Glucose biosensing based on the highly efficient immobilization of glucose oxidase on a Prussian blue modified nanostructured Au surface

We report on a novel glucose biosensor based on the immobilization of glucose oxidase (GOx) on a Prussian blue modified nanoporous gold surface. The amperometric glucose biosensor fabricated in this study exhibits a fast response and the very low detection limit of 2.5 μM glucose. The sensitivity of the biosensor was found to be very high, 177 μA/mM; the apparent Michaelis–Menten constant is calculated to be 2.1 mM. In addition, the biosensor has good reproducibility and remains stable over 60 days. The anti-interference ability of the biosensor was also assessed, showing little interference from possible interferents such as ascorbic acid (AA), acetaminophen (AP) and uric acid (UA).     

Comparative studies on electrogenerated chemiluminescence of luminol on gold nanoparticle modified electrodes

Comparative studies on the electrogenerated chemiluminescence (ECL) behavior of luminol on various electrodes modified with gold nanoparticles of different size were carried out in neutral solution by conventional cyclic voltammetry (CV). The results demonstrated that the gold nanoparticle modified electrodes could generate strong luminol ECL in neutral pH conditions. The catalytic performance of gold nanoparticle modified electrodes on luminol ECL depended not only on the gold nanoparticles but also on the substrate. Gold electrode and glassy carbon electrode were the most suitable substrates for the self-assembly of gold nanoparticles. Moreover, the gold nanoparticle modified gold and glassy carbon electrode had satisfying stability and reproducibility and did not need tedious pretreatment of electrode surface before each measurement. It was also found that luminol ECL behavior depended on the size of gold nanoparticles. The most intense ECL signals were obtained on a 16-nm-diameter gold nanoparticle modified electrode. The modified electrode prepared by the self-assembly method exhibited much better catalytic effect on luminol ECL than that prepared by the electrically deposited method. The ECL behavior of luminol on a gold nanoparticle self-assembled gold electrode was also investigated by other transient-state electrochemical techniques, such as chronoamperometry, differential pulse voltammetry, normal pulse voltammetry, and square wave voltammetry. The strongest ECL intensity was obtained under square wave voltammetric condition.     

Enzyme immobilisation on electroactive nanostructured membranes (ENM): optimised architectures for biosensing

Electroactive nanostructured membranes have been produced by the layer-by-layer (LbL) technique, and used to make electrochemical enzyme biosensors for glucose by modification with cobalt hexacyanoferrate redox mediator and immobilisation of glucose oxidase enzyme. Indium tin oxide (ITO) glass electrodes were modified with up to three bilayers of polyamidoamine (PAMAM) dendrimers containing gold nanoparticles and poly(vinylsulfonate) (PVS). The gold nanoparticles were covered with cobalt hexacyanoferrate that functioned as a redox mediator, allowing the modified electrode to be used to detect H₂O₂, the product of the oxidase enzymatic reaction, at 0.0 V vs. SCE. Enzyme was then immobilised by cross-linking with glutaraldehyde. Several parameters for optimisation of the glucose biosensor were investigated, including the number of deposited bilayers, the enzyme immobilisation protocol and the concentrations of immobilised enzyme and of the protein that was crosslinked with PAMAM. The latter was used to provide glucose oxidase with a friendly environment, in order to preserve its bioactivity. The optimised biosensor, with three bilayers, has high sensitivity and operational stability, with a detection limit of 6.1 μM and an apparent Michaelis–Menten constant of 0.20 mM. It showed good selectivity against interferents and is suitable for glucose measurements in natural samples.     

Electrochemiluminescence biosensor for carcinoembryonic antigen detection based on Au-Ag/g-C3N4 nanocomposites

Herein, an electrochemiluminescence (ECL) aptasensor for carcinoembryonic antigen (CEA) detection was developed based on Au-Ag/g-C₃N₄ nanocomposites (NCs), which were synthesized by decorating graphitic carbon nitride (g-C₃N₄) nanosheets with alloy-structured Au-Ag bimetallic nanoparticles (NPs) via one-step in situ chemical reduction. As ECL sensing platform, Au-Ag/g-C₃N₄ NCs could significantly improve the ECL intensity of luminol due to the good conductivity of Au-Ag NPs, electrocatalytic activity for oxygen evolution reaction (OER) and the ability to adsorb luminol via π stacking interaction. In addition, it could load the thiol terminated aptamers of CEA via Au-S or Ag-S bonds. In the presence of CEA, the ECL response of the proposed biosensor decreased significantly due to the fact that the assembled protein layers hindered the electron transfer and the diffusion of ECL reactants toward the electrode surface. The proposed ECL sensor exhibited a good linear relationship with CEA in the range of 1.0–1.0 × 10^?6 ng/mL with a detection limit of 8.9 × 10^?7 ng/mL. The satisfactory results were obtained in the detection of CEA in human serum samples.     

Amperometric glucose biosensor based on a gold nanorods/cellulose acetate composite film as immobilization matrix

We report on the utilization of gold nanorods to create a highly responsive glucose biosensor. The feasibility of an amperometric glucose biosensor based on immobilization of glucose oxidase (GOx) in gold nanorod is investigated. GOx is simply mixed with gold nanorods and cross-linked with a cellulose acetate (CA) medium by glutaraldehyde. The adsorption of GOx on the gold nanorods is confirmed by X-ray photoelectron spectroscopy (XPS) measurements. Circular dichroism (CD) and UV-spectrum results show that the activity of GOx was preserved after conjugating with gold nanorods. The current response of modified electrode is 10 times higher than that of without gold nanorods. Under optimal conditions, the biosensor shows high sensitivity (8.4 μA cm⁻² mM⁻¹), low detection limit (2 × 10⁻⁵ M), good storage stability and high affinity to glucose (). A linear calibration plot is obtained in the wide concentration range from 3 × 10⁻⁵ to 2.2 × 10⁻³ M.     

An ultrasensitive luminol cathodic electrochemiluminescence immunosensor based on glucose oxidase and nanocomposites: Graphene–carbon nanotubes and gold-platinum alloy

In the present study, a novel and ultrasensitive electrochemiluminescence (ECL) immunosensor based on luminol cathodic ECL was fabricated by using Au nanoparticles and Pt nanoparticles (nano-AuPt) electrodeposited on graphene–carbon nanotubes nanocomposite as platform for the detection of carcinoembryonic antigen (CEA). For this introduced immunosensor, graphene (GR) and single wall carbon nanotubes (CNTs) dispersed in chitosan (Chi-GR-CNTs) were firstly decorated on the bare gold electrode (GE) surface. Then nano-AuPt were electrodeposited (DpAu-Pt) on the Chi-GR-CNTs modified electrode. Subsequently, glucose oxidase (GOD) was employed to block the non-specific sites of electrode surface. When glucose was present in the working buffer solution, GOD immediately catalyzed the oxidation of glucose to in situ generate hydrogen peroxide (H₂O₂), which could subsequently promote the oxidation of luminol with an amplified cathodic ECL signal. The proposed immunosensor was performed at low potential (−0.1 to 0.4 V) and low concentration of luminol. The CEA was determined in the range of 0.1 pg mL⁻¹ to 40 ng mL⁻¹ with a limit of detection down to 0.03 pg mL⁻¹ (S N⁻¹ = 3). Moreover, with excellent sensitivity, selectivity, stability and simplicity, the as-proposed luminol-based ECL immunosensor provided great potential in clinical applications.     

A novel electrochemiluminescence glucose biosensor based on platinum nanoflowers/graphene oxide/glucose oxidase modified glassy carbon electrode

A novel electrochemiluminescence (ECL) biosensor based on platinum nanoflowers (PtNFs)/graphene oxide (GO)/glucose oxidase (GODx) was discovered for glucose detection. PtNFs/GO was synthesized using a nontoxic, rapid, one-pot and template-free method and characterized by transmission electron microscopy (TEM) and high-resolution TEM techniques. The as-prepared PtNFs/GO with clean surface and multiporous structure was used to assemble GODx to form a glucose biosensor. Based on ECL results, the PtNFs/GO/GODx film-modified electrode displayed a high electrocatalytic activity towards the oxidation of glucose, which generated hydrogen peroxide (H₂O₂) to react with the luminol radicals thus enhanced the luminol ECL. Under the optimized conditions, two linear regions of ECL intensity to glucose concentration were valid in the range from 5 to 80 μmol/L (r?=?0.9957) and 80 to 1,000 μmol/L (r?=?0.9909) with a detection limit (S/N?=?3) of 2.8 μmol/L. In order to verify the reliability, the thus-fabricated biosensor was applied to determine the glucose concentration in glucose injection, glucose functional drink, and blood serum. The results indicated that the proposed biosensor presented good characteristics in terms of high sensitivity and good reproducibility for glucose determination, promising the applicability of this sensor in practical analysis.     

基于碳纳米管修饰电极的胆碱电化学发光生物传感器研制

在碳纳米管(CNTs)和K3Fe(CN)6修饰的铂电极上吸附固定胆碱氧化酶,以鲁米诺为发光试剂,研制了胆碱电化学发光(ECL)生物传感器.CNTs可有效提高电极表面的电荷传输能力、提高电极表面的生物相容性和对酶分子的固载能力;K3Fe(CN)6对酶活性具有激活作用,同时对H2O2增敏的鲁米诺ECL有增强作用,均有利于提...     

Determination of cocaine on banknotes through an aptamer-based electrochemiluminescence biosensor

A novel electrochemiluminescence (ECL) “sandwich” biosensor has been developed to detect cocaine. The sandwich biosensor was fabricated on the basis of the fact that a single aptamer could be split into two fragments and the two dissociated parts could form a folded, associated complex in the presence of targets. One of these (capture probe), which had hexane–thiol at its 5′-terminus, was immobilized on a gold electrode via thiol–gold binding. The other one (detection probe) was labeled with the ECL reagent tris(2,2′-bipyridyl)ruthenium(II)-doped silica nanoparticles (RuSiNPs) at its 3′-terminus. Owing to the weak interaction between the two fragments, the sensor exhibited a low ECL signal in the absence of cocaine. After the target cocaine had been added to the solution, it induced association of the two fragments and stabilized the associated complexes, leading to immobilization of RuSiNPs on the electrode surface, and the ECL detected on the electrode surface was enhanced. The enhanced ECL intensity was directly proportional to the logarithm of the cocaine concentration in the range from 1.0 × 10⁻⁹ to1.0 × 10⁻¹¹ mol/L, with a detection limit of 3.7 × 10⁻¹² mol/L. The biosensor was applied to detect trace amounts of cocaine on banknotes with satisfactory results.     

Electrogenerated chemiluminescence of luminol on a gold-nanorod-modified gold electrode

Electrogenerated chemiluminescence (ECL) of luminol on a gold-nanorod-modified gold electrode was studied, and five ECL peaks were obtained under conventional cyclic voltammetry in both neutral and alkaline solutions. Among them, four ECL peaks (ECL-1-4) were also observed on a gold-nanosphere-modified gold electrode, but the intensities of these ECL peaks were enhanced about 2-10-fold on a gold-nanorod-modified gold electrode in neutral solution. One new strong ECL peak (ECL-5) was obtained at -0.28 V (vs SCE) on a gold-nanorod-modified gold electrode in both neutral and alkaline solutions and enhanced with an increase in pH. In strong alkaline solutions, ECL-1 and ECL-2 on a gold-nanosphere-modified electrode were much stronger than those on a gold-nanorod-modified gold electrode, while ECL-3-5 appeared to only happen on a gold-nanorod-modified gold electrode. The emitter of all the ECL peaks was identified as 3-aminophthalate. The ECL peaks were found to depend on the scan direction, the electrolytes, the pH, and the presence of O(2) and N(2). The reaction pathways for ECL-4 have been further elucidated, and the mechanism of the new ECL peak (ECL-5) has been proposed. The results indicate that a gold-nanorod-modified gold electrode has a catalytic effect on luminol ECL different from that of a gold-nanosphere-modified gold electrode, revealing that the shape of the metal nanoparticles has an important effect on the luminol ECL behavior. The strong ECL of luminol in neutral solution obtained on a gold-nanorod-modified electrode may be used for the sensitive detection of biologically important compounds in physiological conditions.     

Effect of gold nanoparticle as a novel nanocatalyst on luminol-hydrazine chemiluminescence system and its analytical application

In this work the catalytic role of unsupported gold nanoparticles on the luminol–hydrazine reaction is investigated. Gold nanoparticles catalyze the reaction of hydrazine and dissolved oxygen to generate hydrogen peroxide and also catalyze the oxidation of luminol by the produced hydrogen peroxide. The result is an intense chemiluminescence (CL) due to the excited 3-aminophthalate anion. In the absence of gold nanoparticles no detectable CL was observed by the reaction of luminol and hydrazine unless an external oxidant is present in the system. The size effect of gold nanoparticles on the CL intensity was investigated. The most intensive CL signals were obtained with 15-nm gold nanoparticles. UV–vis spectra and transmission electron microscopy studies were used to investigate the CL mechanism. The luminol and hydroxide ion concentration, gold nanoparticles size and flow rate were optimized. The proposed method was successfully applied to the determination of hydrazine in boiler feed water samples. Between 0.1 and 30 μM of hydrazine could be determined with a detection limit of 30 nM.     

Direct electrochemistry of glucose oxidase based on its direct immobilization on carbon ionic liquid electrode and glucose sensing

Direct electrochemistry of glucose oxidase (GOx) has been achieved by its direct immobilization on carbon ionic liquid electrode (CILE) with a conductive hydrophobic ionic liquid, 1-butyl pyridinium hexafluophosphate ([BuPy][PF₆]) as binder for the first time. A pair of reversible peaks is exhibited on GOx/CILE by cyclic voltammetry. The peak-to-peak potential separation (ΔE_P) of immobilized GOx is 0.056 V in 0.067 M phosphate buffer solution (pH 6.98) with scan rate of 0.1 V/s. The average surface coverage and the apparent Michaelis–Menten constant are 6.69 × 10⁻¹¹ mol·cm⁻² and 2.47 μM. GOx/CILE shows excellent electrocatalytic activity towards glucose determination in the range of 0.1–800 μM with detection limit of 0.03 μM (S/N = 3). The biosensor has been successfully applied to the determination of glucose in human plasma with the average recoveries between 95.0% and 102.5% for three times determination. The direct electrochemistry of GOx on CILE is achieved without the help of any supporting film or any electron mediator. GOx/CILE is inexpensive, stable, repeatable and easy to be fabricated.     

Electrogenerated chemiluminescence aptamer-based biosensor for the determination of cocaine

A novel electrogenerated chemiluminescence aptamer-based (ECL-AB) biosensor for the determination of a small molecule drug is designed employing cocaine-binding aptamer as molecular recognition element for cocaine as a model analyte and ruthenium complex served as an ECL label. A 5′-terminal cocaine-binding aptamer with the ECL label at 3′-terminal of the aptamer was utilized as an ECL probe. The ECL-AB biosensors were fabricated by immobilizing the ECL probe onto a gold electrode surface via thiol-Au interactions. An enhanced ECL signal is generated upon recognition of the target cocaine, attributed to a change in the conformation of the ECL probe from random coil-like configuration on the probe-modified film to three-way junction structure, in close proximity to the sensor interface. The integrated ECL intensity versus the concentration of cocaine was linear in the range from 5.0 × 10⁻⁹ to 3.0 × 10⁻⁷ M. The detection limit was 1.0 × 10⁻⁹ M. This work demonstrates that the combination of a highly binding aptamer to analyte with a highly sensitive ECL technique to design ECL-AB biosensor is a great promising approach for the determination of small molecule drugs.     

Electrochemiluminescence biosensor for the assay of small molecule and protein based on bifunctional aptamer and chemiluminescent functionalized gold nanoparticles

An electrochemiluminescence (ECL) biosensor for simultaneous detection of adenosine and thrombin in one sample based on bifunctional aptamer and N-(aminobutyl)-N-(ethylisoluminol) functionalized gold nanoparticles (ABEI-AuNPs) was developed. A streptavidin coated gold nanoparticles modified electrode was utilized to immobilize biotinylated bifunctional aptamer (ATA), which consisted of adenosine and thrombin aptamer. The ATA performed as recognition element of capture probe. For adenosine detection, ABEI-AuNPs labeled hybridization probe with a partial complementary sequence of ATA reacted with ATA, leading to a strong ECL response of N-(aminobutyl)-N-(ethylisoluminol) enriched on ABEI-AuNPs. After recognition of adenosine, the hybridization probe was displaced by adenosine and ECL signal declined. The decrease of ECL signal was in proportion to the concentration of adenosine over the range of 5.0 × 10⁻¹²–5.0 × 10⁻⁹ M with a detection limit of 2.2 × 10⁻¹² M. For thrombin detection, thrombin was assembled on ATA modified electrode via aptamer–target recognition, another aptamer of thrombin tagged with ABEI-AuNPs was bounded to another reactive site of thrombin, producing ECL signals. The ECL intensity was linearly with the concentration of thrombin from 5 × 10⁻¹⁴ M to 5 × 10⁻¹⁰ M with a detection limit of 1.2 × 10⁻¹⁴ M. In the ECL biosensor, adenosine and thrombin can be detected when they coexisted in one sample and a multi-analytes assay was established. The sensitivity of the present biosensor is superior to most available aptasensors for adenosine and thrombin. The biosensor also showed good selectivity towards the targets. Being challenged in real plasma sample, the biosensor was confirmed to be a good prospect for multi-analytes assay of small molecules and proteins in biological samples.     

Highly sensitive electrogenerated chemiluminescence biosensor in profiling protein kinase activity and inhibition using a multifunctional nanoprobe

We presented a novel electrogenerated chemiluminescence (ECL) biosensor for monitoring the activity and inhibition of protein kinases based on signal amplification using enzyme-functionalized Au NPs nanoprobe. In this design, the biotin-DNA labeled glucose oxidase/Au NPs (GOx/Au NPs/DNA-biotin) nanoprobes, prepared by conjugating Au NPs with biotin-DNA and GOx, were bound to the biotinylated anti-phosphoserine labeled phosphorylated peptide modified electrode surface through a biotin−avidin interaction. The GOx assembled on the nanoprobe can catalyze glucose to generate H₂O₂ in the presence of O₂ while the ECL reaction occurred in the luminol ECL biosensor. At a higher concentration of kinase, there are more nanoprobes on the electrode, which gives a higher amount of GOx at the electrode interface and thus higher electrocatalytic efficiency to the luminol ECL reaction. Therefore, the activity of protein kinases can be monitored by ECL with high sensitivity. Protein kinase A (PKA), an important enzyme in regulation of glycogen, sugar, and lipid metabolism in the human body, was used as a model to confirm the present proof-of-concept strategy. The as-proposed biosensor presents high sensitivity, low detection limit of 0.013 U mL⁻¹, wide linear range (from 0.02 to 40 U mL⁻¹), and excellent stability. Moreover, this biosensor can also be used for quantitative analysis of kinase inhibition. On the basis of the inhibitor concentration dependent ECL signal, the half-maximal inhibition value IC₅₀ of ellagic acid, a typical PKA inhibitor, was estimated, which is in agreement with those obtained using the conventional kinase assay. The simple and sensitive biosensor is promising in developing a high-through assay of in vitro kinase activity and inhibitor screening for clinic diagnostic and drug development.     

基于工具酶信号放大技术电化学发光法检测凝血酶

利用巯基乙胺将合成的金纳米粒子氨基化;基于纳米粒子负载羧基化的联吡啶钌和巯基DNA制得电化学发光信号探针;采用酶循环信号放大技术,获得大量含新增DNA的溶液来捕获信号探针;以金电极为载体,将巯基DNA自组装到电极表面,依次杂交互补DNA和信号探针,构建电化学发光生物传感器.在优化的条件下,此传感器对凝血酶具有良好的响应,在3.0× 10-13～6.0×10-11 mol/L范围内,凝血酶的浓度与发光强度呈良好的线性关系,检出限为1.8× 10-13 mol/L(3a).采用酶切循环放大技术制备的生物传感器具有灵敏度高,选择性和重现性良好等特点.     

A Highly Sensitive Electrochemiluminescence Choline Biosensor Based on Poly(aniline‐luminol‐hemin) Nanocomposites

Poly(aniline‐luminol‐hemin) nanocomposites are prepared on an electrode surface through electropolymerization, and a highly sensitive electrochemiluminescence (ECL) biosensor for choline is developed based on the poly(aniline‐luminol‐hemin) nanocomposites and an enzyme catalyzed reaction of choline oxidase (CHOD). The obtained nanocomposites are characterized by scanning electron microscopy (SEM), atomic absorption spectrometry (AAS) and ECL. The results indicate that hemin can be incorporated into the poly(aniline‐luminol) nanocomposites using the facile electropolymerization method, and the poly(aniline‐luminol‐hemin) nanocomposites are rod shaped porous nanostructure. Moreover, the poly(aniline‐luminol‐hemin) nanocomposites exhibit higher ECL intensity than poly(aniline‐luminol) nanocomposites in alkaline media due to the catalytic effect of hemin on the ECL of the polymerized luminol and the electron transfer ability of hemin in the nanocomposites. CHOD is immobilized on the surface of the poly(aniline‐luminol‐hemin) nanocomposites modified electrode with glutaraldehyde, and the ECL biosensor based on poly(aniline‐luminol‐hemin)/CHOD exhibits a wider linear range for the choline detection. The enhanced ECL signals are linear with the logarithm of concentration of choline over the range of 1.0×10^?11～1.0×10^?7 mol L^?1 with a low detection limit of 1.2×10^?12 mol L^?1. Moreover, the proposed biosensor is successfully applied to the detection of choline in milk.     

Disposable luminol copolymer-based biosensor for uric acid in urine

A new electrochemiluminescent (ECL) disposable biosensor for uric acid was manufactured by immobilization in a double-layer design of luminol as a copolymer with 3,3′,5,5′-tetramethylbenzidine (TMB) and the enzyme uricase in chitosan on gold screen-printed cells. The good mechanical and improved electroluminescent characteristics of the new copolymer poly(luminol–TMB) make it possible to determine uric acid by measuring the growing ECL emission with the analyte concentration. The combination of enzymatic selectivity with ECL sensitivity results in a disposable analytical device with a linear range for uric acid from 1.5 × 10⁻⁶ to 1.0 × 10⁻⁴ M, a limit of detection of 4.4 × 10⁻⁷ M and a precision of 13.1% (1.0 × 10⁻⁵ M, n = 10) as relative standard deviation. Satisfactory results were obtained for uric acid determination in 24 h-urine samples compared to a reference procedure. This uric acid biosensor can be used as a low-cost alternative to conventional methods.     

Enhanced electrochemiluminescence from luminol at multi-walled carbon nanotubes decorated with palladium nanoparticles: a novel route for the fabrication of an oxygen sensor and a glucose biosensor

Incorporation of palladium nanoparticles on the surface of multi-walled carbon nanotubes and modification of glassy carbon electrode with the prepared nano-hybrid material led to the fabrication of a novel electrode. The modified electrode showed attractive electrocatalytic activity and sensitizing effect on luminol-O(2) and luminol-H(2)O(2) electrochemiluminescence (ECL) reactions at neutral media. The sensitized luminol-O(2) and luminol-H(2)O(2) reactions were successfully applied for the ECL determination of dissolved O(2) and glucose, respectively. Under the optimal conditions for luminol-O(2) system, the ECL signal intensity of luminol was linear with the concentration of dissolved oxygen in the range between 0.08 and 0.94 mM (r=0.9996) and for luminol-H(2)O(2) system, the ECL signal intensity of luminol was linear with the concentration of glucose in the range between 0.1 and 1000 μM (r=0.9998). The limits of detection (S/N=3) for dissolved oxygen and glucose were 0.02 mM and 54 nM, respectively. The relative standard deviations (RSD) for repetitive measurements of 0.50 mM oxygen (n=10) and 10 μM glucose (n=30) were 3.5% and 0.3%, respectively. Also, under the optimal conditions for luminol-H(2)O(2) system, the ECL signal intensity of luminol was linear with the concentration of H(2)O(2) in the range between 1 nM and 0.45 mM (r=0.9997). The limit of detection (S/N=3) for H(2)O(2) detection was 0.5 nM and the relative standard deviation for repetitive measurements of 10 μM H(2)O(2) (n=10) was 0.8%.     

Synthesis, characterization, and electrochemiluminescence of luminol-reduced gold nanoparticles and their application in a hydrogen peroxide sensor

It was found that chloroauric acid (HAuCl(4)) could be directly reduced by the luminescent reagent luminol in aqueous solution to form gold nanoparticles (AuNPs), the size of which depended on the amount of luminol. The morphology and surface state of as-prepared AuNPs were characterized by transmission electron microscopy, UV/visible spectroscopy, X-ray photoelectron spectroscopy, FTIR spectroscopy, and thermogravimetric analysis. All results indicated that residual luminol and its oxidation product 3-aminophthalate coexisted on the surface of AuNPs through the weak covalent interaction between gold and nitrogen atoms in their amino groups. Subsequently, a luminol-capped AuNP-modified electrode was fabricated by the immobilization of AuNPs on a gold electrode by virtue of cysteine molecules and then immersion in a luminol solution. The modified electrode was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, and scanning electron microscopy. The as-prepared modified electrode exhibited an electrochemiluminescence (ECL) response in alkaline aqueous solution under a double-step potential. H2O2 was found to enhance the ECL. On this basis, an ECL sensor for the detection of H2O2 was developed. The method is simple, fast, and reagent free. It is applicable to the determination of H2O2 in the range of 3x10(-7)-1x10(-3) mol L(-1) with a detection limit of 1x10(-7) mol L(-1) (S/N=3).