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Huangxian Ju

Publications and source records attributed to Huangxian Ju.

At least 19 recordsLinked to original sources

Detection of NADH and ethanol based on catalytic activity of soluble carbon nanofiber with low overpotential.

The excellent catalytic activity of a novel carbon nanomaterial, soluble carbon nanofiber (CNF) with good dispersion and wettability, to the oxidation of dihydronicotinamide adenine dinucleotide (NADH) was described for biosensing application. The edge plane sites and oxygen-rich groups presented on the CNF surface could be partially responsible for its electrocatalytic behavior, which induced a substantial decrease by 573 mV in the overpotential of NADH oxidation reaction (compared to a bare electrode). The CNF-coated electrode thus allowed highly sensitive amperometric detection of NADH with a low limit of detection (0.11 muM), low applied potential (+0.06 V), and minimization of surface fouling. Such ability of CNF to promote the electron transfer between NADH and the electrode suggested a new, promising biocompatible platform for development of dehydrogenase-based amperometric biosensors. With alcohol dehydrogenase (ADH) as a model, the ADH/CNF-modified electrode could be constructed by a simple casting process. The proposed biosensor showed rapid and highly sensitive amperometric response to ethanol with acceptable preparation reproducibility and excellent stability.

Alcohol Dehydrogenase↗

Sensitive reagentless electrochemical immunosensor based on an ormosil sol-gel membrane for human chorionic gonadotrophin.

A new organically modified silicate (ormosil) material was synthesized as a matrix to encapsulate enzyme labeled antibody for preparation of immunosensors. The ormosil matrix was prepared by hydrolyzing tetraethyoxysilane and (3-aminopropyl)triethoxysilane in weak alkali solution. It possessed three-dimensional ordered nanoporous structure with high electrical conductivity and good mechanical stability. Its hydrophilicity provided a microenvironment for retaining the biological activity of the immobilized protein. Particularly, using horseradish peroxidase-labeled human chorionic gonadotrophin antibody (HRP-anti-hCG) as a model, the immobilized HRP showed direct electron transfer at about -35 mV with a rate constant of 15.8+/-3.8s(-1). By a simple one-step immunoreaction between human serum chorionic gonadotrophin (hCG) in sample solution and the immobilized HRP-anti-hCG, the differential pulse voltammetric peak current of HRP decreased linearly with an increasing hCG concentration from 0.5 to 50 mIU/ml with a relatively low limit of detection of 0.3 mIU/ml at 3sigma. Excellent analytical performance, fabrication reproducibility and operational stability of the proposed biosensor indicated its promising application in clinical diagnostics.

Biosensing Techniques↗

Zirconia nanoparticles enhanced grafted collagen tri-helix scaffold for unmediated biosensing of hydrogen peroxide.

A novel, biocompatible, thermally steady, and nontoxic zirconia enhanced grafted collagen tri-helix scaffold was prepared on a graphite electrode. This scaffold provided a microenvironment for loading biomolecules and helped to retain their natural structure. UV-vis spectroscopy and scanning electron microscopy were used to characterize the scaffold and the structure of immobilized biomolecules. Using horseradish peroxidase (HRP) as an example, this scaffold accelerated its electron transfer and led to its direct electrochemical behavior with a good thermal stability up to 80 degrees C. The surface electron-transfer rate constant of the immobilized HRP was (5.55 +/- 0.43) s(-)(1) in 0.1 M pH 7.0 PBS at 18 degrees C. The immobilized HRP showed an electrocatalytic activity to the reduction of hydrogen peroxide (H(2)O(2)) without aid of an electron mediator. The linear response range of the biosensor for H(2)O(2) was from 1.0 to 73.0 microM with a correlation coefficient of 0.999 (n = 14), a limit of detection down to 0.25 microM and an apparent Michaelis-Menten constant of (0.28 +/- 0.02) mM. The biosensor exhibited high sensitivity, acceptable stability, and reproducibility. The ZrO(2) grafted collagen provided an excellent matrix for protein immobilization and biosensor preparation.

Biosensing Techniques↗

Reagentless biosensor for hydrogen peroxide based on immobilization of protein in zirconia nanoparticles enhanced grafted collagen matrix.

A novel matrix, zirconia nanoparticles enhanced grafted collagen (ZrO2-grafted collagen) hybrid composite, for immobilization of protein and biosensing was developed. The scanning electron microscopy, UV-vis and Fourier transform infrared spectra, and electrochemical measurements showed that the matrix was well biocompatible and could retain the bioactivity of immobilized protein to a large extent. The direct electron transfer of the immobilized myoglobin (Mb) exhibited a couple of stable and well-defined redox peaks with the formal potential of -336 mV (versus SCE) in 0.1M pH 7.0 PBS. This matrix could accelerate the electron transfer between Mb and the electrode with a surface-controlled process and an electron transfer rate constant of 3.58+/-0.35s-1 at 10-500 mVs-1. The Mb immobilized in the matrix showed a high thermal stability up to 70 degrees C and an electrocatalytic activity to the reduction of hydrogen peroxide (H2O2) without the help of an electron mediator. The linear response range of the biosensor to H2O2 concentration was from 1.0 to 85.0 microM with the limit of detection of 0.63 microM at a signal-to-noise ratio of 3sigma. The biosensor exhibited high sensitivity, acceptable stability and reproducibility. This work opened a way for the further study on the direct electron transfer and biosensing application of the immobilized protein in collagen-related matrices.

Biosensing Techniques↗

Flow-through multianalyte chemiluminescent immunosensing system with designed substrate zone-resolved technique for sequential detection of tumor markers.

A novel flow-through immunosensing system for performing a multianalyte chemiluminescent determination in a single run was designed. A new analytical strategy of substrate zone-resolved technique was proposed. Using carcinoma antigen 125 (CA 125) and carcinoembryonic antigen (CEA) as model analytes, the capture antibodies for CA 125 and CEA were immobilized on an UltraBind aldehyde-activated membrane to act as an immunoreactor, to which the mixture of CA 125, CEA, and their corresponding tracers, horseradish peroxidase (HRP)-labeled anti-CA 125 and alkaline phosphatase (ALP)-labeled anti-CEA, was introduced for on-line incubation. The substrates for HRP and ALP were then delivered into the detection cell sequentially to perform substrate zone-resolved immunoassay by a sandwich format. Under optimal conditions, CA 125 and CEA could be assayed in the ranges of 5.0-100 units/mL and 1.0-120 ng/mL, respectively. The whole assay process including incubation, wash, detection, and regeneration could be completed in 35 min. The serum samples from the clinic were assayed with the proposed method, and the results were in acceptable agreement with the reference values. This method and the strategy of substrate zone-resolved technique could be further developed for high-throughput multianalyte immunoassay.

Biomarkers, Tumor↗

Layer-by-layer hydroxymethyl ferrocene modified sensor for one-step flow/stop-flow injection amperometric immunoassay of alpha-fetoprotein.

A rapid one-step flow/stop-flow injection amperometric immunoassay for alpha-fetoprotein (AFP) using a novel home-produced electrochemical sensor was proposed. The sensor was prepared using layer-by-layer adsorption of positively charged poly(allylamine) (PAA) and negatively charged hydroxymethyl ferrocene on a screen-printed electrode (SPE). The electrochemistry of the immobilized ferrocene moieties showed a surface-controlled electrode process. Based on an electrochemical enzyme-linked immunoassay with the immobilized ferrocene moieties as an electron transfer mediator between the electrode and the horseradish peroxidase (HRP)-labeled anti-AFP antibody, a calibration curve with two linear ranges from 5 to 20 and 20 to 150 ng ml-1 and a detection limit of 2 ng ml-1 for AFP determination was obtained under the optimized conditions of 0.891 ml min-1 flow rate, 20 microl injection volume and +25 mV applied potential. The sensor showed good repeatability and reproducibility and retained more than 95% of its original signal after 15 days of storage. The proposed method eliminated the need for washing and addition of any substrate or mediator. The complete assay could be handled in less than 25 min with a one-step injection of a 40 microl sample solution. The proposed method would be valuable for the diagnosis and monitoring of carcinoma and its metastasis.

Biosensing Techniques↗

Electrogenerated chemiluminescence detection of amino acids based on precolumn derivatization coupled with capillary electrophoresis separation.

A novel method for highly sensitive detection of primary and secondary amino acids with selective derivatization using acetaldehyde as a new derivatization reagent was proposed by capillary electrophoresis (CE) coupled with electrogenerated chemiluminescence (ECL) of tris(2,2'-bipyridine)ruthenium(II). The precolumn derivatization of these amino acids with acetaldehyde was performed in aqueous solution at room temperature for 1 h. Upon optimized derivatization, the ECL intensities and detection sensitivities of the amino acids were significantly enhanced by 20-70 times. Using four amino acids, arginine, proline, valine, and leucine, as model compounds, their derivatives could be completely separated by CE and sensitively detected by ECL within 22 min. The linear ranges were 0.5-100 microM for arginine and proline and 5-1000 microM for valine and leucine with the detection limits of 1 x 10(-7) (0.5 fmol, arginine), 8 x 10(-8) (0.4 fmol, proline), 1 x 10(-6) (5 fmol, valine), and 1.6 x 10(-6) M (8 fmol, leucine) at a signal-to-noise ratio of 3. The derivatization reactions and ECL process of amino acids were also proposed based on in situ Fourier transform infrared and ultraviolet spectrometric analyses.

2,2'-Dipyridyl↗

Simultaneous electrochemiluminescence determination of sulpiride and tiapride by capillary electrophoresis with cyclodextrin additives.

Sulpiride and tiapride are often used in the treatment of depression, schizophrenia and psychopathology of senescence, gastric or duodenal ulcers and are also partly excreted by kidney. This work developed a simple and sensitive method for their simultaneous monitoring in human urine based on capillary electrophoresis coupled with electrochemiluminescence detection by end-column mode. beta-Cyclodextrin (beta-CD) was used as an additive to the running buffer to obtain the absolute separation of sulpiride and tiapride. Under optimized conditions the proposed method displayed a linear range from 1.0 x 10(-7) to 1.0 x 10(-4) M for both sulpiride and tiapride with the correlation coefficients more than 0.995 (n = 6). Their limits of detection were 1.0 x 10(-8) M (45 amol) and 1.5 x 10(-8) M (68 amol) at a signal to noise ratio of 3, respectively. The relative standard deviations for six determinations of 2.0 microM sulpiride and 3.0 microM tiapride were 1.8 and 2.5%, respectively. For practical application an extract step with ethyl acetate at pH 11 was performed to eliminate the influence of ionic strength in sample. The recoveries of sulpiride and tiapride at different levels in human urine were between 84 and 95%, which showed that the method was valuable in clinical and biochemical laboratories for monitoring sulpiride and tiapride for various purposes.

Electrochemistry↗

Flow-injection chemiluminescent immunoassay for alpha-fetoprotein based on epoxysilane modified glass microbeads.

A flow-injection chemiluminescent immunoassay system based on a novel transparent immunoaffinity reactor is proposed for the quantitation of alpha-fetoprotein. The reactor prepared with alpha-fetoprotein immobilized epoxysilane modified glass microbeads was used as an immunosensor for chemiluminescent detection. With a non-competitive immunoassay format, the proposed immunosensor system is a low cost, flexible and rapid assay for alpha-fetoprotein. After an off-line incubation of the analyte alpha-fetoprotein with horseradish peroxidase-labeled alpha-fetoprotein antibody as enzyme tracer, the mixture was injected into the reactor, which led to trapping of the free enzyme tracer by the reactor. The trapped enzyme tracer was detected by the p-iodophenol-luminol-H2O2 chemiluminescence system. Under optimal conditions, the decrease in chemiluminescence intensity was proportional to the alpha-fetoprotein concentration in the range of 5.0-100 ng/ml with a detection limit of 2.7 ng/ml at a signal/noise ratio of 3. The immunosensor system showed an acceptable reproducibility and stability. Clinical serum samples were assayed with this method and the results were in acceptable agreement with those obtained from immunoradiometric assay.

Biosensing Techniques↗

Binding of acetylcholinesterase to multiwall carbon nanotube-cross-linked chitosan composite for flow-injection amperometric detection of an organophosphorous insecticide.

A novel method for immobilization of acetylcholinesterase (AChE) by binding covalently to a cross-linked chitosan-multiwall carbon nanotube (MWNT) composite is described. In addition a sensitive, fast, cheap and automatizable flow injection detection of an organophosphorous insecticide was developed. The MWNTs were homogeneously distributed in the chitosan membrane which showed a homogeneous porous structure. The immobilized AChE could catalyze the hydrolysis of acetylthiocholine with a K(M)app value of 177 microM to form thiocholine, which was then oxidized to produce detectable signal in a linear range of 1.0-500 microM and fast response. MWNTs could catalyze the electrooxidation of thiocholine, thus increasing detection sensitivity. Based on the inhibition of an organophosphorous insecticide on the enzymatic activity of AChE, using Sulfotep as a model compound, the conditions for the flow-injection detection of the insecticide were optimized. Both biocompatibility of chitosan and inherent conductive properties of MWNTs favored the detection of the insecticide from 1.5 to 80 microM along with good stability and reproducibility. 95 % reactivation from inhibited AChE could be regenerated by using 2-pyridinealdoxime methiodide within 15 min for 15 times. The detection of Sulfotep samples exhibited satisfactory results. The proposed flow-injection analysis device can be applied to automated determination and characterization of enzyme inhibitors.

Acetylcholinesterase↗

A disposable electrochemical immunosensor for flow injection immunoassay of carcinoembryonic antigen.

A new simple immunoassay method for carcinoembryonic antigen (CEA) detection using a disposable immunosensor coupled with a flow injection system was developed. The immunosensor was prepared by coating CEA/colloid Au/chitosan membrane at a screen-printed carbon electrode (SPCE). Using a competitive immunoassay format, the immunosensor inserted in the flow system with an injection of sample and horseradish peroxidase (HRP)-labeled CEA antibody was used to trap the labeled antibody at room temperature for 35 min. The current response obtained from the labeled HRP to thionine-H(2)O(2) system decreased proportionally to the CEA concentration in the range of 0.50-25 ng/ml with a correlation coefficient of 0.9981 and a detection limit of 0.22 ng/ml (S/N=3). The immunoassay system could automatically control the incubation, washing and current measurement steps with good stability and acceptable accuracy. Thus, the proposed method proved its potential use in clinical immunoassay of CEA.

Biosensing Techniques↗

Simultaneous determination of ethamsylate, tramadol and lidocaine in human urine by capillary electrophoresis with electrochemiluminescence detection.

Ethamsylate, tramadol and lidocaine, partly excreted by the kidney, are generally used as hemostatic, analgesic and local anesthetic in surgery. We developed a simple and sensitive method for their simultaneous monitoring in human urine based on CE coupled with electrochemiluminescence detection by end-column mode. Under optimized conditions the proposed method yielded linear ranges from 5.0 x 10(-8) to 5.0 x 10(-5), 1.0 x 10(-7) to 1.0 x 10(-4) and 1.0 x 10(-7) to 1.0 x 10(-4) M with LODs of 8.0 x 10(-9) M (36 amol), 1.6 x 10(-8) M (72 amol) and 1.0 x 10(-8) M (45 amol) (S/N = 3) for ethamsylate, tramadol and lidocaine, respectively. The RSD for their simultaneous detection at 1.0 x 10(-6) M was 2.1, 2.8 and 3.2% (n = 7), respectively. For practical application an extraction step with ethyl acetate at pH 11 was performed to eliminate the influence of the sample ionic strength. The recoveries of ethamsylate, tramadol and lidocaine at different levels in human urine were between 87 and 95%. This method was used for simultaneous detection of ethamsylate, tramadol and lidocaine in clinic urine samples from two medicated patients. It was valuable in clinical and biochemical laboratories for monitoring these drugs for various purposes.

Electrophoresis, Capillary↗

[Extraction and determination of short-chain fatty acids in biological samples].

Short-chain fatty acids are organic acids with 1 - 6 carbon atoms. Their physiological functions and clinical applications have attracted considerable attention. Their high polarity, low ultraviolet (UV) absorbance, low contents in biological samples, high volatility, good water-solubility and easy adsorption on metal and glass surfaces result in some difficulties in their separation and determination. This review focuses on the methods for extraction of short-chain fatty acids in biological samples such as feces, urine, blood and culture solutions by distillation, high speed centrifugation, liquid-solid extraction, solid phase microextration, supercritical fluid extraction and liquid phase microextraction and determination by gas chromatography, high performance liquid chromatography and capillary electrophoresis. Sixty-three references are cited.

Adsorption↗

Multilayer membranes for glucose biosensing via layer-by-layer assembly of multiwall carbon nanotubes and glucose oxidase.

A bilayer of the polyelectrolytes poly(dimethyldiallylammonium chloride) (PDDA) and poly(sodium 4-styrenesulfonate) (PSS) was formed on a 3-mercapto-1-propanesulfonic-acid-modified Au electrode. Subsequently, multiwall carbon nanotubes (MWCNTs) wrapped by positively charged PDDA were assembled layer-by-layer with negatively charged glucose oxidase (GOx) onto the PSS-terminated bilayer. Electrochemical impedance spectroscopy and atomic force microscopy were adopted to monitor the regular growth of the PDDA-MWCNTs/GOx bilayers. Using GOx as a model enzyme, the assembled multilayer membranes showed some striking features such as the adsorbed form of GOx on individual MWCNT, uniformity, good stability, and electrocatalytic activity toward oxygen reduction. Based on the consumption of dissolved oxygen during the oxidation process of glucose catalyzed by the immobilized GOx, a sensitive amperometric biosensor was developed for the detection of glucose up to 5.0 mM with a detection limit of 58 microM. The sensitivity increased with increasing sensing layers up to five bilayers. Ascorbic acid and uric acid did not cause any interference due to the use of a low operating potential. The present method showed high reproducibility for the fabrication of carbon-nanotubes-based amperometric biosensors.

Biosensing Techniques↗

A gold nanoparticles/sol-gel composite architecture for encapsulation of immunoconjugate for reagentless electrochemical immunoassay.

A highly hydrophilic, non-toxic and conductive colloidal gold nanoparticle/titania sol-gel composite membrane with a low contact angle was prepared on a glassy carbon electrode via a vapor deposition method. With human chorionic gonadotrophin (hCG) as a model antigen and encapsulation of horseradish peroxidase-labeled hCG antibody (HRP-anti-hCG) in the composite architecture, this membrane could be used for reagentless electrochemical immunoassay. It displayed a porous and homogeneous composite architecture without the aggregation of the immobilized protein molecules. The presence of gold nanoparticles provided a congenial microenvironment for adsorbed biomolecules and decreased the electron transfer impedance, leading to a direct electrochemical behavior of the immobilized HRP. The formation of immunoconjugate by a simple one-step immunoreaction between hCG in sample solution and the immobilized HRP-anti-hCG introduced a barrier of direct electrical communication between the immobilized HRP and the electrode surface. Under optimal conditions, the hCG analyte could be determined in two linear ranges from 0.5 to 5.0 mIU/mL and 5.0 to 30 mIU/mL with a relatively low detection limit of 0.3 mIU/mL at 3sigma. The hCG immunosensor exhibited good precision, high sensitivity, acceptable stability, accuracy and reproducibility. This composite membrane could be used efficiently for the entrapment of different biomarkers and clinical applications.

Biomarkers↗

Rapid detection of ssDNA and RNA using multi-walled carbon nanotubes modified screen-printed carbon electrode.

A method for rapid sensitive detection of DNA or RNA was designed using a composite screen-printed carbon electrode modified with multi-walled carbon nanotubes (MWNTs). MWNTs showed catalytic characteristics for the direct electrochemical oxidation of guanine or adenine residues of signal strand DNA (ssDNA) and adenine residues of RNA, leading to indicator-free detection of ssDNA and RNA concentrations. With an accumulation time of 5 min, the proposed method could be used for detection of calf thymus ssDNA ranging from 17.0 to 345 microg ml(-1) with a detection limit of 2.0 microg ml(-1) at 3 sigma and yeast tRNA ranging from 8.2 microg ml(-1) to 4.1 mg ml(-1). AC impedance was employed to characterize the surface of modified electrodes. The advantages of convenient fabrication, low-cost detection, short analysis time and combination with nanotechnology for increasing the sensitivity made the subject worthy of special emphasis in the research programs and sources of new commercial products.

Biocompatible Materials↗

Electrochemical immunoassay of membrane P-glycoprotein by immobilization of cells on gold nanoparticles modified on a methoxysilyl-terminated butyrylchitosan matrix.

A strategy to detect P-glycoprotein (P-gp) on cell membrane and quantify the cell number using electrochemical immunoassay was developed by effective surface immunoreactions and immobilization of cells on a highly hydrophilic interface, which was constructed by adsorption of colloidal gold nanoparticles on a methoxysilyl-terminated (Mos) butyrylchitosan modified glassy carbon electrode (Au-CS/GCE). Atomic force microscopy studies proved that the nanoparticles adsorbed on Mos-butyrylchitosan were efficient in preventing the cell leakage and retaining the activity of immobilized living K562/ADM leukemic cells. The incubation with P-gp monoclonal antibody and then the secondary alkaline phosphatase (AP) conjugated antibody introduced AP onto the K562/ADM cell immobilized on Au-CS/GCE. The bound AP led to an amperometric response of 1-naphthyl phosphate. Under optimal conditions the response was proportional to the logarithm of cell concentration in the range from 5.0 x 10(4) to 1.0 x 10(7) cells mL(-)(1) with a detection limit of 1.0 x 10(4) cells mL(-)(1). The results were comparable to flow cytometric analysis of P-gp expression. This proposed method was practical, convenient, and significant in the clinic and cytobiology.

ATP Binding Cassette Transporter, Subfamily B, Mem↗