Pyrenecarboxaldehyde@Graphene Oxide Acted as a Highly Efficient ECL Emitter and Target-Triggered the Recyclable Cascade System as an Amplifier for Ultrasensitive APE1 Activity Detection.

Herein, pyrenecarboxaldehyde@graphene oxide (Pyc@GO) sheets with highly efficient electrochemiluminescence (ECL) as emitters were prepared by a noncovalent combination to develop a neoteric ECL biosensing platform for the ultrasensitive assessment of human apurinic/apyrimidinic endonuclease1 (APE1) activity. Impressively, the pyrenecarboxaldehyde (Pyc) molecules were able to form stable polar functional groups on the surface of GO sheets through the noncovalent π-π stacking mechanism to prevent intermolecular restacking and simultaneously generate Pyc@GO sheets. Compared with the tightly packed PAH nanocrystals, the Pyc@GO sheets significantly reduced internal filtering effects and diminished nonactivated emitters to enhance ECL intensity and achieve strong ECL emission. Furthermore, the APE1-activated initiators could trigger the recyclable cascade amplified system based on the synergistic cross-activation between catalytic hairpin assembly (CHA) and DNAzyme, which improved the signal amplification and transduction ability. Consequently, the developed ECL platform for the detection of APE1 activity displayed exceptional sensitivity with a low detection limit of 4.6 × 10-9 U·mL-1 ranging from 10-8 to 10-2 U·mL-1. Therefore, the proposed strategy holds great promise for the future development of sensitive and reliable biosensing platforms for the detection of various biomarkers.

Orderly Aggregated Catalytic Hairpin Assembly for Synchronous Ultrasensitive Detecting and High-Efficiency Co-Localization Imaging of Dual-miRNAs in Living Cells.

In this work, the orderly aggregated catalytic hairpin assembly (OA-CHA) was developed for synchronous ultrasensitive detection and high-efficiency colocalization imaging of dual-miRNAs by a carefully designed tetrahedral conjugated ladder DNA structure (TCLDS). Exactly, two diverse hairpin probes were fixed on tetrahedron conjugated DNA nanowires to form the TCLDS without fluorescence response, which triggered OA-CHA in the aid of output DNA 1 and output DNA 2 produced by targets miRNA-217 and miRNA-196a cycle to generate TCLDS with remarkable fluorescence response. Impressively, compared with the traditional CHA strategy, OA-CHA avoided the fluorescence group and quenching group from approaching again because of the spatial confinement effect to significantly enhance the fluorescence signal, resulting in the simultaneous ultrasensitive detection of dual-miRNAs with detection limits of 21 and 32 fM for miRNA-217 and miRNA-196a, respectively. Meanwhile, the TCLDS with lower diffusivity could achieve accurate localization imaging for reflecting the spatial distribution of dual-miRNAs in living cells. The strategy based on OA-CHA provided a flexible and programmable nucleic amplification method for the synchronous ultrasensitive detection and precise imaging of multiple biomarkers and had potential in disease diagnostics..

Ag@Pyc Nanocapsules as Electrochemiluminescence Emitters for an Ultrasensitive Assay of the APE1 Activity.

Herein, Ag@pyrenecarboxaldehyde nanocapsules (Ag@Pyc nanocapsules) as emitters were prepared to construct an ultrasensitive electrochemiluminescence (ECL) biosensor for the detection of the human apurinic/apyrimidinic endonuclease1 (APE1) activity. Ag nanoparticles on the surface of Pyc nanocapsules as coreaction accelerators could significantly promote coreactant peroxydisulfate (S2O82-) to generate massive reactive intermediates of sulfate radical anion (SO4•-), which interacted with the Pyc nanocapsules to achieve a strong ECL response. In addition, with the aid of APE1-triggered 3D DNA machine, trace target could be converted into a large number of mimic targets (MTs), which were positively correlated with the activity of APE1. Consequently, the proposed ECL biosensor realized an ultrasensitive detection of APE1 activity with an exceptional linear working range from 5 × 10-10 to 5 × 10-4 U·µL-1 and a lower limit of detection of 1.36 × 10-11 U·µL-1. This strategy provided a new approach to construct an efficient ternary system for the detection of biomolecules and early diagnosis of diseases.

Ultrasensitive Fluorescence Detection and Accurate Colocalization Visualization of Dual-miRNAs in Cancer Cells Based on the Conjugated Chain Reaction of Multifunctional Pentagon DNA Nanostructures.

Herein, a multifunctional pentagon DNA nanostructure (MPDN) was assembled by the hybridization of a circular DNA scaffold containing five different fragments with five diverse DNA oligonucleotides for simultaneous sensitive detection and accurate colocalization imaging of dual-miRNAs in cancer cells. Exactly, the MPDN could specifically and efficiently internalize into folate (FA) receptor-overexpressed cells via specific binding of FA and the FA receptor to distinguish cancer cells from normal cells and transform trace amounts of targets miRNA-21 and miRNA-155 into substantial FAM and Cy5-labeled DNA polymers as the signal probe to generate two remarkable fluorescence emissions, realizing simultaneously sensitive detection of dual-miRNAs. Impressively, compared with traditional small fragment DNA probes with high fluidity, the DNA copolymers with extremely low diffusivity kept it in the originally generated position to achieve the colocalization imaging of dual-miRNAs more accurately for revealing the spatial expression information of dual-miRNAs in tissues and cells. This strategy provided programmable tool to simultaneously detect and accurately colocate dual-miRNAs for understanding normal physiology and the tumor mechanism.

A photoelectrochemical biosensor based on methylene blue sensitized Bi5O7I for sensitive detection of PSA.

Herein, bismuth oxyiodide (Bi5O7I) was used as a signal probe to construct an effective sensitization structure with methylene blue (MB), combined with protein conversion strategy, and a photoelectrochemical (PEC) biosensor was constructed for sensitive detection of prostate-specific antigen (PSA). The designed biosensor had a high sensitivity and a low detection limit (LOD) of 0.047 fg mL-1, which opened up a simple way for the detection of PSA and showed a good application prospect in clinical and medical fields.

A new photoelectrochemical biosensor based on FeOOH and exonuclease III-aided dual recycling signal amplification for HPV-16 detection.

Herein, based on iron oxyhydroxide (FeOOH) as the photoactive material and exonuclease III (exo III)-aided dual recycling signal amplification, a new photoelectrochemical (PEC) biosensor was successfully developed for human papillomavirus-16 (HPV-16) detection with a wide linear range from 0.5 fM to 1 nM and a low detection limit of 0.17 fM.

Ultrasensitive photoelectrochemical microRNA biosensor based on doxorubicin sensitized graphitic carbon nitride assisted by a target-activated enzyme-free DNA walker.

Herein, a photoelectrochemical biosensor was successfully constructed on the basis of a sensitization strategy of doxorubicin sensitized graphitic carbon nitride for the ultrasensitive detection of microRNA-141 with the assistance of a target-activated enzyme-free DNA walker.

Supersensitive Photoelectrochemical Aptasensor Based on Br,N-Codoped TiO2 Sensitized by Quantum Dots.

Here, we fabricated a novel photoelectrochemical (PEC) aptasensor based on Br,N-codoped TiO2/CdS quantum dots (QDs) sensitization structure with excellent energy level arrangement for supersensitive detection of carcinoembryonic antigen (CEA). The prepared Br,N-codoped TiO2 could reduce the energy bandwidth of TiO2 from 3.2 to 2.88 eV, which could dramatically reduce the basic signal and obviously broaden the absorption of light (400-700 nm). In addition, the energy bandwidth of Br,N-codoped TiO2 (2.88 eV) matched well with that of CdS QDs (2.4 eV), making CdS QDs an ideal signal enhancer for amplifying the photocurrent signal of Br,N-codoped TiO2. More importantly, the constructed Br,N-codoped TiO2/CdS QDs sensitization structure with narrow energy level gradient enabled the effective promotion of electron-transfer capability and dramatic improvement of photoelectric conversion efficiency. Simultaneously, a small amount of the CEA was transformed into substantial single-chain DNA (T-DNA) via exonuclease III (Exo-III)-assisted cycle strategy. Under optimum conditions, the designed PEC aptasensor demonstrated a wide detection range from 1 fg/mL to 1 ng/mL and a low detection limit as 0.46 fg/mL for CEA assay. This strategy prepared a new photoactive material to markedly improve photoelectric conversion efficiency and initiated a new way to realize the highly sensitive PEC biomolecules detection.

An ultrasensitive aptasensor based on self-enhanced Au nanoclusters as highly efficient electrochemiluminescence indicator and multi-site landing DNA walker as signal amplification.

Gold nanoclusters (Au NCs) have been shown to be prospective nanoscale electrochemiluminescence (ECL) materials that are being extensively explored in bioanalysis. However, the low ECL efficiency of Au NCs has been a bottleneck barrier for their better bioapplications. To overcome this disadvantage, a low oxidation potential co-reactant N,N-diisopropylethylenediamine (DPEA) was first used to prepare self-enhanced Au NCs (Au-DPEA NCs) for drastically enhancing the ECL efficiency of Au NCs in this study. In addition, an efficient multi-site landing DNA walker with multidirectional motion track and rapid payloads release compared to directional DNA walker was constructed for converting target mucin 1 (MUC1) to intermediate DNA and achieving significant signal amplification. On the basis of the Au-DPEA NCs as efficient ECL signal labels and multi-site landing DNA walker as signal amplification strategy, an ECL aptasensor was established for the ultrasensitive detection of MUC1 in the range from 1 fg mL-1 to 1 ng mL-1 with a limit of detection down to 0.54 fg mL-1. The results demonstrated that the present study opened a new research direction for the development of high-efficiency Au NCs indicator as well as ultrasensitive ECL sensing platform for applications in clinical and bioanalysis.

Cosensitization Strategy with Cascade Energy Level Arrangement for Ultrasensitive Photoelectrochemical Protein Detection.

Here, a photoelectrochemical (PEC) biosensor was established by a cosensitization strategy with cascade energy level arrangement for ultrasensitive detection of prostate-specific antigen (PSA). The proposed cosensitization strategy was based on the well-matched energy level arrangement of four kinds of organic photoactive materials, in which poly{4,8-bis[5-(2-ethylhexyl)thiophen-2-yl]benzo[1,2- b:4,5- b']dithiophene-2,6-diyl- alt-3-fluoro-2-[(2-ethylhexyl)-carbonyl]thieno[3,4- b]thiophene-4,6-diyl} (PTB7-Th) was used as the photoactive material and perylenetetracarboxyl diimide (PDI), fullerene (nano-C60), and polyaniline (PANI) were employed as the sensitizers. The resulting PTB7-Th/PDI/nano-C60/PANI cascade cosensitization structure with narrow energy level gradient (<0.54 eV) could effectively improve electron transfer capability, obviously raise light energy utilization and significantly enhance photoelectric conversion efficiency, leading to dramatically enhanced photocurrent response. Using PSA as a target model, the proposed PEC biosensor exhibited high sensitivity and excellent stability with a wide detection range from 1 fg/mL to 0.1 ng/mL and a detection limit of 0.43 fg/mL. Moreover, the proposed PEC biosensor provides a cascade cosensitization strategy that could significantly improve PEC performances and open up a promising platform to establish high selectivity, stability, and ultrasensitive analytical techniques.

An ultrasensitive electrochemiluminescence biosensor for the detection of concanavalin A based on Au nanoparticles-thiosemicarbazide functionalized PtNi nanocubes as signal enhancer.

A sandwich-configuration electrochemiluminescence (ECL) biosensor was constructed for detecting concanavalin A (ConA) based on peroxydisulfate/oxygen (S2O82-/O2) system. In this work, the gold nanoflower modified Zn-doped SnO2 was used as a substrate to adsorb recognition element horseradish peroxidase (HRP) for binding ConA. Then, Au nanoparticles-thiosemicarbazide functionalized PtNi nanocubes (AuNPs-TSC-PtNi NCs), as a novel ECL signal tracer, were incubated onto the electrode through a specific carbohydrate-ConA interaction, thus achieving a sandwiched structure. The integration of amplifying effect of both TSC and PtNi NCs on the ECL of S2O82-/O2 system endowed the biosensor a high sensitivity. The linear range for ConA detection was from 0.0010ng/mL to 10ng/mL with a detection limit of 0.0002ng/mL (S/N=3).

A signal-on electrochemiluminescence biosensor for detecting Con A using phenoxy dextran-graphite-like carbon nitride as signal probe.

A novel signal-on electrochemiluminescence (ECL) biosensor for detecting concanavalin A (Con A) was fabricated with phenoxy dextran-graphite-like carbon nitride (DexP-g-C3N4) as signal probe. In this construction strategy, the nanocomposites of three-dimensional graphene and gold nanoparticles (3D-GR-AuNPs) were used as matrix for high loading of glucose oxidase (GOx), which served as recognition element for bounding Con A. Con A further interacted with DexP-g-C3N4 through a specific carbohydrate-Con A interaction to achieve a sandwiched scheme. With the increase of Con A incubated onto the electrode, the ECL signal resulted from DexP-g-C3N4 would enhance, thus achieving a signal-on ECL biosensor for Con A detection. Due to the integration of the virtues of 3D-GR-AuNPs and the excellent ECL performance of DexP-g-C3N4, the prepared biosensor exhibits a wide linear response range from 0.05 ng/mL to 100 ng/mL and a low detection limit of 17 pg/mL (S/N=3).

Graphene-multiwall carbon nanotube-gold nanocluster composites modified electrode for the simultaneous determination of ascorbic acid, dopamine, and uric acid.

In this paper, graphene-multiwall carbon nanotube-gold nanocluster (GP-MWCNT-AuNC) composites were synthesized and used as modifier to fabricate a sensor for simultaneous detection of ascorbic acid (AA), dopamine (DA), and uric acid (UA). The electrochemical behavior of the sensor was investigated by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and differential pulse voltammetry (DPV) techniques. The combination of GP, MWCNTs, and AuNCs endowed the electrode with a large surface area, good catalytic activity, and high selectivity and sensitivity. The linear response range for simultaneous detection of AA, DA, and UA at the sensor were 120-1,701, 2-213, and 0.7-88.3 µM, correspondingly, and the detection limits were 40, 0.67, and 0.23 µM (S/N=3), respectively. The proposed method offers a promise for simple, rapid, selective, and cost-effective analysis of small biomolecules.

Overoxidized polyimidazole/graphene oxide copolymer modified electrode for the simultaneous determination of ascorbic acid, dopamine, uric acid, guanine and adenine.

In the present work, a novel strategy based on overoxidized polyimidazole (PImox) and graphene oxide (GO) copolymer modified electrode was proposed for the simultaneous determination of ascorbic acid (AA), dopamine (DA), uric acid (UA), guanine (G) and adenine (A). The copolymer was characterized by the scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier transform infrared (FT-IR), X-ray photoelectron spectroscopy (XPS) and electrochemical impedance spectroscopy (EIS). Due to the synergistic effects between PImox and GO, the proposed electrode exhibited excellent electrochemical catalytic activities and high selectivity and sensitivity toward the oxidation of AA, DA, UA, G and A. The peak separations between AA and DA, AA and UA, UA and G, and G and A were 140 mV, 200 mV, 380 mV and 300 mV, respectively. The linear response ranges for AA, DA, UA, G and A were 75-2275 µM, 12-278 µM, 3.6-249.6 µM, 3.3-103.3 µM and 9.6-215 µM, respectively, and corresponding detection limits were 18 µM, 0.63 µM, 0.59 µM, 0.48 µM and 1.28 µM.

Detection of organophosphorus pesticides using potentiometric enzymatic membrane biosensor based on methylcellulose immobilization.

In this research, a new potentiometric enzymatic membrane biosensor for the detection of organophosphorus pesticides (OPs) was constructed. The basic element of this biosensor was a pH electrode modified with an immobilized acetylcholinesterase layer formed by entrapment with methylcellulose, N,N-dimethylformamide, and bovine serum albumin. The response characteristics of the biosensor were measured and discussed. It was shown that there is a good linear relationship between the inhibition rate and the negative logarithm of OPs concentration in the range from 10(-7) to 10(-5) mol/L, with the detection limits of 10(-7) mol/L for the five pesticides. Moreover, the biosensor could resist the disturbances of below 10(-6) mol/L of Cu(2+) and Pb(2+), and below 10(-5) mol/L of Cd(2+). In addition, the measurement results obtained by the biosensor method were in good agreement with those obtained by the gas chromatography method. This method was successfully applied to detect OPs that remained in the potted lettuce.