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In contrast to the conventional means that the activity of horseradish peroxidase (HRP) is initiated and terminated by the additives of peroxides and strongly acidic stop solutions, this study demonstrates that the enzymatic activity of HRP is switched through the visible light irradiated graphene oxide (GO). And this visible light driven activity of HRP can realize time-precise control without the aids of peroxides (typically H2O2) and acidic stop solutions. The superoxide anions (O2•−) and photogenerated holes (h+) produced by the photo irradiated GO are responsible for activating HRP and the subsequent oxidation of the typical substrates, i.e., 3, 3′, 5, 5′-tetramethylbenzidine (TMB) and 2, 2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). It is also validated that the photoswitchable HRP-GO mixture can act as an efficient signal reporter of bioassays by taking the sandwich immunoassay of alpha-fetoprotein (AFP) as an example. The AFP can be detected sensitively and selectively in the linear range from 0.2 fg/mL to 1.0 ng/mL, with a very low detection limit of 0.1 fg/mL. Advantages of the photoswitchable HRP-GO mixture include high catalytic ability, precise time control, and free of additionally harmful reagents.

We propose herein an immobilization-free, split-mode cathodic photoelectrochemical (PEC) strategy coupled with a cascaded amplification for versatile biosensing. Taking DNA and microRNA (miRNA) as the model targets, the hybridization between the targets and the hairpin probe triggers the digestion of the probe DNA by T7 exonuclease (T7 Exo), thus to generate G-quadruplex (G4) forming sequences, and then the released targets (DNA or miRNA) initiate the subsequent cycling processes and generate a large amount of G4 forming sequences. Subsequently, the formed G4 sequences associate with hemin to form the G4/hemin DNAzyme, which catalytically produces 1,4-bezoquinone (BQ) for conjugating onto the surface of the chitosan (CS) deposited BiOI/ITO photocathode via the quinone-chitosan conjugation chemistry (QCCC). Under photo excitation, the covalently attached quinones can act as electron acceptors of bismuth oxyiodine (BiOI), promoting the photocurrent generation and thus allowing the elegant and “signal-on” mode for probing targets of interest. Highly sensitive and selective PEC bioassays are readily realized, with the detection limits down to 2.2 fM (for DNA) and 0.2 fM (for miRNA). Since no labeling and no electrode modification processes are needed, this split-mode PEC biosensing strategy is amenable to convenient, time/labor saving, and high-throughput detections. More significantly, it provides a novel concept to design immobilization-free and label-free cathodic PEC biosensing systems, and showcases promise in general and versatile bioanalysis research.

Photocathode based on p-type PbS quantum dots (QDs) combing a novel signal amplification strategy utilizing catalase (CAT) mimetics was designed and utilized for sensitive photoelectrochemical (PEC) detection of DNA. The bio-bar-coded Pt nanoparticles (NPs)/G-quadruplex/hemin exhibited high CAT-like activity following the Michaelis–Menten model for decomposing H2O2 to water and oxygen, whose activity even slightly exceeded that of natural CAT. The bio-bar-code as a catalytic label was conjugated onto the surface of PbS QDs modified electrodes through the formed sandwich-type structure due to DNA hybridization. Oxygen in situ generated by the CAT mimetics of the bio-bar-code of Pt NPs/G-quadruplex/hemin acted as an efficient electron acceptor of illuminated PbS QDs, promoting charge separation and enhancing cathodic photocurrent. Under optimal conditions, the developed PEC biosensor for target DNA exhibited a dynamic range of 0.2 pmol/L to 1.0 nmol/L with a low detection limit of 0.08 pmol/L. The high sensitivity of the method was resulted from the sensitive response of PbS QDs to oxygen and the highly efficient CAT-like catalytic activity of the bio-bar-coded Pt NPs/G-quadruplex/hemin.

An ultrasensitive photoelectrochemical (PEC) immunoassay based on signal amplification by enzyme mimetics was fabricated for the detection of mouse IgG (as a model protein). The PEC immunosensor was constructed by a layer-by-layer assembly of poly (diallyldimethylammonium chloride) (PDDA), CdS quantum dots (QDs), primary antibody (Ab1, polyclonal goat antimouse IgG), and the antigen (Ag, mouse IgG) on an indium-tin oxide (ITO) electrode. Then, the secondary antibody (Ab2, polyclonal goat antimouse IgG) combined to a bio-bar-coded Pt nanoparticle(NP)-G-quadruplex/hemin probe was used for signal amplification. The bio-bar-coded Pt NP-G-quadruplex/hemin probe could catalyze the oxidation of hydroquinone (HQ) using H2O2 as an oxidant, demonstrating its intrinsic enzyme-like activity. High sensitivity for the target Ag was achieved by using the bio-bar-coded probe as signal amplifier due to its high catalytic activity, a competitive nonproductive absorption of hemin and the steric hindrance caused by the polymeric oxidation products of HQ. For most important, the oxidation product of HQ acted as an efficient electron acceptor of the illuminated CdS QDs. The target Ag could be detected from 0.01pg/mL to 1.0ng/mL with a low detection limit of 6.0fg/mL. The as-obtained immunosensor exhibited high sensitivity, good stability and acceptable reproducibility. This method might be attractive for clinical and biomedical applications.

Due to the intrinsic hole oxidation reaction occurred on the photoanode surface, currently developed photoelectrochemical biosensors suffer from the interference from coexisting reductive species (acting as electron donor) and a novel design strategy of photoelectrode for photoelectrochemical detection is urgently required. In this paper, a self-operating photocathode based on CdS quantum dots sensitized three-dimensional (3D) nanoporous NiO was designed and created, which showed highly selective and reversible response to dissolved oxygen (acting as electron acceptor) in the electrolyte solution. Using glucose oxidase (GOD) as a biocatalyst, a novel photoelectrochemical sensor for glucose was developed. The commonly encountered interferents such as H2O2, ascorbic acid (AA), cysteine (Cys), dopamine (DA), etc., almost had no effect for the cathodic photocurrent of the 3D NiO/CdS electrode, though these substances were proved to greatly influence the photocurrent of photoanodes, which indicated greatly improved selectivity of the method. The method was applied to detect glucose in real samples including serum and glucose injections with satisfactory results. This study could provide a new train of thought on designing of self-operating photocathode in photoelectrochemical sensing, promoting the application of semiconductor nanomaterials in photoelectrochemistry.

The method provides an innovative dual functional sensors for mercury (II) ions and hydrogen peroxide. The addition of H(2)O(2) to the mixture of silver nanoparticles (AgNPs) and Hg(2+) induced color changes of the solution within several seconds even at 2.0 nM Hg(2+). Other metallic ions could not induce color change even at 10 μM. Of importance, this probe was not only successfully applied to detect Hg(2+), but also it could be used to sense H(2)O(2) at a concentration as low as 50 nM (by naked-eye). The outstanding sensitivity and selectivity property for Hg(2+) and H(2)O(2) resulted from the AgNPs mediated reduction of Hg(2+) to elementary Hg in the presence of H(2)O(2), causing the aggregation and colorimetric response of AgNPs. This sensitive and selective colorimetric assay opens up a fresh insight of development facile and fast detection methods for metal ions and biomolecules using the special catalytic reactivity of AgNPs.