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Shu J.,Institute of Nanomedicine and Nanobiosensing | Qiu Z.,Institute of Nanomedicine and Nanobiosensing | Zhuang J.,Institute of Nanomedicine and Nanobiosensing | Xu M.,Institute of Nanomedicine and Nanobiosensing | Tang D.,Institute of Nanomedicine and Nanobiosensing
ACS Applied Materials and Interfaces | Year: 2015

An ultrasensitive photoelectrochemical (PEC) immunoassay protocol for quantitative detection of low-abundant proteins at a low potential was designed by utilizing porphyrin-sensitized titanium dioxide (TiO2) nanostructures. Experimental results demonstrated that the water-soluble 5,10,15,20-tetra(4-sulfophenyl)-21H,23H-porphyrin (TSPP) could be bound onto titanium dioxide via the sulfonic group. TSPP-sensitized TiO2 nanostructures exhibited better photoelectrochemical responses and stability in comparison with TiO2 nanoparticles alone under continuous illumination. Using carcinoembryonic antigen (CEA) as a model analyte, a typical PEC immunosensor by using TSPP-TiO2 as the affinity support of anti-CEA capture antibody (Ab1) to facilitate the improvement of photocurrent response was developed. Bioconjugates of secondary antibody and glucose oxidase with gold nanoparticles (Ab2/GOx-AuNPs) was introduced by an antigen-antibody immunoreaction. AuNP acted as a powerful scaffold to bind with bioactive molecules, while GOx catalyzed glucose to in situ generate hydrogen peroxide (H2O2). The generated H2O2 as a sacrificial electron donor could be oxidized by the photogenerated holes to assist the signal amplification at a low potential under light excitation, thus eliminating interference from other species coexisting in the samples. Under optimal conditions, the PEC immunosensor showed a good linear relationship ranging from 0.02 to 40 ng mL-1 with a low detection limit of 6 pg mL-1 CEA. The precision, reproducibility, and specificity were acceptable. In addition, the method accuracy was also evaluated for quantitatively monitoring human serum samples, giving results matching with the referenced CEA ELISA kit. © 2015 American Chemical Society.

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