Kangyao Zhang

CdS:Mn quantum dot-functionalized g-C 3 N 4 nanohybrids as signal-generation tags for photoelectrochemical immunoassay of prostate specific antigen coupling DNAzyme concatamer with enzymatic biocatalytic precipitation

A new photoelectrochemical (PEC) immunosensor based on Mn-doped CdS quantum dots (CdS:Mn QDs) on g-C3N4 nanosheets was developed for the sensitive detection of prostate specific antibody (PSA) in biological fluids. The signal derived from the as-synthesized Cd:Mn QDs-functionalized g-C3N4 nanohybrids via a hydrothermal method and was amplified through DNAzyme concatamers on gold nanoparticles accompanying enzymatic biocatalytic precipitation. Experimental results by UV-vis absorption spectra and photoluminescence revealed that CdS:Mn QDs/g-C3N4 nanohybrids exhibited higher photocurrent than those of CdS:Mn QDs and g-C3N4 alone. Upon addition of target PSA, a sandwich-type immunoreaction was carried out between capture antibodies and the labeled detection antibodies. Accompanying introduction of gold nanoparticles, the labeled initiator strands on the AuNPs triggered hybridization chain reaction and the formation of DNAzyme concatamers in the presence of hemin. The formed DNAzyme catalyzed 4-chloro-1-naphthol (4-CN) to produce an insoluble/insulating precipitate on the Mn:CdS QDs/g-C3N4, and blocked the light harvesting of Mn:CdS QDs/g-C3N4, thus resulting in the decreasing photocurrent. Under optimal conditions, the immunosensor exhibited good photocurrent responses for determination of target PSA, and allowed detection of PSA at a concentration as low as 3.8pgmL-1. The specificity, reproducibility and precision of this system were acceptable. Significantly, this methodology was further evaluated for analyzing human serum samples, giving well-matched results with referenced PSA enzyme-linked immunosorbent assay (ELISA) method.

Reduced graphene oxide/BiFeO3 nanohybrids-based signal-on photoelectrochemical sensing system for prostate-specific antigen detection coupling with magnetic microfluidic device

A novel magnetic controlled photoelectrochemical (PEC) sensing system was designed for sensitive detection of prostate-specific antigen (PSA) using reduced graphene oxide-functionalized BiFeO3 (rGO-BiFeO3) as the photoactive material and target-triggered hybridization chain reaction (HCR) for signal amplification. Remarkably enhanced PEC performance could be obtained by using rGO-BiFeO3 as the photoelectrode material due to its accelerated charge transfer and improved the visible light absorption. Additionally, efficient and simple operation could be achieved by introducing magnetic controlled flow-through device. The assay mainly involved in anchor DNA-conjugated magnetic bead (MB-aDNA), PSA aptamer/trigger DNA (Apt-tDNA) and two glucose oxidase-labeled hairpins (H1-GOx and H2-GOx). Upon addition of target PSA, the analyte initially reacted with the aptamer to release the trigger DNA, which partially hybridized with the anchor DNA on the MB. Thereafter, the unpaired trigger DNA on the MB opened the hairpin DNA structures in sequence and propagated a chain reaction of hybridization events between two alternating hairpins to form a long nicked double-helix with numerous GOx enzymes on it. Subsequently, the enzymatic product (H2O2) generated and consumed the photo-excited electrons from rGO-BiFeO3 under visible light irradiation to enhance the photocurrent. Under optimal conditions, the magnetic controlled PEC sensing system exhibited good photocurrent responses toward target PSA within the linear range of 0.001 - 100ng/mL with a detection limit of 0.31pg/mL. Moreover, favorable selectivity, good stability and satisfactory accuracy were obtained. The excellent analytical performance suggested that the rGO-BiFeO3-based PEC sensing platform could be a promising tool for sensitive, efficient and low cost detection of PSA in disease diagnostics.

CdTe/CdSe quantum dot-based fluorescent aptasensor with hemin/G-quadruplex DNzyme for sensitive detection of lysozyme using rolling circle amplification and strand hybridization.

Lysozyme with a small monomeric globular enzymatic protein is part of the innate immune system, and its deficiency can cause the increased incidence of disease. Herein, we devise a new signal-enhanced fluorescence aptasensing platform for quantitative screening of lysozyme by coupling with rolling circle amplification (RCA) and strand hybridization reaction, accompanying the assembly of CdTe/CdSe quantum dots (QDs) and hemin/G-quadruplex DNzyme. Initially, target-triggered release of the primer was carried out from DNA duplex via the reaction of the aptamer with the analyte, and the released primer could be then utilized as the template to produce numerous repeated oligonucleotide sequences by the RCA reaction. Following that, the formed long-stranded DNA simultaneously hybridized with the CdTe/CdSe QD-labeled probe and hemin/G-quadruplex DNzyme strand in the system, thereby resulting in the quenching of QD fluorescent signal through the proximity hemin/G-quadruplex DNzyme on the basis of transferring photoexcited conduction band electrons of quantum dots to Fe(III)/Fe(II)-protoporphyrin IX (hemin) complex. Under optimal conditions, the fluorescent signal decreased with the increasing target lysozyme within the dynamic range from 5.0 to 500nM with a detection limit (LOD) of 2.6nM at the 3sblank criterion. Intra-assay and interassay coefficients of variation (CVs) were below 8.5% and 11.5%, respectively. Finally, the system was applied to analyze spiked human serum samples, and the recoveries in all cases were 85-111.9%.

Plasmonic Enhancement Coupling with Defect-Engineered TiO2–x: A Mode for Sensitive Photoelectrochemical Biosensing

This work demonstrates that the photoelectric response of defect-engineered TiO2-x modified with Au nanoparticles can be modulated by oxygen vacancy concentration and excitation wavelength. When strongly plasmonic Au nanoparticles are anchored to defect-engineered TiO2-x by DNA hybridization, several times plasmonic enhancement of photocurrent occurs under 585 nm excitation, and it is employed as a novel signaling mode for developing an improved photoelectrochemical sensing platform. This signaling mode combined with exonuclease III-assisted target recycling amplification exhibits excellent analytical performance, which provides a novel photoelectrochemical detection protocol.

Bio-bar-code-based photoelectrochemical immunoassay for sensitive detection of prostate-specific antigen using rolling circle amplification and enzymatic biocatalytic precipitation

Methods based on photoelectrochemistry have been developed for immunoassay, but most involve in a low sensitivity and a relatively narrow detectable range. Herein a new bio-bar-code-based split-type photoelectrochemical (PEC) immunoassay was designed for sensitive detection of prostate-specific antigen (PSA), coupling rolling circle amplification (RCA) with enzymatic biocatalytic precipitation. The bio-bar-code-based immunoreaction was carried out on monoclonal anti-PSA antibody (mAb1)-coated microplate using primer DNA and polyclonal anti-PSA antibody-conjugated gold nanoparticle (pDNA-AuNP-pAb2) with a sandwich-type assay format. Accompanying the immunocomplex, the labeled primer DNA on gold nanoparticle readily triggered RCA reaction in the presence of padlock probe/dNTPs/ligase/polymerase. The RCA product with a long single-stranded DNA could cause the formation of numerous hemin/G-quadruplex-based DNAzyme concatamers. With the assistance of nicking endonuclease, DNAzyme concatamers were dissociated from gold nanoparticle, which catalyzed the precipitation of 4-chloro-1-naphthol in the presence of H2O2 onto CdS nanorods-coated electrode (as the photoanode for the generated holes). The formed insoluble precipitate inhibited the electron transfer from the solution to CdS nanorods-modified electrode by using ascorbic acid as the electron donor. Under the optimum conditions, the photocurrent of the modified electrode decreased with the increasing of PSA concentration. A detectable concentration for target PSA with this system could be achieved as low as 1.8pgmL-1. In addition, our strategy also showed good reproducibility, high specificity and accuracy matched well with commercial PSA ELISA kits for real sample analysis. These remarkable properties revealed that the developed PEC immunoassay has great potential as a useful tool for the detection of PSA in practical application.

Novel photoelectrochemical immunosensor for disease-related protein assisted by hemin/G-quadruplex-based DNAzyme on gold nanoparticles to enhance cathodic photocurrent on p-CuBi2O4 semiconductor

A novel p-type semiconductor material (p-CuBi2O4) is designed for the construction of split-type photoelectrochemical (PEC) immunosensor for alpha-fetoprotein (AFP) with the hemin assistant to enhance the cathodic photocurrent. Initially, the photocathode of PEC immunosensor is fabricated by p-CuBi2O4 on a layer of gold nanoparticles (AuNPs, as a front contact of p-CuBi2O4) to enhance the efficiency of charge separation. In the presence of target AFP, a sandwich-type immunoreaction was carried out in capture antibody-coated microplate by using detection antibody and hemin-based G-quadruplex (labeled on the AuNP) as the signal probe. Upon exonuclease I (Exo I) introduction, the enzyme digested the hemin/G-quadruplex-based DNAzyme to release the hemin[Fe(III)], which captured the generated electrons of p-CuBi2O4-based photocathode to enhance photocurrent via the reduction of hemin[Fe(III)] to hemin[Fe(II)] in PEC detection system. Under the optimal conditions, the split-type photocathodic immunosensor showed a wide linear dynamic range from 50pgmL-1 to 20ngmL-1 at a limit of detection (LOD) of 14.7pgmL-1 toward target AFP. Moreover, the PEC immunosensor also displayed high specificity and good reproducibility. Favorably, method accuracy was evaluated to analyze human serum specimens, and gave matched-well results in comparison with commercially available enzyme-linked immunosorbent assay (ELISA) method.

Optical transformation of a CdTe quantum dot-based paper sensor for a visual fluorescence immunoassay induced by dissolved silver ions

This work designs a simple low-cost visual fluorescence immunoassay for disease-related biomarkers (carcinoembryonic antigen, CEA, used as a model protein) in biological fluids, based on the structural and optical transformation of CdTe quantum dots (QDs) immobilized on paper induced by dissolved silver ions (Ag+) from silver nanoparticles (AgNPs) via a cation-exchange reaction. A sandwich-type immunoreaction was initially carried out in a removable polystyrene high-binding microplate coated with monoclonal anti-CEA capture antibody by using AgNP-labeled polyclonal anti-CEA antibody as the detection antibody. Thereafter, the carried AgNPs accompanying the sandwiched immunocomplexes were dissolved by acid to release numerous silver ions, which induced the ion-exchange reaction with the immobilized CdTe QDs on the paper (attached onto the microplate lid) for the Cd-to-Ag transformation, thus resulting in the quenching of the visual fluorescence from the CdTe QDs owing to Ag2Te formation. Under optimal conditions, the fluorescence intensity decreased with the increasing CEA concentration from 0.02 to 50 ng mL-1 with a detection limit of 5.6 pg mL-1. Further, a visual assay based on a CdTe QD-based paper sensor was developed for CEA detection, and 5.0 pg mL-1 CEA could be discriminated with the naked eye. In addition, our strategy displayed high specificity, good reproducibility and acceptable accuracy for analyzing human serum specimens with consistent results obtained using the commercialized enzyme-linked immunosorbent assay (ELISA) method.

Reduced graphene oxide-functionalized FeOOH for signal-on photoelectrochemical sensing of prostate-specific antigen with bioresponsive controlled release system

A new and signal-on photoelectrochemical (PEC) sensing platform was successfully designed for the sensitive detection of prostate-specific antigen (PSA), using reduced graphene oxide- functionalized iron oxyhydroxide (FeOOH-rGO) as the photoactive material, accompanying target-responsive controlled release system to achieve the signal amplification. Introduction of rGO as electron mediator greatly facilitated the electron transfer from FeOOH to electrode under visible light, which inhibited the electron-hole recombination to enhance the photo-activity of FeOOH-rGO. Additionally, the bioresponsive release system was controlled via the reaction of target PSA with the aptamer capped glucose-loading mesoporous silica nanoparticle (MSN) to release numerous glucose molecules (as the electron donors) for the amplification of the photocurrent generated from FeOOH-rGO. Thus, more glucose molecules could be released and enhanced photocurrents could be obtained with the increasing PSA concentrations. Experimental results showed that the photocurrents of the PEC sensing platform were linearly dependent on the logarithm of PSA concentrations from 1.0pg/mL to 100ng/mL. Moreover, the PEC sensing system afforded good stability and specificity, and its accuracy matched well with the commercial PSA enzyme-linked immunosorbent assay (ELISA) kit. The excellent performance of the PEC sensing platform indicated its promising prospect as a useful tool for PSA detection in practical application.

Cu2+-Doped SnO2 Nanograin/Polypyrrole Nanospheres with Synergic Enhanced Properties for Ultrasensitive Room-Temperature H2S Gas Sensing

The organic-inorganic nanohybrids are emerging as one of the most attractive sensing materials in the area of gas sensors and usually exhibit some advanced properties because of synergetic/complementary effects between organic molecules and inorganic components. This work demonstrates a novel class of organic-inorganic nanohybrids, Cu2+-doped SnO2 nanograin/poly pyrrole nanospheres, for the sensitive room-temperature H2S gas sensing. Doping Cu2+ in SnO2 nanograins remarkably enhances the surface potential barrier by tailoring surface defects. After polymerizing pyrrole surrounded nanograins in aqueous media to form the organic-inorganic nanohybrids, the resulting nanoheterojunctions further improve the sensitivity. Additionally, the nanohybrids-based sensor provides high surface area and abounding reaction sites to accelerate gas diffusion and adsorption as well as the electron transfer. Compare with pristine SnO2 nanograins alone, the sensitivity of using the nanohybrids increases 7 times for the detection of 50-ppm of H2S. The response and recovery rate can increase 27 and 22 times at room temperature, respectively. Significantly, this work provides an attractive material for the real-time monitoring of H2S, whereas the insights into organic-inorganic composite interactions within the sensing mechanism may pave the way for designing functional materials with tailored properties.

Carbon Dots/g-C3N4 Nanoheterostructures-Based Signal-Generation Tags for Photoelectrochemical Immunoassay of Cancer Biomarkers Coupling with Copper Nanoclusters

A class of 0-dimensional/2-dimensional (0D/2D) nanoheterostructures based on carbon quantum dots (CQDs) and graphitic carbon nitride (g-C3N4) was designed as the signal-generation tags for the sensitive photoelectrochemical (PEC) immunoassay of prostate-specific antigen (PSA) coupling with the copper nanoclusters (CuNCs). Combination of CQDs with g-C3N4 promoted the photoexcited electron/hole separation and largely increased the photocurrents of the nanoheterostructures. Initially, a sandwich-type immunoreaction was carried out on monoclonal anti-PSA antibody-coated microplate by using PSA aptamer linked with CuNCs as the tracer. Accompanying the immunocomplex, the carried CuNCs were dissolved under acidic conditions. The as-released copper ions from the CuNCs could be captured onto the CQDs/g-C3N4 nanoheterostructures via the amino-group on the CQD surface as well as the -NHx (x = 1, 2, 3) of g-C3N4 nanosheets. The strong coordination of the Lewis basic sites on the CQDs/g-C3N4 with Cu2+ decreased the photocurrent of the nanoheterostructures. Under optimal conditions, CQDs/g-C3N4 nanoheterostructures displayed good photocurrent responses for the detection of PSA within the dynamic linear range of 0.02-100 ng mL-1 and a limit of detection (LOD) of 5.0 pg mL-1. This method was also evaluated for quantitative screening of human PSA serum specimens by using the referenced electrochemiluminescent enzyme-linked immunoassay (ECL-ELIA) and gave good matched results between two methods. Additionally, this system was beneficial to explore the charge-separation and photoinduced electron transfer mechanism in the photoelectrochemical sensing protocols.