Yaqin Chai

Ultrasensitive electrochemical immunosensor for clinical immunoassay using thionine-doped magnetic gold nanospheres as labels and horseradish peroxidase as enhancer.

A new signal amplification strategy based on thionine (TH)-doped magnetic gold nanospheres as labels and horseradish peroxidase (HRP) as enhancer holds promise to improve the sensitivity and detection limit of the immunoassay for carcinoembryonic antigen (CEA), as a model protein. This immunoassay system was fabricated on a carbon fiber microelectrode (CFME) covered with a well-ordered anti-CEA/protein A/nanogold architecture. The reverse micelle method was initially used for the preparation of TH-doped magnetic gold nanospheres (nanospheres), and the synthesized nanospheres were then labeled on HRP-bound anti-CEA as a secondary antibody (bionanospheres). Sandwich-type protocol was successfully introduced to develop a new high-efficiency electrochemical immunoassay with the labeled bionanospheres toward the reduction of H2O2. Under optimized conditions, the linear range of the proposed immunoassay without HRP as enhancer was 1.2-125 ng/mL CEA, whereas the assay sensitivity by using HRP as enhancer could be further increased to 0.01 ng/mL with the linear range from 0.01 to 160 ng/mL CEA. The developed immunoassay method showed good precision, high sensitivity, acceptable stability and reproducibility, and could be used for the detection of real samples with consistent results in comparison with those obtained by the enzyme-linked immunosorbent assay (ELISA) method.

Electrochemical sensing of hydrogen peroxide using metal nanoparticles: a review

AbstractWe are reviewing the state of electrochemical sensing of H2O2 based on the use of metal nanoparticles. The article is divided into subsections on sensors based on nanoparticles made from Ag, Pt, Pd, Cu, bimetallic nanoparticles and other metals. Some sensors display high sensitivity, fast response, and good stability. The review is subdivided into sections on sensors based on heme proteins and on nonenzymatic sensors. We also discussed the challenges of nanoscaled sensors and their future aspects. FigureSensing mechanism of (A) mediator-based enzyme biosensor, (B) mediator-less enzyme biosensor and (C) nonenzymatic sensors with metal nanoparticles for the electrocatalytic reduction toward H2O2

In Situ Hybridization Chain Reaction Amplification for Universal and Highly Sensitive Electrochemiluminescent Detection of DNA

In this work, we describe a new universal and highly sensitive strategy for electrochemiluminescent (ECL) detection of sequence specific DNA at the femtomolar level via in situ hybridization chain reaction (HCR) signal amplification. The DNA capture probes are self-assembled on a gold electrode. The presence of the target DNA and two hairpin helper DNAs leads to the formation of extended dsDNA polymers through HCR on the electrode surface. The in situ, HCR-generated dsDNA polymers cause the intercalation of numerous ECL indicators (Ru(phen)(3)(2+)) into the dsDNA grooves, resulting in significantly amplified ECL signal output. The proposed strategy combines the amplification power of the DNA HCR and the inherent high sensitivity of the ECL technique and enables low femtomolar detection of sequence specific DNA. The developed strategy also shows high selectivity against single-base mismatch sequences, which makes our new universal and highly sensitive HCR-based method a useful addition to the amplified DNA detection arena.

Bienzyme functionalized three-layer composite magnetic nanoparticles for electrochemical immunosensors

The preparation, characterization and application of a three-layer magnetic nanoparticle composed of an Fe(3)O(4) magnetic core, a Prussian Blue (PB) interlayer and a gold shell (it can be abbreviated as Au-PB-Fe(3)O(4)) for an ultrasensitive and reproducible electrochemical immunosensing fabrication were described for the first time in this work. With the employment of the Au-PB-Fe(3)O(4) nanoparticle, a new signal amplification strategy was developed based on bienzyme (horseradish peroxidase and glucose oxidase) functionalized Au-PB-Fe(3)O(4) nanoparticles for an electrochemical immunosensing fabrication by using Carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP) as model systems, respectively. The experiment results show that the multilabeled Au-PB-Fe(3)O(4) nanoparticles exhibit satisfying redox electrochemical activity and high enzyme catalysis activity, which predetermine their utility in high sensitivity antibody detection schemes. Furthermore, this immunosensor could be regenerated by simply using an external magnetic field which ensured a reproducible immunosensor with high sensitivity.

Simultaneous electrochemical detection of multiple analytes based on dual signal amplification of single-walled carbon nanotubes and multi-labeled graphene sheets

In this work, a sandwich-type electrochemical aptasensor for simultaneous sensitive detection of platelet-derived growth factor (PDGF) and thrombin is fabricated. Reduced graphene oxide sheets (rGS) are used as matrices to immobilize the redox probes, which are subsequently coated with platinum nanoparticles (PtNPs) to form the PtNPs-redox probes-rGS nanocomposites. With the employment of the as prepared nanocomposites, a signal amplification strategy was described based on bienzyme (glucose oxidase and horseradish peroxidase) modified PtNPs-redox probes-rGS nanocomposites as the tracer labels for secondary aptamers (Apt II) through sandwiched assay. Gold nanoparticles functionalized single-walled carbon nanotubes (AuNPs@SWCNTs) as the biosensor platform enhance the surface area to capture a large amount of primary aptamers (Apt I), thus amplifying the detection response. The experiment results show that the multi-labeled PtNPs-redox probes-rGS nanocomposites display satisfying electrochemical redox activity and highly electrocatalytic activity of PtNPs and bienzyme, which exhibit high sensitivity for detection of proteins. The linear range of PDGF is 0.01-35 nM with a detection limit of 8 pM, while the linear ranges from 0.02 to 45 nM and a detection limit of 11 pM for thrombin are obtained.

An “Off–On” Electrochemiluminescent Biosensor Based on DNAzyme-Assisted Target Recycling and Rolling Circle Amplifications for Ultrasensitive Detection of microRNA

In this study, an off-on switching of a dual amplified electrochemiluminescence (ECL) biosensor based on Pb(2+)-induced DNAzyme-assisted target recycling and rolling circle amplification (RCA) was constructed for microRNA (miRNA) detection. First, the primer probe with assistant probe and miRNA formed Y junction which was cleaved with the addition of Pb(2+) to release miRNA. Subsequently, the released miRNA could initiate the next recycling process, leading to the generation of numerous intermediate DNA sequences (S2). Afterward, bare glassy carbon electrode (GCE) was immersed into HAuCl4 solution to electrodeposit a Au nanoparticle layer (depAu), followed by the assembly of a hairpin probe (HP). Then, dopamine (DA)-modified DNA sequence (S1) was employed to hybridize with HP, which switching off the sensing system. This is the first work that employs DA to quench luminol ECL signal, possessing the biosensor ultralow background signal. Afterward, S2 produced by the target recycling process was loaded onto the prepared electrode to displace S1 and served as an initiator for RCA. With rational design, numerous repeated DNA sequences coupling with hemin to form hemin/G-quadruplex were generated, which could exhibit strongly catalytic toward H2O2, thus amplified the ECL signal and switched the ON state of the sensing system. The liner range for miRNA detection was from 1.0 fM to 100 pM with a low detection limit down to 0.3 fM. Moreover, with the high sensitivity and specificity induced by the dual signal amplification, the proposed miRNA biosensor holds great potential for analysis of other interesting tumor markers.

Simultaneous voltammetric determination for DA, AA and NO2− based on graphene/poly-cyclodextrin/MWCNTs nanocomposite platform

In the present work, graphene sheets (GS) and multiwall carbon nanotubes (MWCNTs) were dispersed in the mixed solution of cyclodextrin (CD) and cyclodextrin prepolymer (pre-CD) and were used as modifier to fabricate chemical modified electrode to simultaneous detect dopamine (DA), ascorbic acid (AA) and nitrite (NO2(-)). CD cross-linked pre-CD (CDP) displays excellent film forming ability, which made the electrode stable. Comparing with CDP-GS, CDP-MWCNTs and CDP-GS-MWCNTs modified electrodes, the CDP-GS-MWCNTs displays higher catalytic activity and selectivity toward the oxidation of DA, AA and NO2(-), revealing that MWCNTs effectively inhibited the stacking of individual GS and enhanced the utilization of GS based composites. The host-guest chemical reaction ability of CD and π-π stacking interaction between detected molecules and GS-MWCNTs surface were considered as the main reasons of the successfully simultaneous detection of DA, AA and NO2(-). Cyclic voltammetry (CV), scanning electron microscopy (SEM) and different pulse voltammetry (DPV) were employed to characterize the biosensor. The linear response range for AA, DA and NO2(-) were 5 μM-0.48 mM, 0.15-21.65 μM and 5 μM-6.75 mM, respectively and the detection limits were 1.65 μM, 0.05 μM and 1.65 μM.

Simultaneous determination of ascorbic acid, dopamine, uric acid and tryptophan on gold nanoparticles/overoxidized-polyimidazole composite modified glassy carbon electrode

A novel electrode was developed through electrodepositing gold nanoparticles (GNPs) on overoxidized-polyimidazole (PImox) film modified glassy carbon electrode (GCE). The combination of GNPs and the PImox film endowed the GNPs/PImox/GCE with good biological compatibility, high selectivity and sensitivity and excellent electrochemical catalytic activities towards ascorbic acid (AA), dopamine (DA), uric acid (UA) and tryptophan (Trp). In the fourfold co-existence system, the peak separations between AA-DA, DA-UA and UA-Trp were large up to 186, 165 and 285 mV, respectively. The calibration curves for AA, DA and UA were obtained in the range of 210.0-1010.0 μM, 5.0-268.0 μM and 6.0-486.0 μM with detection limits (S/N=3) of 2.0 μM, 0.08 μM and 0.5 μM, respectively. Two linear calibrations for Trp were obtained over ranges of 3.0-34.0 μM and 84.0-464.0 μM with detection limit (S/N=3) of 0.7 μM. In addition, the modified electrode was applied to detect AA, DA, UA and Trp in samples using standard addition method with satisfactory results.

Immobilization of horseradish peroxidase on chitosan/silica sol-gel hybrid membranes for the preparation of hydrogen peroxide biosensor.

A simple and effective strategy for fabrication of hydrogen peroxide (H2O2) biosensor has been developed by entrapping horseradish peroxidase (HRP) in chitosan/silica sol-gel hybrid membranes (CSHMs) doped with potassium ferricyanide (K3Fe(CN)6) and gold nanoparticles (GNPs) on platinum electrode surface. The hybrid membranes are prepared by cross-linking chitosan (CS) with 3-aminopropyltriethoxysilane (APTES), while the presence of GNPs improved the conductivity of CSHMs, and the Fe(CN)6(3-/4-) was used as a mediator to transfer electrons between the electrode and HRP due to its excellent electrochemistry activity. UV-Vis absorption spectroscopy was employed to characterize the different components in the CSHMs and their interaction. The parameters influencing the performance of the resulting biosensor were optimized and the characteristic of the resulting biosensor was characterized by cyclic voltammetry and chronoamperometry. Linear calibration for hydrogen peroxide was obtained in the range of 3.5x10(-6) to 1.4x10(-3) M under the optimized conditions with the detection limit (S/N = 3) of 8.0x10(-7) M. The apparent Michaelis-Menten constant of the enzyme electrode was 0.93 mM. The enzyme electrode retained about 78% of its response sensitivity after 30 days. The system was applied for the determination of the samples, and the results obtained were satisfactory.

Au-nanoclusters incorporated 3-amino-5-mercapto-1,2,4-triazole film modified electrode for the simultaneous determination of ascorbic acid, dopamine, uric acid and nitrite.

A novel biosensor has been constructed by the electrodeposition of Au-nanoclusters (nano-Au) on poly(3-amino-5-mercapto-1,2,4-triazole) (p-TA) film modified glassy carbon electrode (GCE) and employed for the simultaneous determination of dopamine (DA), ascorbic acid (AA), uric acid (UA) and nitrite (NO(2)(-)). NH(2) and SH groups exposed to the p-TA layer are helpful for the electrodeposition of nano-Au. The combination of nano-Au and p-TA endow the biosensor with large surface area, good biological compatibility, electricity and stability, high selectivity and sensitivity and flexible and controllable electrodeposition process. In the fourfold co-existence system, the linear calibration plots for AA, DA, UA and NO(2)(-) were obtained over the range of 2.1-50.1 μM, 0.6-340.0 μM, 1.6-110.0 μM and 15.9-277.0 μM with detection limits of 1.1×10(-6) M, 5.0×10(-8) M, 8.0×10(-8) M and 8.9×10(-7) M, respectively. In addition, the modified biosensor was applied to the determination of AA, DA, UA and NO(2)(-) in urine and serum samples by using standard adding method with satisfactory results.