Preparation of multiwalled carbon nanotubes-platinum-Nafion-glucose oxidase nanobiocomposite and its application to glucose biosensor

Platinum (Pt) nanoparticles were electrodeposited within multiwalled carbon nanotubes-Nafion-glucose oxidase (MWNTs-Nafion-GOx) nanobiocomposite by a potentiostatic method. The morphology and nature of the resulting MWNTs-Pt-Nafion-GOx nanobiocomposite were characterized by field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), and energy dispersive X-ray spectroscopy (EDS). The electrocatalytic properties of the MWNTs-Pt-Nafion-GOx nanobiocomposite film modified glassy carbon electrode were characterized by cyclic voltammetry and amperometry in the presence of hydrogen peroxide. The glucose biosensor sensitivity was strongly influenced by the deposits of Pt nanoparticles and amount of GOx concentration within the MWNTs-Pt-Nafion-GOx nanobiocomposite film. The optimized glucose biosensor displayed a sensitivity of 640 nA mM(-1), a linear range of up to 4 mM, a detection limit of 4 microM, and a response time of less than 4 s at an operating potential of +500 mV versus Ag/AgCl (3 M KCl).     

DNA as a support for glucose oxidase immobilization at Prussian blue-modified glassy carbon electrode in biosensor preparation

An amperometric glucose biosensor has been developed using DNA as a matrix of Glucose oxidase (GOx) at Prussian-blue (PB)-modified glassy carbon (GC) electrode. GC electrode was chemically modified by the PB. GOx was immobilized together with DNA at the working area of the PB-modified electrode by placing a drop of the mixture of DNA and GOx. The response of the biosensor for glucose was evaluated amperometrically. Upon immobilization of glucose oxidase with DNA, the biosensor showed rapid response toward the glucose. On the other hand, no significant response was obtained in the absence of DNA. Experimental conditions influencing the biosensor performance were optimized and assessed. This biosensor offered an excellent electrochemical response for glucose concentration in micro mol level with high sensitivity and selectivity and short response time. The levels of the relative standard deviation (RSDs), (<4%) for the entire analyses reflected a highly reproducible sensor performance. Through the use of optimized conditions, a linear relationship between current and glucose concentration was obtained up to 4 x 10(-4) M. In addition, this biosensor showed high reproducibility and stability.     

用单壁纳米碳管制备葡萄糖生物传感器的研究

以单壁纳米碳管为代表材料,对利用纳米碳管制备葡萄糖生物传感器中纳米碳管的作用和纳米碳管修饰电极的方法、酶的固定化方法及电极种类等因素对传感器性能的影响进行了研究.研究结果表明,纳米碳管的加入能有效地改善传感器的电化学性能,利用二茂铁和单壁纳米碳管共同修饰电极所制得的传感器的性能要好于仅用单壁纳米碳管修饰电极制得的传感器.在酶的固定化方法中,戊二醛交联法要略好于明胶包埋法;而利用铂电极制备出的生物传感器对葡萄糖的响应电流要明显高于利用金电极和玻碳电极制备出的生物传感器.这些结论对于开发纳米碳管在生物传感领域及生命科学相关领域的应用有参考价值.     

Amperometric biosensor based on carbon nanotubes coated with polyaniline/dendrimer-encapsulated Pt nanoparticles for glucose detection

A novel amperometric glucose biosensor based on the nanocomposites of multi-wall carbon nanotubes (CNT) coated with polyaniline (PANI) and dendrimer-encapsulated Pt nanoparticles (Pt-DENs) is prepared. CNT coated with protonated PANI is in situ synthesized and Pt-DENs is absorbed on PANI/CNT composite surface by self-assembly method. Then Glucose oxidase (GOx) is crosslink-immobilizated onto Pt-DENs/PANI/CNT composite film. The results show that the fabricated GOx/Pt-DENs/PANI/CNT electrode exhibits excellent response performance to glucose, such as low detection limit (0.5 µM), wide linear range (1 µM–12 mM), short response time (about 5 s), high sensitivity (42.0 µA mM^? 1 cm^? 2) and stability (83% remains after 3 weeks).     

Development of bimetal-grown multi-scale carbon micro-nanofibers as an immobilizing matrix for enzymes in biosensor applications

This study describes the development of a novel bimetal (Fe and Cu)-grown hierarchical web of carbon micro-nanofiber-based electrode for biosensor applications, in particular to detect glucose in liquids. Carbon nanofibers (CNFs) are grown on activated carbon microfibers (ACFs) by chemical vapor deposition (CVD) using Cu and Fe as the metal catalysts. The transition metal-fiber composite is used as the working electrode of a biosensor applied to detect glucose in liquids. In such a bi-nanometal-grown multi-scale web of ACF/CNF, Cu nanoparticles adhere to the ACF-surface, whereas Fe nanoparticles used to catalyze the growth of nanofibers attach to the CNF tips. By ultrasonication, Fe nanoparticles are dislodged from the tips of the CNFs. Glucose oxidase (GOx) is subsequently immobilized on the tips by adsorption. The dispersion of Cu nanoparticles at the substrate surface results in increased conductivity, facilitating electron transfer from the glucose solution to the ACF surface during the enzymatic reaction with glucose. The prepared Cu-ACF/CNF/GOx electrode is characterized for various surface and physicochemical properties by different analytical techniques, including scanning electron microscopy (SEM), electron dispersive X-ray analysis (EDX), Fourier-transform infrared spectroscopy (FTIR), BET surface area analysis, and transmission electron microscopy (TEM). The electrochemical tests show that the prepared electrode has fast response current, electrochemical stability, and high electron transfer rate, corroborated by CV and calibration curves. The prepared transition metal-based carbon electrode in this study is cost-effective, simple to develop, and has a stable immobilization matrix for enzymes.     

Decoration of single-walled carbon nanotubes with Pt nanoparticles from an organometallic precursor

Carbon nanotubes (CNTs) are nanomaterials of high interest due to their unique structural, electrical, and mechanical properties. Carbon materials have been widely employed to support metallic nanoparticles for catalysis and electrochemical applications. In this work, we investigated the synthesis of platinum nanoparticles generated from the complex Pt₂(dba)₃ (tris(dibenzylideneacetone) diplatinum) and stabilized with a long alkyl chain amine, hexadecylamine (HDA) and supported on functionalized single-walled carbon nanotubes (SWCNTs). High resolution transmission electron microscopy (HRTEM) studies revealed isolated Pt nanoparticles (2?C3 nm) on SWCNTs. Powder X-ray diffraction (XRD) was used to assess the structure of Pt nanoparticles dispersed on SWCNTs assigned to Pt face-centered cubic (fcc). Additionally, infrared Fourier transform spectroscopy confirmed the presence of the stabilizer at the surface of the Pt nanoparticles even after the purification step and functional groups at the surface of pre-treated SWCNTs. This synthetic method may be an alternative route to prepare small size Pt nanoparticles supported on functionalized SWCNTs.     

Following the biocatalytic activities of glucose oxidase by electrochemically cross-linked enzyme-Pt nanoparticles composite electrodes

An integrated platinum nanoparticles (NPs)/glucose oxidase (GOx) composite film associated with a Au electrode is used to follow the biocatalytic activities of the enzyme. The film is assembled on a Au electrode by the electropolymerization of thioaniline-functionalized Pt NPs and thioaniline-modified GOx. The resulting enzyme/Pt NPs-functionalized electrode stimulates the O 2 oxidation of glucose to gluconic acid and H 2O 2. The modified electrode is then implemented to follow the activity of the enzyme by the electrochemical monitoring of the generated H 2O 2. The effect of the composition of the Pt NPs/GOx cross-linked nanostructures and the optimal conditions for the preparation of the electrodes are discussed.     

Layer-by-layer assemblies of chitosan/multi-wall carbon nanotubes and glucose oxidase for amperometric glucose biosensor applications

A novel amperometric glucose biosensor based on multilayer films containing chitosan, multi-wall carbon nanotubes (MWCNTs) and glucose oxidase (GOD) was developed. MWCNTs were solubilized in chitosan (Chit-MWCNTs) used to interact with GOD. Poly (allylamine) (PAA) and polyvinylsulfuric acid potassium salt (PVS) were alternately deposited on the cleaned Pt electrode surface ((PVS/PAA)₃/Pt). The (PVS/PAA)₃/Pt electrode was alternately immersed in Chit-MWCNTs and GOD to assemble different layers of multilayer films. PBS washing was applied at the end of each assembly deposition for dissociating the weak adsorption. Micrographs of MWCNTs were obtained by scanning electron microscope, and properties of the resulting biosensors were measured by electrochemical measurements. Among the resulting biosensors, the biosensor based on eight layers of multilayer films was best. The resulting biosensor was able to efficiently monitor glucose, with the response time within 8 s, a detection limit of 21 μM estimated at a signal-to-noise ratio of 3, a linear range of 1–10 mM, the sensitivity of 0.45 μA/mM, and well stability. The study can provide a feasible simple approach on developing a new immobilization matrix for biosensors and surface functionalization.     

Multilayer membranes via layer-by-layer deposition of glucose oxidase and Au nanoparticles on a Pt electrode for glucose sensing

We prepared multilayer membranes by the layer-by-layer deposition of glucose oxidase (GOx) and Au nanoparticles (5, 10, or 50 nm φ) on sensor substrates, such as a Pt electrode and a quartz glass plate, to prepare glucose sensors. The enzyme activity of GOx remained even in alternate assemblies, and the activity increased with the increasing number of depositions. The apparent K_m values of the deposited GOx were 28–32 mM, while a reported value in a solution is 33 mM. These results suggest that Au nanoparticles can be used as binders for the deposition of GOx without significant change in the affinity between GOx and glucose.     

Gold nanoparticle-polyaniline composite films for glucose sensing

Gold nanoparticles (AuNPs) have been self-assembled onto electrochemically deposited polyaniline (PANI) films on indium-tin-oxide (ITO) coated glass plates to fabricate glucose biosensor. The covalent immobilization of glucose oxidase (GOx) in the near vicinity of gold nanoparticles has been obtained using N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS), chemistry between amino groups of PANI and COOH groups of GOx. These AuNPs-PANI/ ITO and GOx/AuNPs-PANI/ITO composite films have been characterized using Fourier transform infra red (FTIR) and cyclic voltammetry (CV) techniques, respectively. The fast electron transfer from the modified PANI surface to electrode is indicated by the observed increase in amperometric response current of these GOx/AuNPs-PANI/ITO bioelectrodes. These GOx/AuNPs-PANI/ITO bioelectrodes exhibit response time of 10 s, linearity from 50 to 300 mg/dl and show value of apparent Michaelis-Menten constant (Km(app)) of 2.2 mM.     

Carbon nanotube-chitosan system for electrochemical sensing based on dehydrogenase enzymes

Multiwalled carbon nanotubes (CNT) were solubilized in aqueous solutions of a biopolymer chitosan (CHIT). The CHIT-induced solubilization of CNT facilitated their manipulations, including the modification of electrode surfaces for sensor and biosensor development. The colloidal solutions of CNT-CHIT were placed on the surface of glassy carbon (GC) electrodes to form robust CNT-CHIT films, which facilitated the electrooxidation of NADH. The GC/CNT-CHIT sensor for NADH required approximately 0.3 V less overpotential than the GC electrode. The susceptibility of CHIT to chemical modifications was explored in order to covalently immobilize glucose dehydrogenase (GDH) in the CNT-CHIT films using glutaric dialdehyde (GDI). The stability and sensitivity of the GC/CNT-CHIT-GDI-GDH biosensor allowed for the interference-free determination of glucose in the physiological matrix (urine). In pH 7.40 phosphate buffer solutions, linear least-squares calibration plots over the range 5-300 microM glucose (10 points) had slopes 80 mA M(-1) cm(-2) and a correlation coefficient 0.996. The detection limit was 3 microM glucose (S/N = 3). The CNT-CHIT system represents a simple and functional approach to the integration of dehydrogenases and electrodes, which can provide analytical access to a large group of enzymes for wide range of bioelectrochemical applications including biosensors and biofuel cells.     

Nitric oxide-releasing sol-gel particle/polyurethane glucose biosensors

A hybrid sol-gel/polyurethane glucose biosensor that releases nitric oxide is developed and characterized. The biosensor consists of a platinum electrode coated with four polymeric membranes including the following: (1) sol-gel with immobilized glucose oxidase (GOx); (2) polyurethane to protect the enzyme; (3) NO donor-modified sol-gel particle-doped polyurethane; and (4) polyurethane. This configuration was developed due to the drastic reduction in sensitivity observed for NO donor-modified sol-gel film-based glucose sensors. For the hybrid sol-gel/polyurethane biosensor, sol-gel particles are first modified with the NO donor and then incorporated into a polyurethane layer that is coated onto the preimmobilized GOx electrode. In this manner, the GOx layer is not exposed to the harsh conditions necessary to impart NO release ability to the biosensor, and only a minimal decrease in sensitivity due to the NO release is observed. The glucose response of the NO-releasing glucose biosensor and its NO generation profiles are reported. In addition, the stability of the sol-gel particles in the supporting polyurethane membrane is discussed.     

Fabrication of a glucose biosensor based on citric acid assisted cobalt ferrite magnetic nanoparticles

A novel and practical glucose biosensor was fabricated with immobilization of Glucose oxidase (GOx) enzyme on the surface of citric acid (CA) assisted cobalt ferrite (CF) magnetic nanoparticles (MNPs). This innovative sensor was constructed with glassy carbon electrode which is represented as (GOx)/CA-CF/(GCE). An explicit high negative zeta potential value (-22.4 mV at pH 7.0) was observed on the surface of CA-CF MNPs. Our sensor works on the principle of detection of H2O2 which is produced by the enzymatic oxidation of glucose to gluconic acid. This sensor has tremendous potential for application in glucose biosensing due to the higher sensitivity 2.5 microA/cm2-mM and substantial increment of the anodic peak current from 0.2 microA to 10.5 microA.     

Multi-walled carbon nanotubes-based glucose biosensor prepared by a layer-by-layer technique

The loading of multi-walled carbon nanotubes (MWNTs) and glucose oxidase (GOx) in the alternate layers of a glucose biosensor was first optimized based on a layer-by-layer construction on the surface of a graphite disk electrode. With the increasing of MWNTs/GOx layers, the response current to glucose was changed regularly and the response current reached a maximum value when the number of MWNTs/GOx layers was 6. Owing to a good electrical conductivity, strong adsorption and excellent bioconsistency of MWNTs, the (MWNTs/GOx)₆ films-coated glucose biosensor had an excellent electrochemical properties. The response current of the (MWNTs/GOx)₆ films-coated biosensor to 3 × 10^− 2 M glucose was 1.63 μA while the response time was only 6.7 s. The linear range and the lowest detectable concentration of this biosensor was 5 × 10^− 4∼1.5 × 10^− 2 M and 0.9 × 10^− 4 M, respectively.     

Deposition of platinum nanoparticles on organic functionalized carbon nanotubes grown in situ on carbon paper for fuel cells

Deposition of small Pt nanoparticles of the order of 2-2.5?nm on carbon nanotubes (CNTs) grown directly on carbon paper is demonstrated in this work. Sulfonic acid functionalization of CNTs is used as a means to facilitate the uniform deposition of Pt on the CNT surface. The organic molecules attached covalently to the CNT surface via electrochemical reduction of corresponding diazonium salts are treated with concentrated sulfuric acid and the sulfonic acid sites thus attached are used as molecular sites for Pt ion adsorption, which are subsequently reduced to yield the small Pt nanoparticles. Cyclic voltammograms reveal that, after removal of the organic groups during high temperature reduction, these Pt nanoparticles are in electrical contact with the carbon paper backing. A typical Pt loading of 0.09?mg?cm(-2) is achieved, that shows higher specific surface area of Pt than an E-TEK electrode with Pt loading of 0.075?mg?cm(-2). A membrane and electrode assembly (MEA) is prepared with a Pt/CNT electrode as cathode and an E-TEK electrode as anode, and it offers better performance than a conventional E-TEK MEA.     

Mediatorless,Reversible Optical Nanosensor Enabled through Enzymatic Pocket Doping

Single‐walled carbon nanotubes (SWCNTs) exhibit intrinsic near‐infrared fluorescence that benefits from indefinite photostability and tissue transparency, offering a promising basis for in vivo biosensing. Existing SWCNT optical sensors that rely on charge transfer for signal transduction often require exogenous mediators that compromise the stability and biocompatibility of the sensors. This study presents a reversible, mediatorless, near‐infrared glucose sensor based on glucose oxidase‐wrapped SWCNTs (GOx‐SWCNTs). GOx‐SWCNTs undergo a selective fluorescence increase in the presence of aldohexoses, with the strongest response toward glucose. When incorporated into a custom‐built membrane device, the sensor demonstrates a monotonic increase in initial response rates with increasing glucose concentrations between 3 × 10^?3 and 30 × 10^?3m and an apparent Michaelis–Menten constant of K_M(app) ≈ 13.9 × 10^?3m . A combination of fluorescence, absorption, and Raman spectroscopy measurements suggests a fluorescence enhancement mechanism based on localized enzymatic doping of SWCNT defect sites that does not rely on added mediators. Removal of glucose reverses the doping effects, resulting in full recovery of the fluorescence intensity. The cyclic addition and removal of glucose is shown to successively enhance and recover fluorescence, demonstrating reversibility that serves as a prerequisite for continuous glucose monitoring.     

Metallic nanoparticle-carbon nanotube composites for electrochemical determination of explosive nitroaromatic compounds

Metal nanoparticles (Pt, Au, or Cu) together with multiwalled and single-walled carbon nanotubes (MWCNT and SWCNT) solubilized in Nafion have been used to form nanocomposites for electrochemical detection of trinitrotoluene (TNT) and several other nitroaromatics. Electrochemical and surface characterization by cyclic voltammetry, AFM, TEM, SEM, and Raman spectroscopy confirmed the presence of metal nanoparticles on CNTs. Among various combinations tested, the most synergistic signal effect was observed for the nanocomposite modified glassy carbon electrode (GC) containing Cu nanoparticles and SWCNT solubilized in Nafion. This combination provided the best sensitivity for detecting TNT and other nitroaromatic compounds. Adsorptive stripping voltammetry for TNT resulted in a detection limit of 1 ppb, with linearity up to 3 orders of magnitude. Selectivity toward the number and position of the nitro groups in different nitroaromatics was very reproducible and distinct. Reproducibility of the TNT signal was within 7% (n = 8) from one electrode preparation to another, and the response signal was stable (+/-3.8% at 95% confidence interval) for 40 repeated analyses with 10 min of preconditioning. The Cu-SWCNT-modified GC electrode was demonstrated for analysis of TNT in tap water, river water, and contaminated soil.     

Platinum nanoparticle modified polyaniline-functionalized boron nitride nanotubes for amperometric glucose enzyme biosensor

A novel amperometric biosensor based on the BNNTs-Pani-Pt hybrids with Pt nanoparticle homogeneously decorated on polyaniline (Pani)-wrapped boron nitride nanotubes (BNNTs), was developed. It is shown that π interactions take place between BNNTs and polyaniline (Pani) located at N atoms from BNNTs and C atoms from Pani, resulting in the water solubility for the Pani wrapped BNNTs hybrids. The developed glucose biosensor displayed high sensitivity and stability, good reproducibility, anti-interference ability, especially excellent acid stability and heat resistance. The resulted BNNTs-Pani-Pt hybrid amperometric glucose biosensor exhibited a fast response time (within 3 s) and a linear calibration range from 0.01 to 5.5 mM with a high sensitivity and low detection limit of 19.02 mA M(-1) cm(-2) and 0.18 μM glucose (S/N = 3). Surprisedly, the relative activity of the GC/BNNTs-Pani-Pt-GOD electrode keeps almost no change in a range from pH 3 to 7. Futhermore, the BNNTs-Pani-Pt hybrid biosensor maintains a high GOD enzymatic activity even at a relatively high temperature of 60 °C. This might be attributed to the effect of electrostatic field and hydrophobia of BNNTs. The unique acid stability and heat resistance of this sensor indicate great promising application in numerous industrial and biotechnological operations involving harsh conditions.     

Electrochemical biosensor based on multi-walled carbon nanotubes and Au nanoparticles synthesized in chitosan

A new biosensor is prepared by cross-linking glucose oxidase (GOD) with glutaradehyde at the electrode combining Au nanoparticles (AuNP) with multi-walled carbon nanotubes (MWCNTs). Au nanoparticles-doped chitosan (CS) solution (AuNP-CS) is prepared by treating the CS solution followed by chemical reduction of Au (III) with NaBH4. MWCNTs are then dispersed in AuNP-CS solution. TEM, FT-IR, and UV-Vis show that the AuNP-CS solution is highly dispersed and stable. The synergistic effect between AuNP and CNTs of the AuNP-CNTs-CS material has been investigated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and amperometric methods. The modified glassy carbon electrode (GCE) allows low-potential detection of H2O2 with high sensitivity and fast response time. With the immobilization of GOD, a biosensor has been constructed. In phosphate buffer solutions (PBS, pH 7.0), nearly free interference determination of glucose has been realized at 0.4 V(vs. Ag/AgCl/3.0 M KCI) with a wide linear range from 2.0 x 10(-5) to 1.5 x 10(-2) M and a fast response time within 5s. The biosensor has been used to determine glucose in human serum samples and the results are satisfactory.     

Investigation of carbon nanofibers as support for bioactive substances

In this paper we have studied the adsorption properties of various bio-active systems onto the surface of carbon nanofibers (CNF) synthesized by chemical vapor deposition (CVD). Amino acids (alanine, aspartic acid, glutamic acid) and glucose oxidase (GOx) were adsorbed on CNF and the results were compared with those obtained when activated carbon (AC) was used as support. CNF and AC properties (hydrophilic or hydrophobic properties) were characterized by the pH value, the concentration of acidic/basic sites and by naphthalene adsorption. CNF with immobilized GOx was additionally investigated as a highly sensitive glucose biosensor. An amperometric method was used in an original manner to detect the changes in the specific activity of GOx, immobilized longer time on CNF. The method demonstrates that not the whole enzyme adsorbed onto CNF can catalyze the oxidation of glucose from the solution.