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Biomedical subjects

Jing-Juan Xu

Publications and source records attributed to Jing-Juan Xu.

At least 19 recordsLinked to original sources

Glucose microfluidic biosensors based on immobilizing glucose oxidase in poly(dimethylsiloxane) electrophoretic microchips.

Here we reported a novel microfluidic biosensor with an on-column immobilized enzyme microreactor. The fabrication approach of this biosensor is simple and the enzyme microreactors with controlled sizes can be placed at any desired position on the microchip. Taking glucose oxidase (GOx) as an example, electroosmotic flow (EOF) as a driving force and amperometry as a detection method, the performance of biosensors were modulated by changing the length of enzyme reactor from 0.5 cm to 3 cm, and the linear ranges were changed from 0-8.0 mM to 0-30.0 mM with the detection limits from 42 microM to 6.5 microM. The enzyme reactor remained its 65% activity after 23 days storage. It also showed good anti-interference ability and was used to quantify glucose in human serum samples.

Biosensing Techniques↗

Direct electron transfer and electrocatalysis of hemoglobin adsorbed on mesoporous carbon through layer-by-layer assembly.

Using chitosan as an effective linker between CMK-3 and glassy carbon electrode surface, {Hb/CMK-3}n multilayer film-modified electrodes were constructed through layer-by-layer assembly. The morphology of thus-formed {Hb/CMK-3}n film was characterized by scanning electron micrographs, and the interaction of hemoglobin (Hb) with CMK-3 was studied by UV-vis spectroscopy and electrochemical methods. Under optimal conditions, {Hb/CMK-3}6 film showed a couple of stable and well-defined redox peaks at about -377 and -296 mV in pH 7.0 buffers. Furthermore, the {Hb/CMK-3}6 film displayed excellent electrocatalysis to the reduction of both H2O2 and O2. Based on thus-formed film and its direct electron transfer behavior, a novel biosensor was presented for the determination of H2O2 ranging from 1.2 to 57 muM with the detection limit of 0.6microM at S/N=3. CMK-3 provided a desirable matrix for protein immobilization and biosensor preparation.

Adsorption↗

Novel coupling mechanism-based imaging approach to scanning electrochemical microscopy for probing the electric field distribution at the microchannel end.

A novel coupling mechanism-based imaging approach to scanning electrochemical microscopy (SECM) was used to image the distribution of electric field at the end channel of a poly(dimethylsiloxane) (PDMS) capillary electrophoresis (CE) microchip in the absence of redox species. The coupling imaging mechanism was systematically investigated and qualitatively illustrated. It was proved that the distribution of solution potentials within the scanning plane caused a different reduction rate of water at the tip electrode, which led to the variation in tip current. Within the scanning plane, the solution potentials measured in the central area of the microchannel were usually higher than those measured outside. The SECM images showed a strong dependence on tip potential, tip-to-channel distance, and separation potential. According to the Tafel equation, SECM images were converted to parameters that directly showed the distribution of solution potential. Change in the solution potential along the central axial line of the microchannel was also continuously sensed by allowing the tip to approach the microchannel in the presence of high voltage. Using dopamine as a model compound, the effect of solution potential on electrochemical detection was estimated by detecting separation parameters.

Dimethylpolysiloxanes↗

Determination of morphine and codeine in urine using poly(dimethylsiloxane) microchip electrophoresis with electrochemical detection.

In this paper, a poly(dimethylsiloxane) (PDMS) microchip with electrochemical (EC) detection was developed for rapid separation and detection of morphine and codeine. It was found that morphine and codeine were well separated within 140 s in phosphate buffer solution (PBS) (pH 6.6, 40 mM)-beta-cyclodextrin (beta-CD) (20 mM)-acetonitrile (30%, v/v). The detection limit was 0.2 microM for morphine and 1 microM for codeine. The protocol was successfully applied to monitoring the amount of morphine and codeine in human urine. Compared with the conventional methods, the presented method had many advantages such as lower instrument cost, less reagent consumption and shorter analysis time.

Acetonitriles↗

A simple method to fabricate a chitosan-gold nanoparticles film and its application in glucose biosensor.

A novel film of chitosan-gold nanoparticles is fabricated by a direct and facile electrochemical deposition method and its application in glucose biosensor is investigated. HAuCl(4) solution is mixed with chitosan and electrochemically reduced to gold nanoparticles, which can be stabilized by chitosan and electrodeposited onto glassy carbon electrode surfaces along with the electrodeposition of chitosan. Then a model enzyme, glucose oxidase (GOD) is immobilized onto the resulting film to construct a glucose biosensor through self-assembly. The resulting modified electrode surfaces are characterized with both AFM and cyclic voltammetry. Effects of chitosan and HAuCl(4) concentration in the mixture together with the deposition time and the applied voltage on the amperometric response of the biosensor are also investigated. The linear range of the glucose biosensor is from 5.0 x 10(-5) approximately 1.30 x 10(-3) M with a Michaelis-Menten constant of 3.5 mM and a detection limit of about 13 microM.

Biosensing Techniques↗

Nanocrystalline diamond modified gold electrode for glucose biosensing.

Boron-doped diamond has drawn much attention in electrochemical sensors. However there are few reports on non-doped diamond because of its weak conductivity. Here, we reported a glucose biosensor based on electrochemical pretreatment of non-doped nanocrystalline diamond (N-NCD) modified gold electrode for the selective detection of glucose. N-NCD was coated on gold electrode and glucose oxidase (GOx) was immobilized onto the surfaces of N-NCD by forming amide linkages between enzyme amine residues and carboxylic acid groups on N-NCD. The anodic pretreatment of N-NCD modified electrode not only promoted the electron transfer rate in the N-NCD thin film, but also resulted in a dramatic improvement in the reduction of the dissolved oxygen. This performance could be used to detect glucose at negative potential through monitoring the current change of oxygen reduction. The biosensor effectively performs a selective electrochemical analysis of glucose in the presence of common interferents, such as ascorbic acid (AA), acetaminophen (AP) and uric acid (UA). A wide linear calibration range from 10 microM to 15 mM and a low detection limit of 5 microM were achieved for the detection of glucose.

Biosensing Techniques↗

Off-line form of the Michaelis-Menten equation for studying the reaction kinetics in a polymer microchip integrated with enzyme microreactor.

We firstly transformed the traditional Michaelis-Menten equation into an off-line form which can be used for evaluating the Michaelis-Menten constant after the enzymatic reaction. For experimental estimation of the kinetics of enzymatic reactions, we have developed a facile and effective method by integrating an enzyme microreactor into direct-printing polymer microchips. Strong nonspecific adsorption of proteins was utilized to effectively immobilize enzymes onto the microchannel wall, forming the integrated on-column enzyme microreactor in a microchip. The properties of the integrated enzyme microreactor were evaluated by using the enzymatic reaction of glucose oxidase (GOx) with its substrate glucose as a model system. The reaction product, hydrogen peroxide, was electrochemically (EC) analyzed using a Pt microelectrode. The data for enzyme kinetics using our off-line form of the Michaelis-Menten equation was obtained (K(m) = 2.64 mM), which is much smaller than that reported in solution (K(m) = 6.0 mM). Due to the hydrophobic property and the native mesoscopic structure of the poly(ethylene terephthalate) film, the immobilized enzyme in the microreactor shows good stability and bioactivity under the flowing conditions.

Enzyme Stability↗

Proteins modification of poly(dimethylsiloxane) microfluidic channels for the enhanced microchip electrophoresis.

This report described proteins modification of poly(dimethylsiloxane) (PDMS) microfluidic chip based on layer-by-layer (LBL) assembly technique for enhancing separation efficiency. Two kinds of protein-coated films were prepared. One was obtained by successively immobilizing the cationic polyelectrolyte (chitosan, Chit), gold nanoparticles (GNPs), and protein (albumin, Albu) to the PDMS microfluidic channels surface. The other was achieved by sequentially coating lysozyme (Lys) and Albu. Neurotransmitters (dopamine, DA; epinephrine, EP) and environmental pollutants (p-phenylenediamine, p-PDA; 4-aminophenol, 4-AP; hydroquinone, HQ) as two groups of separation models were studied to evaluate the effect of the functional PDMS microfluidic chips. The results clearly showed these analytes were efficiently separated within 140 s in a 3.7 cm long separation channel and successfully detected with in-channel amperometric detection mode. Experimental parameters in two protocols were optimized in detail. The detection limits of DA, EP, p-PDA, 4-AP, and HQ were 2.0, 4.7, 8.1, 12.3, and 14.8 microM (S/N=3) on the Chit-GNPs-Albu coated PDMS/PDMS microchip, and 1.2, 2.7, 7.2, 9.8, and 12.2 microM (S/N=3) on the Lys-Albu coated one, respectively. In addition, through modification, the more homogenous channel surface displayed higher reproducibility and better stability.

Buffers↗

Patterning microbeads inside poly(dimethylsiloxane) microfluidic channels and its application for immobilized microfluidic enzyme reactors.

We propose a convenient and reliable approach for immobilizing microbeads on poly(dimethylsiloxane) (PDMS) microchips. It is built upon a simple fabrication procedure of PDMS chip through directly printing the master with an office laser printer which was described in our previous work (J. Chromatogr. A 2005, 1089, 270-275). On the printed toners used as the positive relief of the master, microbeads were immobilized by a thermal treatment and then transferred to the surface of the microchip by direct molding of the prepolymer on the master. With this approach, the region-selective immobilization of microbeads and the fabrication of PDMS microchips can be accomplished at the same time. Then, using these microbeads as supports, further modification with enzyme was achieved. Surface characteristics of the microbeads-modified PDMS microchannels were investigated with scanning electron microscope, atomic force microscope, and inverse fluorescence microscope. The electrokinetic properties of the native PDMS and the modified PDMS chips were also compared. Based on this approach, an immobilized glucose oxidase (GOD) reactor was constructed and the reaction using glucose as substrate was studied. All these experiments aim to show that the proposed approach may have a good potential in the study of biochemistry and other related areas.

Catalysis↗

Interfacing myoglobin to graphite electrode with an electrodeposited nanoporous ZnO film.

A biocompatible, nanoporous ZnO film was prepared on graphite electrode by the simple electrodeposition method. Based on the film's strong adsorption ability and friendly microenvironment, it can be used as a good matrix to immobilize myoglobin (Mb) through simple adsorption. Moreover, the entrapped Mb realized direct electron transfer with the electrode and displayed an elegant catalytic activity toward the reduction of hydrogen peroxide, nitrite, and trichloroacetic acid, by which the mediator-free biosensors could be fabricated. Atomic force microscopy, UV-Vis spectra, electrochemical impedance spectroscopy, and cyclic voltammetry, etc. were used to characterize the nanoporous ZnO film and Mb-modified ZnO film. Compared to other methods, electrodeposition of porous material supplied a more simple and convenient approach to prepare biocompatible materials for biosensors.

Adsorption↗

Electroosmotic flow in poly(dimethylsiloxane) microchannels.

This paper reports on the study of electroosmotic flow (EOF) in poly(dimethylsiloxane) (PDMS) microchannels on the basis of indirect amperometric detection method. Gradual increase of EOF rate in freshly prepared PDMS microchannels was observed with the running buffer of phosphate buffer solution (PBS). With the same concentration (10 mM) of PBS containing different cations and the same pH value (7.0) and, the time of the stable EOF in PDMS microchannels under the applied separation voltage of 1000 V was 49.8 s (Li+ -PBS), 57.1 s (Na+ -PBS), 91 s (K+ -PBS), respectively. Meanwhile, the different adsorption of cations (Li+, Na+ and K+) on hydrophobic PDMS wall was observed through their separation in PDMS microchannels. Such experimental results demonstrated that the EOF in PDMS microchannels came from the cations and anions adsorbed on PDMS wall. This study would not only help us understand the surface state of PDMS, but also provide a useful guidance for establishing the effective surface modification methods in PDMS microchip CE.

Dimethylpolysiloxanes↗

Simple method for the separation and detection of native amino acids and the identification of electroactive and non-electroactive analytes.

Detection of native amino acids was accomplished using a capillary electrophoresis (CE)-amperometric detection system, in which a single carbon fiber cylinder (CFC) working electrode instead of a carbon fiber disc (CFD) electrode was mounted in the end part of a poly(dimethylsiloxane) (PDMS)/glass hybrid microchannel. Similar to that using CFD working electrode, here, the electrochemical reduction reaction at the working electrode is also coupled from the separation high voltage system, the coupling degree is related to the in-channel length of the CFC. This property simplifies the fabrication procedure of the working electrode and also provides a convenient and sensitive means for the determination of non-electroactive ions by amperometry. The present detection mode is successfully used to electrochemically detect non-electroactive arginine (Arg), threonine (Thr), glutamic acid (Glu) and electroactive cysteine (Cys). Furthermore, by simply changing the detection potential, we can easily distinguish peak mobilities of electroactive amino acids from that of non-electroactive amino acids.

Amino Acids↗

Multilayer membranes via layer-by-layer deposition of organic polymer protected Prussian blue nanoparticles and glucose oxidase for glucose biosensing.

Polyelectrolyte multilayers (PEMs) are now widely used for bioanalytical applications. In this work, a bilayer of poly(diallydimethylammonium chloride) (PDDA) and poly(sodium 4-styrenesulfonate) (PSS) is consecutively adsorbed on 3-mercapto-1-propanesulfonic acid modified Au electrode surfaces, forming stable, ultrathin multilayer films. Subsequently, Prussian blue nanoparticles protected by PDDA (denoted as P-PB) and negatively charged glucose oxidase (GOx) are consecutively adsorbed onto the PSS-terminated bilayer. The growth of each of the P-PB/GOx bilayers is followed quantitatively using UV-visible absorption spectroscopy and the electrochemical method. The P-PB nanoparticles can catalyze the electroreduction of hydrogen peroxide formed from enzymatic reaction at lower potential and inhibit the responses of interferents, such as ascorbic acid (AA) and uric acid (UA). Performance of the multilayer films can be tailored by controlling the number of bilayers. Under optimal conditions, a linear range of 0.10 to 11.0 mM and a detection limit of 10 microM were achieved. The glucose biosensor has good stability and reproducibility.

Biosensing Techniques↗

Fabrication of poly(dimethylsiloxane) microfluidic system based on masters directly printed with an office laser printer.

Applications of poly(dimethylsiloxane) (PDMS)-based microfluidic systems are more popular nowadays. Previous fabrication methods of the masters for PDMS microchannels require complicated steps and/or special device. In this paper, we demonstrated that the toner printed on the transparency film with the office laser printer (1200 dpi) can be used as the positive relief of the masters. The transparency film was printed in two steps in order to obtain the same printing quality for the crossed lines. With the laser-printed master, the depth of the fabricated PDMS microchannels was ca. 10 microm and the smallest width was ca. 60 microm. Surface characteristics of the PDMS/PDMS microchannels were performed with SEM. Their electrokinetic properties were investigated by the aids of the measurement of electroosmotic flow (EOF) and the Ohm's curve. Using the PDMS/PDMS microchip CE systems, electroactive biological molecules and non-electroactive inorganic ions were well separated, respectively. This simple approach could make it easy to carry out the studies of PDMS microfluidic systems in more general labs without special devices.

Dimethylpolysiloxanes↗

Direct electrochemistry and electrocatalysis of heme proteins immobilized on gold nanoparticles stabilized by chitosan.

Three heme proteins, myoglobin, hemoglobin, and cytochrome c, have been adsorbed onto chitosan-stabilized gold nanoparticles (Chit-Aus) modified Au electrode via a molecule bridge like cysteine. UV-vis spectra indicated that the proteins on Chit-Aus films retained near-native secondary structures. The fabricated procedures and electrochemical behaviors of proteins on such an interface were characterized with electrochemical impedance spectra and cyclic voltammetric techniques. It was demonstrated that Chit-Aus film could not only offer a friendly environment to immobilize protein molecules but also enhance the electron transfer ability between protein molecules and underlying electrode. The effects of scan rate and pH on the electrochemical behaviors of each heme protein are discussed in detail. The resultant electrode displayed an excellent electrocatalytic response to the reduction of H(2)O(2), long-term stability, and good reproducibility.

Animals↗

Electrochemically deposited chitosan hydrogel for horseradish peroxidase immobilization through gold nanoparticles self-assembly.

A new strategy for immobilization of horseradish peroxidase (HRP) has been presented by self-assembling gold nanoparticles on chitosan hydrogel modified Au electrode. From a mildly acidic chitosan solution, a chitosan film is electrochemically deposited on Au electrode surface via a negative voltage bias. This process is accompanied by the hydrogen evolution reaction, and the released hydrogen gas made the deposited chitosan film with porous structure, which facilitates the assembly of gold nanoparticles and HRP. The resulting substrates were characterized by atomic force microscopy (AFM) and electrochemical impedance spectroscopy (EIS). The immobilized HRP displayed an excellent catalytic property to the reduction of H2O2 in the presence of methylene blue mediator. The resulting biosensor (HRP-modified electrode) showed a wide dynamic range of 8.0 microM-15 mM H2O2, and the linear ranges were 8.0 microM-0.12 mM and 0.50-12 mM, with a detection limit of 2.4 microM estimated at a signal-to-noise ratio of 3. Moreover, the biosensor remained about 85% of its original sensitivity after four weeks' storage.

Animals↗

Extended-range glucose biosensor via layer-by-layer assembly incorporating gold nanoparticles.

We report on a glucose oxidase (GOx)/polyelectrolyte (PE)/gold nanoparticle (AuNP) multilayer films that can be utilized as efficient glucose biosensors by layer-by-layer self-assembly method. Electrochemical impedance spectroscopy (EIS) and UV-visible spectroscopy were adopted to monitor the regular growth of the multilayer films. The role of gold nanoparticles integrated in the multilayer films not only increase the amount and activity of GOx, but also significantly improve the electron-transfer characteristics of the films. The performance of the multilayer films for sensing glucose could be tailored by controlling the gold nanoparticles loading in the film and the number of layers. A biosensor constructed by four poly(dimethyldiallylammonium chloride) (PDDA)/AuNP/PDDA/GOx multilayer films exhibited a wide linear calibration range from 0 to 60.0 mM with the detection limit of 3.0 microM for the detection of glucose. The dynamic range can be extended up to 120 mM. The biosensor has good stability and reproducibility.

Absorption↗