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Enzymatic Oxydate-Triggered Self-Illuminated Photoelectrochemical Sensing Platform for Portable Immunoassay Using Digital Multimeter

Author:
Shu, Jian, Qiu, Zhenli, Zhou, Qian, Lin, Youxiu, Lu, Minghua, Tang, Dianping
Source:
Analytical chemistry 2016 v.88 no.5 pp. 2958-2966
ISSN:
1520-6882
Subject:
biomarkers, chemiluminescence, detection limit, electrical equipment, electronic circuits, enzyme-linked immunosorbent assay, glucose, glucose oxidase, graphene, hydrogen peroxide, instrumentation, nanogold, prostate-specific antigen, proteins
Abstract:
Herein a novel split-type photoelectrochemical (PEC) immunosensing platform was designed for sensitive detection of low-abundance biomarkers (prostate-specific antigen, PSA, used in this case) by coupling a peroxyoxalate chemiluminescence (PO-CL) self-illuminated system with digital multimeter (DMM) readout. The PEC detection device consisted of a capacitor/DMM-joined electronic circuit and a PO-CL-based self-illuminated cell. Initially, reduced graphene oxide-doped BiVO₄ (BiVO₄-rGO) photovoltaic materials with good photoelectric properties was integrated into the capacitor/DMM-joined circuit for photocurrent generation in the presence of hydrogen peroxide (H₂O₂, as the hole-trapping reagent). A sandwich-type immunoreaction with target PSA was carried out in capture antibody-coated microplates by using glucose oxidase/detection antibody-conjugating gold nanoparticle (pAb₂-AuNP-GOx). Accompanying the sandwiched immunocomplex, the labeled GOx could oxidize glucose to produce H₂O₂. The as-generated H₂O₂ could act as the coreaction reagent to trigger the chemiluminescence of the peroxyoxalate system and the PEC reaction of the BiVO₄-rGO. Meanwhile, the self-illuminated light could induce photovoltaic material (BiVO₄-rGO) to produce a voltage that was utilized to charge an external capacitor. With the switch closed, the capacitor could discharge through the DMM and provide an instantaneous current. Different from conventional PEC immunoassays, the as-generated photoelectron was stored in the capacitor and released instantaneously to amplify the photocurrent. Under the optimal conditions, the transient current increased with the increasing target PSA concentration in the dynamic working range from 10 pg mL–¹ to 80 ng mL–¹ with a detection limit (LOD) of 3 pg mL–¹. This work demonstrated for the first time that the peroxyoxalate CL system could be used as a suitable substitute of physical light source to apply in PEC immunoassay. In addition, this methodology afforded good reproducibility, precision, and high specificity, and the method accuracy matched well with the commercial PSA ELISA kit. Importantly, the developed split-type photoelectrochemical immunoassay could not only avoid the interfering of the biomolecules relative to the photovoltaic materials but also eliminate the need of an exciting light source and expensive instrumentation, thus representing a user-friendly and low-cost assay protocol for practical utilization in quantitative low-abundance proteins.
Agid:
5304544