreased due to the adsorption of [Fe(CN)6]3- while the peaks of C60 increased because of the formation of the ternary complex (TC) C60-TOA+-[Fe(CN)6]3-. As a result, the peak current ratio IPB/ITC decreased gradually with the increment of the concentration of [Fe(CN)6]3-. For the nanochannels-based immunosensing platform, the steric hindrance of the bioconjugated nanochannels varied with the loading amount of the target CA125, and thus [Fe(CN)6]3- passing through the channels was quantitatively affected. And the higher CA125 level was, the less [Fe(CN)6]3- concentration was. And thus, the ratio IPB/ITC monitored at the C60-TOA+/PB modified electrode increased with the increase of the concentration of CA125. The ratiometric immunoassay featured a linear calibration range from 1.0 U mL-1 to 100 U mL-1 with a low detection limit of 0.86 U mL-1. In addition, the ratiometric immunosensing platform demonstrated good specificity and stability as well as acceptable accuracy in overcoming the effect of electrode passivation which was an inherent problem of electroanalysis. Copyright © 2021 Elsevier B.V. All rights reserved.

Citation

Zhe Qiao, Zilian Jiang, Qiufen Luo, Hongfang Zhang, Jianbin Zheng. A label-free ratiometric immunoassay using bioinspired nanochannels and a smart modified electrode. Analytica chimica acta. 2021 Jun 01;1162:338476