Change In Electron Transfer Resistance Research Articles

A label-free electrochemical impedimetric DNA biosensor for genetically modified soybean detection based on gold carbon dots.

A label-free electrochemical impedimetric biosensor was constructed based on gold carbon dots (GCDs) modified screen-printed carbon electrode for the detection of genetic modified (GM) soybean. The structure and property of GCDs were investigated. The GCDs can directly bind to single-stranded DNA probes through Au-thiol interaction and boost electric conductivity for the DNA sensor construction. The quantification of target DNA was monitored by the change of electron-transfer resistance (Ret) upon the DNA hybridization on sensor surface. Under the optimal conditions, the Ret response (vs. Ag reference electrode) increased with the logarithm of target DNA concentrations in a wide linear range of 1.0 × 10-7 - 1.0 × 10-13M with a detection limit of 3.1 × 10-14M (S/N = 3). It was also demonstrated that the proposed DNA sensor possessed high specificity for discriminating target DNA from mismatched sequences. Moreover, the developed biosensor was applied to detect SHZD32-1 in actual samples, and the results showed a good consistency with those obtained from the gel electrophoresis method. Compared with the previous reports for DNA detection, the label-free biosensor showed a comparatively simple platform due to elimination of complicated DNA labeling. Therefore, the proposed method showed great potential to be an alternative device for simple, sensitive, specific, and portable DNA sensor.

An amperometric biosensor for specific detection of glycated hemoglobin based on recombinant engineered fructosyl peptide oxidase

Here, we present a specific biosensor based on the detection of glycated hemoglobin (HbA1c) proteolytic digestion product, fructosyl valyl histidine (Fru-ValHis). A recombinant engineered fructosyl peptide oxidase (FPOX) enzyme with improved specificity was immobilized on the electrode surface modified by chitosan (CHIT), graphene oxide (GO) and gold nanoparticles (AuNPs). The biosensor exhibited a linear response toward different concentrations of Fru-ValHis ranging from 0.1 to 2 mM with a sensitivity of 8.45 µA mM-1 cm−2. Detection limit of the current biosensor for Fru-ValHis was 0.3 µM as the lowest quantity required giving a signal to a background. Analytical recovery of added Fru-ValHis in whole blood was 95.1–98.35% for FPOX/AuNPs/GO/CHIT/FTO electrode. For Fru-ValHis determination by FPOX-AuNPs-GO-CHIT/FTO electrode, within-run coefficient of variation (CV) was between 1.3% and 2.4% and between run CV was between 2.1% and 3.5%. A significant change in electron transfer resistance after the incubation of FPOX-modified electrode with Fru-ValHis was observed, while no response was achieved with control, indicating specific measurement of Fru-ValHis. Moreover, designed biosensor measured HbA1c in human blood samples and the results were well agreed with that obtained with NORUDIA™ N HbA1c diagnostic kit. Overall, suitable specificity of the engineered FPOX made the bioelectrode responded well to the Fru-ValHis level, which demonstrates a promising application for specific detection of HbA1c biomarker.

Fully printed one-step biosensing device using graphene/AuNPs composite

Driven by the growing need of simple, cost efficient and flexible sensing systems, we have designed here a fully printed Reduced Graphene Oxide (rGO) based impedimetric sensor for one step sensing of DNA. The DNA sensor was fabricated by stamping of layered rGO and rGO/gold nanoparticles/single stranded DNA (rGO/AuNPs/ssDNA) composites over PET substrates using wax-printing technique. rGO works as an excellent working electrode, while the AuNPs create a suitable environment for ssDNA immobilization. Counter and reference electrodes were previously screen-printed on the plastic substrate, making thus a compact and highly integrated sensing platform. The change in electron transfer resistance after hybridization with a target ssDNA specific of Coxsackie B3 virus was monitored using electrochemical impedance spectroscopy (EIS), finding a linear response in the range of concentrations 0.01–20 µM. The novel, simple and straightforward one-step printing process for fabrication of a biosensing device developed keeps in mind the growing need of large scale device manufacturing. The successful proof-of-concept for the detection of DNA hybridization can be extended to other affinity biosensors, taking advantage of the integration of the bioreceptor on the sensor surface. Such ready-to-use biosensor would lead to a one-step electrochemical detection.

Label-free impedimetric sensing platform for microRNA-21 based on ZrO2-reduced graphene oxide nanohybrids coupled with catalytic hairpin assembly amplification.

Herein, a sensitive electrochemical impedance sensor was constructed based on ZrO2-reduced graphene oxide (RGO)-modified electrode coupled with the catalytic hairpin assembly signal amplification strategy. Electrochemical impedance spectroscopy (EIS) was used to detect microRNA (miRNA) using the change in electron transfer resistance (ΔRet) originated from nucleic acid hybridization on the electrode surface. MiRNA-21 was used as a model to verify this strategy. The results indicated that ΔRet exhibited a good linear relationship with the concentration of miRNA-21 in the range from 1.0 × 10−14 mol L−1 to 1.0 × 10−10 mol L−1 with a detection limit of 4.3 × 10−15 mol L−1 (S/N = 3). Additionally, this sensor exhibited good selectivity, and it could be applied to detect miRNA-21 in human serum samples and measure the expression levels of miRNA-21 in human breast cancer cell lines (MCF-7); thus, this sensor has great potential in cancer diagnosis.

Label-free peptide aptamer based impedimetric biosensor for highly sensitive detection of TNT with a ternary assembly layer

We report a label-free peptide aptamer based biosensor for highly sensitive detection of TNT which was designed with a ternary assembly layer consisting of anti-TNT peptide aptamer (peptamer), dithiothreitol (DTT), and 6-mercaptohexanol (MCH), forming Au/peptamer-DTT/MCH. A linear relationship between the change in electron transfer resistance and the logarithm of the TNT concentration from 0.44 to 18.92 pM, with a detection limit of 0.15 pM, was obtained. In comparison, the detection limit of the aptasensor with a common binary assembly layer (Au/peptamer/MCH) was 0.15 nM. The remarkable improvement in the detection limit could be ascribed to the crucial role of the ternary assembly layer, providing an OH-richer hydrophilic environment and a highly compact surface layer with minimal surface defects, reducing the non-covalent binding (physisorption) of the peptamer and non-specific adsorption of TNT onto the electrode surface, leading to high sensitivity, and which can serve as a general sensing platform for the fabrication of other biosensors.

Anisotropic In Situ-Coated AuNPs on Screen-Printed Carbon Surface for Enhanced Prostate-Specific Antigen Impedimetric Aptasensor

An impedimetric aptasensor has been used to study the effect of charge transfer on the binding of prostate-specific antigen (PSA) to its aptamer. Full understanding of this mechanism will be beneficial to further improve its sensitivity for PSA detection in human semen at physiologically relevant concentrations. Bare gold electrodes (SPAuEs) and gold nanoparticles (AuNPs)-coated screen-printed carbon ink electrodes (AuNPs/SPCEs) were coated with aptamer solution at various concentrations and the sensor response to increasing PSA concentration in buffer solution examined. AuNPs were deposited onto carbon electrodes in 10 cycles. AuNPs/SPCEs were then coated with a self-assembled monolayer (SAM) of 16-mercaptohexadecanoic acid prior to aptamer immobilization at dose of 5 μg mL−1. The results indicate that anisotropic AuNPs/SPCEs outperform bare gold electrodes in terms of decreased amount of aptamer bunches as well as the number of intermediate PSA-aptamer complexes formed on the electrode surface. The key finding is that the fabricated aptasensor is sensitive enough [limit of detection (LoD) 1.95 ng mL−1] for early diagnosis of prostate cancer and displays linear response in the physiologically relevant concentration range (0 ng mL−1 to 10 ng mL−1), as shown by the calibration curve of the relative change in electron transfer resistance (ΔRCT) versus PSA concentration when aptamer/SAM/AuNPs/SPCEs were exposed to buffer containing PSA at different concentrations.

Rapid Quantitative Detection of Brucella melitensis by a Label-Free Impedance Immunosensor Based on a Gold Nanoparticle-Modified Screen-Printed Carbon Electrode

A rapid and simple method for quantitative monitoring of Brucella melitensis using electrochemical impedance spectroscopy (EIS) is reported for the first time. The label-free immunosensors were fabricated by immobilizing Brucella melitensis antibody on the surface of gold nanoparticle-modified screen-printed carbon electrodes (GNP-SPCEs). Cyclic voltammetry (CV) and EIS were used to characterize the Brucella melitensis antigen interaction on the surface of GNP-SPCEs with antibody. A general electronic equivalent model of an electrochemical cell was introduced for interpretation of the impedance components of the system. The results showed that the change in electron-transfer resistance (Rct) was significantly different due to the binding of Brucella melitensis cells. A linear relationship between the Rct variation and logarithmic value of the cell concentration was found from 4 × 104 to 4 × 106 CFU/mL in pure culture. The label-free impedance biosensor was able to detect as low as 1 × 104 and 4 × 105 CFU/mL of Brucella melitensis in pure culture and milk samples, respectively, in less than 1.5 h. Moreover, a good selectivity versus Escherichia coli O157:H7 and Staphylococcus aureus cells was obtained for our developed immunosensor demonstrating its specificity towards only Brucella melitensis.

A rapid electrochemical biosensor based on an AC electrokinetics enhanced immuno-reaction

Fluorescent labelling and chromogenic reactions that are commonly used in conventional immunoassays typically utilize diffusion dominated transport of analytes, which is limited by slow reaction rates and long detection times. By integrating alternating current (AC) electrokinetics and electrochemical impedance spectroscopy (EIS), we construct an immunochip for rapid, sensitive, and label-free detection. AC electroosmosis (ACEO) and positive dielectrophoresis (DEP), induced by a biased AC electric field, can rapidly convect and trap the analyte onto an EIS working electrode within a few minutes. This allows the change of electron-transfer resistance (ΔRet) caused by the antibody-antigen (IgG-protein A) binding to be measured and quantified in real time. The measured impedance change achieves a plateau after electrokinetic concentration for only 90 s, and the detection limit is able to reach 200 pg ml⁻¹. Compared to the conventional incubation method, the electrokinetics-enhanced method is approximately 100 times faster in its reaction time, and the detection limit is reduced by 30 times. The ΔRet of the positive response is two orders of magnitude higher than the negative control, demonstrating excellent specificity for practical applications.

Interdigitated microelectrode-based microchip for electrical impedance spectroscopic study of oral cancer cells

In this study, electric/electrochemical impedance spectroscopy and cyclic voltammetry were used to study the cellular activities of oral cancer cell line CAL 27, including the kinetics of cell adhesion, spreading, and cell proliferation on interdigitated microelectrodes (IMEs). Impedance spectra of CAL 27 cells on IMEs electrodes were obtained in cell growth medium and in 0.1 M PBS with 50 mM [Fe(CN)₆]³⁻/⁴⁻ as redox probe. Equivalent circuits were used to model both cases. In cell growth medium, impedance spectra allowed us to analyze the changes in capacitance and resistance due to cell attachment on the IMEs over the entire experiment period. It was found that cell spreading caused the most significant decrease in capacitance component and slight increase in resistance component. Impedance change at given frequencies (between 10 kHz to 100 kHz) was found to be linearly increased with increasing cell number of CAL 27 on the IMEs. In comparison with non-cancer oral epithelial cells (Het-1A), at equal cell number, cancer cells always generated impedance several folds higher than that of non-cancer cells. In the presence of [Fe(CN)₆]³⁻/⁴⁻, impedance spectra allowed us to analyze the change in electron transfer resistance of IMEs due to cell attachment, in which an increase trend was observed at 24 h with increasing cell number from 2500 cells to 10,000 cells on IMEs. Double layer capacitance was also affected by cell attachment, and a decrease in double layer capacitance was observed with increasing cell number on the electrodes. Cyclic voltammetric measurements correlated well with the impedance results. The results of this study demonstrated the use of electrochemical approaches to obtain and understand cellular behaviors/activities of oral cancer cells, potentially providing useful tools for cancer cell research.

Modelling of the impedimetric responses of an aflatoxin B1 immunosensor prepared on an electrosynthetic polyaniline platform

Aflatoxins are a group of mycotoxins that have deleterious effects on humans and are produced during fungal infection of plants or plant products. An electrochemical immunosensor for the determination of aflatoxin B(1) (AFB(1)) was developed with AFB(1)antibody (AFB(1)-Ab) immobilized on Pt electrodes modified with polyaniline (PANi) and polystyrene sulphonic acid (PSSA). Impedimetric analysis shows that the electron transfer resistances of the Pt/PANi-PSSA electrode, the Pt/PANi-PSSA/AFB(1)-Ab immunosensor and Pt/PANi-PSSA/AFB(1)-Ab incubated in bovine serum albumin (BSA) were 0.458, 720 and 1,066 kOmega, respectively. These results indicate that electrochemical impedance spectroscopy (EIS) is a suitable method for monitoring the change in electron transfer resistance associated with the immobilization of the antibody. Modelling of EIS data gave equivalent circuits which showed that the electron transfer resistance increased from 0.458 kOmega for the Pt/PANi-PSSA electrode to 1,066 kOmega for the Pt/PANi-PSSA/AFB(1)-Ab immunosensor, indicating that immobilization of the antibody and incubation in BSA introduced an electron transfer barrier. The AFB(1) immunosensor had a detection limit of 0.1 mg/L and a sensitivity of 869.6 kOmega L/mg.

Electrochemical and piezoelectric quartz crystal detection of antisperm antibody based on protected Au nanoparticles with a mixed monolayer for eliminating nonspecific binding

The detection of antisperm antibody (AsAb) by electrochemical method based on Au nanoparticles with a mixed monolayer for eliminating nonspecific binding is presented. Impedance spectroscopy is used to characterize the modified procedures, the immobilization of sperm antigen (SpAg) and binding of AsAb in the presence of [Fe (CN) 6] 3−/4−. In addition, piezoelectric quartz crystal, PQC, was used to analyze the immobilization quantity of Au nanoparticles as well as to optimize experimental conditions. The change of electron-transfer resistance was correlated with the concentration of AsAb in a range from 25 to 600 mU/ml with the detection limit of 10 mU/ml. The analytical results of several human serum samples using the developed technique are in satisfactory agreement with those given by the enzyme-linked immunosorbent assay (ELISA) method.

The Novel Immunobiosensors for Detection of Escherichia coli O157:H7 Using Electrochemical Impedance Spectroscopy.

Immunobiosensors were developed for detection of Escherichia coli O157:H7 based on the surface immobilization of monoclone antibodies onto indium tin oxide (ITO) electrodes. The immobilization of antibodies onto ITO chips was carried out by silanization. The effects of epoxysilane monolayer, the antibody layer on the electrochemical properties of the electrode, and the combined target bacteria were analyzed through cyclic voltammetry and electrochemical impedance spectroscopy. By using Randles model as the equivalent circuit, the concentration of the target bacteria can be quantitatively analyzed in terms of the change of electron transfer resistance. The biosensor could detect the target bacteria with a detection limit of 4×103CFU/mL. A linear response was found between 4×103- 4×106CFU/mL. This biosensor was characterized with high sensitivity, excellent selectivity, short detection time and easy operation It has a promising application in clinical laboratory diagnoses, environmental detection and food safety.