Biomedical subjects

Guoli Shen

Publications and source records attributed to Guoli Shen.

At least 19 recordsLinked to original sources

PubMed2006

Rapid, simple, and sensitive immunoagglutination assay with SiO2 particles and quartz crystal microbalance for quantifying Schistosoma japonicum antibodies.

BACKGROUND: The resurgence of the parasitic disease schistosomiasis calls for more efficient diagnostic tests. We developed a rapid, simple, portable, and sensitive immunoagglutination assay that uses SiO(2) particles and quartz crystal microbalance (QCM) for quantifying Schistosoma japonicum (Sj) antibodies (SjAb). METHODS: We prepared submicrometer-sized silica particles derivatized with Sj antigens as replacements for traditional latex microspheres to specifically agglutinate in the presence of SjAb targets, and we used the QCM monitor to measure the resulting frequency shifts. We optimized the assay medium by adding poly(ethylene glycol) (PEG) as a response accelerator of immunoagglutination. To minimize or eliminate any nonspecific agglutination or adsorption interferences, we conducted appropriate sealing procedures separately for silica particles and the QCM probe. RESULTS: The measured frequency changes were linearly related to the SjAb concentrations in infected rabbit serum. The PEG-assisted immunoagglutination system was quantitatively sensitive to SjAb concentrations ranging from approximately 0.70 to 32.31 mg/L, with a detection limit of approximately 0.46 mg/L. The obtained linear regression equation was: y=43.61 x+80.44 (r=0.9872). Several serum specimens were evaluated with the developed QCM immunoassay and the results were compared with ELISA, validating the feasibility of practical applications. CONCLUSIONS: This novel immunoagglutination-based QCM detection format is rapid, simple to use, and more portable than conventional diagnostic immunoassays, thus offering a promising alternative tool that can be used for point-of-care clinical diagnosis of schistosomiasis, particularly in epidemic situations.

Agglutination Tests↗

PubMed2006

Electrochemical biosensing utilizing synergic action of carbon nanotubes and platinum nanowires prepared by template synthesis.

Platinum nanowires (PtNWs) prepared by electrodeposition method with the help of porous anodic aluminum oxide (AAO) templates have been solubilized in chitosan (CHIT) together with carbon nantubes (CNTs) to form a PtNW-CNT-CHIT organic-inorganic system. The resulting PtNW-CNT-CHIT material brings capabilities for utilizing synergic action of PtNWs and CNTs to facilitate electron-transfer process in electrochemical sensor design. The PtNW-CNT-CHIT film modified electrode offered a significant decrease in the overvoltage for the hydrogen peroxide and showed to be excellent amperometric sensors for hydrogen peroxide at -0.1 V over a wide range of concentrations, and the sensitivity is 260 microAmM-1cm-2. As an application example, by linking glucose oxidase (GOx), an amplified biosensor toward glucose was prepared. The glucose biosensor exhibits a selective determination of glucose at -0.1 V with a linear response range of 5 x 10(-6) to 1.5 x 10(-2)M with a correlation coefficient of 0.997, and response time <10s. The high sensitivity of the glucose biosensor is up to 30 microAmM-1cm-2 and the detection limit was 3 microM. The biosensor displays rapid response and expanded linear response range, and excellent repeatability and stability.

Biosensing Techniques↗

PubMed2006

Platinum nanowire nanoelectrode array for the fabrication of biosensors.

Platinum nanowire arrays can be grown by electrodeposition in polycarbonate membrane, with the average diameter of the nanowires about 250 nm and the height about 2 microm. The nanowire array prepared by the proposed method can be considered as nanoelectrode array (NEA) with nanoelectrode density of 5 x 10(8)cm(-2). While the NEA can improve the signal-to-noise ratio and decrease the detection limit, the high surface area of the platinum NEA circumvents the problem of conventional platinum electrodes associated with the limited electroactive site. The platinum NEA can direct response to hydrogen peroxide at low potential of 0 V with wide linear range (1 x 10(-7)-6 x 10(-2)M) and sensitivity 50 times larger than that of the conventional platinum electrode. With the absorption of glucose oxidase onto the ordered NEA surface, the spatially patterned glucose oxidase improves greatly the resulting biosensor. The biosensor can achieve interference free determination of glucose with wide linear range (10(-6)-3 x 10(-2)M). The sensitivity of the glucose biosensor is one-fifth of the sensitivity toward hydrogen peroxide, indicating high efficiency of signal transduction. The biosensor was used to determine glucose in real blood samples, and the glucose contents determined by the present biosensor were in agreement with the results of existing method.

Biosensing Techniques↗

PubMed2006

Amperometric determination of bovine insulin based on synergic action of carbon nanotubes and cobalt hexacyanoferrate nanoparticles stabilized by EDTA.

A simple approach is proposed for the synthesis of cobalt hexacyanoferrate nanoparticles (CoNPs) with uniform shape and size controlled by ethylene diamine tetraacetic acid (EDTA) as a stabilizer. A sensitive amperometric biosensor for insulin has been prepared using glassy carbon electrodes by solubilization of carbon nanotubes (CNTs) in chitosan (CHIT) together with CoNPs synthesized by the new methodology. The CoNP-CNT-CHIT organic-inorganic system exerts a synergistic effect, resulting in the remarkably enhanced insulin currents owing to the superior electron-transfer ability of CNTs and the excellent reversible redox centers of CoNPs. High-resolution transmission electron microscopy (HRTEM) was used to provide closer inspection of the CoNPs. The effects of alkali metal cations and the concentrations of CNTs and CoNPs on the voltammetric behavior of the film-modified electrode were also investigated. In pH 6.98 phosphate buffer (PB) at +0.7 V (vs. SCE) the insulin biosensor exhibits a linear response range of 0.1-3 microM with a correlation coefficient of 0.98, and the detection limit (S/N=3) is determined to be 40 nM, the stability of the biosensor was tested and found satisfactory. There is great promise for in vivo measurements of this important hormone.

Amino Acid Sequence↗

PubMed2006

DNA point mutation detection based on DNA ligase reaction and nano-Au amplification: a piezoelectric approach.

A novel piezoelectric method for DNA point mutation detection based on DNA ligase reaction and nano-Au-amplified DNA probes is proposed. A capture probe was designed with the potential point mutation site located at the 3' end and a thiol group at the 5' end to be immobilized on the gold electrode surface of quartz crystal microbalance (QCM). Successive hybridization with the target DNA and detection probe of nano-Au-labeled DNA forms a double-strand DNA (dsDNA). After the DNA ligase reaction and denaturing at an elevated temperature, the QCM frequency would revert to the original value for the target with single-base mismatch, whereas a reduced frequency response would be obtained for the case of the perfect match target. In this way, the purpose of point mutation discrimination could be achieved. The current approach is demonstrated with the identification of a single-base mutation in artificial codon CD17 of the beta-thalassemia gene, and the wild type and mutant type were discriminated successfully. The scanning electron microscope (SEM) image showing that plenty of gold nanoparticles remained on the electrode surface demonstrated that the nano-Au label served as an efficient signal amplification agent in QCM assay. A detection limit of 2.6 x 10(-9)mol/L of oligonucleotides was achieved. Owing to its ease of operation and low detection limit, it is expected that the proposed procedure may hold great promise in both research-based and clinical genomic assays.

Base Pair Mismatch↗

PubMed2006

Immunophenotyping of acute leukemias using a quartz crystal microbalance and monoclonal antibody-coated magnetic microspheres.

Immunophenotyping, which utilizes panels of lineage-associated monoclonal antibodies to recognize clusters of differentiation (CD) antigens expressed on various leukocytes, plays a key role in the clinical diagnosis of acute leukemias. In this paper, a rapid, simple, and automatic immunophenotyping technique for acute leukemias has been initially proposed by incorporating immunomagnetic separation and quartz crystal microbalance (QCM) measurement. Core-shell paramagnetic microspheres of silica-coated ZnFe2O4 were newly fabricated following a W/O microemulsion route and further functionalized for anchoring separately CD antibodies of lineage-different leukocytes. The resultant immunomagnetic microspheres were introduced, in turn, to capture target leukocytes from the samples and were magnetically accumulated onto the protein A-modified QCM to be analyzed. The response characteristics of the developed QCM system for immunophenotyping various lineage-defined leukocytes were investigated in detail. Results indicate that this new technique can allow for easy and clear identification of acute leukocytes of lymphoid and myeloid origins as well as their subsets. It may also permit the quantitative determination of acute leukocytes with cell concentration down to approximately 10(3) cells mL(-1). Moreover, the applicable feasibility of the QCM immunophenotyping method was validated through assessing a number of clinical specimens, which phenotypes are in acceptable agreement with those obtained by the immunoenzyme assay clinically used for this purpose.

Acute Disease↗

PubMed2006

Orientation of 6-mercaptopurine SAMs at the silver electrode as studied by Raman mapping and in situ SERS.

Self-assembled monolayers (SAMs) of 6-mercaptopurine (6MP) on a silver electrode in acid and alkaline media were investigated by a combination protocol of the SERS technique with Raman mapping, and it was found that the adsorption mode of 6MP SAMs changed with the pH value of the environment. Quantum calculations for the vibrational mode were performed by the BLYP/6-31G method. 6MP was adsorbed on the silver electrode with a tilted orientation via S, N1, and N7 atoms in acid medium, while the SAMs adopted head-on adsorption modes with the S atom and the N1 atom anchoring the silver surface in alkaline medium. However, 6MP SAMs turned to the same upright orientation on the electrode through the S and N7 atoms when either acid or basic solution was removed. Stability of 6MP SAMs was observed by in situ SERS spectroelectrochemical measurements. The results reveal that the desorption potentials of 6MP SAMs formed under acid and alkaline conditions from the Ag electrode were at ca. -1.3 V and -1.6 V vs SCE, respectively.

Adsorption↗

PubMed2006

An amplified mass piezoelectric immunosensor for Schistosoma japonicum.

An ultrasensitive piezoelectric immunosensor using an amplification path based on an insoluble biocatalyzed precipitation product has proposed for Schistosoma japonicum. A mercapto Schistosoma japonicum antigen was self-assembled onto the quartz crystal surface via an Au nanoparticle mediator monolayer to sense the Schistosoma japonicum antibody (SjAb). And the horseradish peroxidase labeled protein A conjugate which was bounded to the SjAb by a "sandwich" format was used as a biocatalyst for the oxidative precipitation of 4-chloro-1-naphthol by H(2)O(2) to yield the insoluble product benzo-4-chlorohexadienone, resulting in an amplified mass sensing of antigen-antibody interaction. The amount of the precipitate accumulated on the quartz crystal is controlled by the antibody concentration. The SjAb can be linearly determined in the range of 10-200 ngml(-1) and the detection limit reaches as low as 5 ngml(-1).

Animals↗

PubMed2006

Platinum nanoparticles-doped sol-gel/carbon nanotubes composite electrochemical sensors and biosensors.

Platinum nanoparticle-doped sol-gel solution is prepared and used as a binder for multi-walled carbon nanotubes (CNT) for the fabrication of electrochemical sensors. Amine group containing sol-gel solution is selected to utilize the affinity of -NH(2) groups toward metal nanoparticles for stabilization the nanoparticles in solution. The resulting CNT-silicate material brings new capabilities for electrochemical devices by using the synergistic action of the electrocatalytic activity of Pt nanoparticles and CNT. The combined electrocatalytic activity permits low-potential detection of hydrogen peroxide with remarkably improved sensitivity. With the incorporation of glucose oxidase within the Pt-CNT-silicate matrix, a Pt-CNT paste-based biosensor has been constructed that responds more sensitively to glucose than CNT-based biosensor. The influences of the composite of the sol-gel solution, the quantity of the solution and the Pt nanoparticles loading are examined. In pH 6.98 phosphate buffer, almost interference free determination of glucose is realized at 0.1 V versus SCE with a linear range from 1 to 25 mM, a response time <15s, and the sensitivity is 0.98 microA mM(-1)cm(-2). The sensitivity of the Pt-CNT paste-based biosensor is almost four times larger than that of the CNT-based biosensor (0.27 microA mM(-1)cm(-2) at 0.1 V). The improved electrocatalytic activity and surface renewability made the Pt-CNT-silicate system a potential platform to immobilize different enzymes for other bioelectrochemical applications.

Adsorption↗

PubMed2006

Development of quartz-crystal-microbalance-based immunosensor array for clinical immunophenotyping of acute leukemias.

An integrated piezoelectric immunosensor array has been developed to immunophenotype acute leukemias in clinic. Each quartz crystal microbalance (QCM) was fabricated with plasma-polymerized film of n-butylamine, nanogold particles, and protein A (PA) to be used to immobilize antibodies in orientation. Leukemic lineage-associated monoclonal antibodies were separately immobilized onto the nanogold-PA-modified surface of the crystals, which were constructed by a 2 x 2 type of probes forming a QCM-based immunosensor array. The main detection conditions were investigated, including the immobilization amount of antibodies, pH, immunoreaction time, sample dilution ratio, etc. The immunophenotyping feasibility of the new technique was investigated through simultaneously analyzing Jurkat cells by the immunosensor array method, immunohistochemistry, and flow cytometry. It was found that the developed technique could readily identify leukemia samples in 5 min and might monitor dynamically the immunoreaction processes. Moreover, comparison studies were carried out for CD antigens expressed on the nucleated cells isolated from 96 acute leukemic patients and 24 normal subjects using the QCM-based immunosensor method and the fluoroimmunoassay. Results obtained by immunophenotyping patients' samples with the immunosensor-based method achieved the rate of 88.93% in 768 groups of numerical data, where no significant statistical difference was observed between the two methods when checked by chi2 analysis (chi2 = 3.4, p > 0.05). This new immunosensor array showed the merits of high sensitivity, high specificity, good reproducibility, easy operation, and low cost. The results of specimen evaluation indicated that it might be clinically suitable for quantifying human differentiated leukocytes and immunophenotyping of acute leukemias.

Acute Disease↗

PubMed2006

Layer-by-layer self-assembled multilayer films of carbon nanotubes and platinum nanoparticles with polyelectrolyte for the fabrication of biosensors.

Platinum nanoparticle-doped chitosan (CHIT) solution can be easily prepared by treating the CHIT solution with aqueous H2PtCl6 solution followed by chemical reduction of Pt(IV) with NaBH4. Multiwalled carbon nanotubes (MWCNT) are then dispersed in the nanoparticle-doped solution. The resulting Pt-CNT-CHIT material brings new capabilities for electrochemical devices by using the synergistic action of Pt nanaoparticles and CNT. Positively charged Pt-CNT-CHIT solution and negatively charged poly(sodium-p-styrenesulfonate) salt (PSS) have been employed to fabricate stable ultrathin multilayer films on gold electrode and quartz glass slides in a layer-by-layer fashion. Cyclic voltammetric and UV-vis adsorption spectroscopy confirms the consecutive growth of the multilayer films. The modified gold electrode allows low-potential detection of hydrogen peroxide with high sensitivity and fast response time. With the immobilization of cholesterol oxidase onto the electrode surface using glutaric dialdehyde, a biosensor that responds sensitively to cholesterol has been constructed. In pH 6.98 phosphate buffer, almost interference free determination of cholesterol has been realized at 0.1 V vs. SCE with a linear range from 0.01 to 3mM and response time<30s. With the immobilization of another cholesterol esterase enzyme layer, the biosensor was used to determine total cholesterol in serum samples with satisfactory results.

Biosensing Techniques↗

PubMed2005

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

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

Adsorption↗

PubMed2005

A colorimetric method for point mutation detection using high-fidelity DNA ligase.

The present study reported proof-of-principle for a genotyping assay approach that can detect single nucleotide polymorphisms (SNPs) through the gold nanoparticle assembly and the ligase reaction. By incorporating the high-fidelity DNA ligase (Tth DNA ligase) into the allele-specific ligation-based gold nanoparticle assembly, this assay provided a convenient yet powerful colorimetric detection that enabled a straightforward single-base discrimination without the need of precise temperature control. Additionally, the ligase reaction can be performed at a relatively high temperature, which offers the benefit for mitigating the non-specific assembly of gold nanoparticles induced by interfering DNA strands. The assay could be implemented via three steps: a hybridization reaction that allowed two gold nanoparticle-tagged probes to hybrid with the target DNA strand, a ligase reaction that generates the ligation between perfectly matched probes while no ligation occurred between mismatched ones and a thermal treatment at a relatively high temperature that discriminate the ligation of probes. When the reaction mixture was heated to denature the formed duplex, the purple color of the perfect-match solution would not revert to red, while the mismatch gave a red color as the assembled gold nanoparticles disparted. The present approach has been demonstrated with the identification of a single-base mutation in codon 12 of a K-ras oncogene that is of significant value for colorectal cancers diagnosis, and the wild-type and mutant type were successfully scored. To our knowledge, this was the first report concerning SNP detection based on the ligase reaction and the gold nanoparticle assembly. Owing to its ease of operation and high specificity, it was expected that the proposed procedure might hold great promise in practical clinical diagnosis of gene-mutant diseases.

Cell Line↗

PubMed2005

Carbon nanotube/cobalt hexacyanoferrate nanoparticle-biopolymer system for the fabrication of biosensors.

Cobalt hexacyanoferrate nanoparticles (CoNP) can be easily prepared by mixing hexacyanoferrate and cobalt chloride solution at room temperature. The nanoparticles were solubilized in aqueous solution of a biopolymer chitosan (CHIT). With the introduction of carbon nanotubes (CNT), the CoNP-CNT-CHIT system formed shows synergy between CNT and CoNP with the significant improvement of redox activity of CoNP due to the excellent electron-transfer ability of CNT. The CoNP-CNT-CHIT film modified glassy carbon electrode allows low potential detection of hydrogen peroxide with high sensitivity and fast response time. In particular, with the introduction of CNT, it amplified the H2O2 sensitivity by approximately 70 times compared to film of CoNP-CHIT. With the immobilization of glucose oxidase onto the electrode surface using glutaric dialdehyde, a biosensor that responds sensitively to glucose has been constructed. In pH 6.98 phosphate buffer, interference free determination of glucose has been realized at -0.2V versus saturated calomel electrode (SCE) with a linear range from 0.01 to 10 mM and response time<10s. The detection limit was 5 microM glucose (S/N=3).

Biocompatible Materials↗

PubMed2005

Amperometric biosensor for choline based on layer-by-layer assembled functionalized carbon nanotube and polyaniline multilayer film.

Conducting polymer film was prepared by electrochemical polymerization of aniline. Multiwalled carbon nanotubes (MWNTs) were treated with a mixture of concentrated sulfuric and nitric acid to introduce carboxylic acid groups to the nanotubes. By using the layer-by-layer method, homogeneous and stable MWNTs and polyaniline (PANI) multilayer films were alternately assembled on glassy carbon (GC) electrodes. Conducting polymer of PANI had three main functions: (i) excellent antiinterference ability, (ii) protection ability in favor of increasing the amount of the MWNTs immobilized on GC electrodes, and (iii) superior transducing ability. The protection effect of PANI film and the electrostatic interaction between positively charged PANI and negatively charged MWNTs both attributed to immobilizing abundant MWNTs stably, thereby enhancing the catalytic activity. The layer-by-layer assembled MWNTs and PANI-modified GC electrodes offered a significant decrease in the overvoltage for the H2O2 and were shown to be excellent amperometric sensors for H2O2 from +0.2V over a wide range of concentrations. As an application example, by linking choline oxidase (CHOD), an amplified biosensor toward choline was prepared. The choline biosensor exhibited a linear response range of 1x10(-6) to 2x10(-3) M with a correlation coefficient of 0.997, and the response time and detection limit (S/N=3) were determined to be 3 s and 0.3 microM, respectively. The antiinterference biosensor displays a rapid response and an expanded linear response range as well as excellent reproducibility and stability.

Alcohol Oxidoreductases↗

PubMed2005

Enzyme-catalyzed amplified immunoassay for the detection of Toxoplasma gondii-specific IgG using Faradaic impedance spectroscopy, CV and QCM.

A highly sensitive electrochemical immunoassay for Toxoplasma gondii-specific IgG (Tg-IgG) in human serum has been developed that is based on an enzyme-catalyzed amplification due to the formation of an insoluble precipitate on the surface of a quartz crystal microbalance (QCM). T. gondii antigen (TgAg) was immobilized on the surface of a gold electrode in order to bind Tg-IgG, and this was followed by the addition of anti-Tg-IgG horseradish peroxidase conjugate (anti-Tg-IgG-HRP). Subsequent exposure to 3,3'-diaminobenzidine (DAB) led to the enzymatically-catalyzed amplified deposition of the oxidation products on the QCM surface in the presence of H2O2. The transduction methods electrochemical Faradaic impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used to assay the resistance to electron transfer at the conductive support upon accumulation of the insoluble products. The precipitation process was monitored in real time by QCM. The assay conditions, including the concentration of immobilized TgAg and the dosage of anti-Tg-IgG-HRP conjugate, were optimized. It was found that the amount of precipitate that accumulated on the conductive QCM surface was determined by the concentration of the target analyte Tg-IgG and the time permitted for biocatalyzed precipitation. The technique was shown to give a linear electron transfer resistance response (as measured by EIS) for Tg-IgG dilutions ranging between 1:8000 and 1:200, and a detection limit of 1:9600 dilution.

Animals↗

PubMed2005

Nanogold particle-enhanced oriented adsorption of antibody fragments for immunosensing platforms.

A general design strategy for immunosensing platforms has been proposed on the basis of Nanogold particle-enhanced oriented adsorption of antibody fragments. Quartz crystal microbalance (QCM) as a model transducer was fabricated with plasma-polymerized film (PPF) of n-butyl amine and then with nanogold particles resulting in a PPF-nanogold adsorption procedure for half-IgG fragments obtained by reduction of intact immunoglobulin (IgG). Thermodynamic studies reveal that the proposed procedure is superior to the traditional oriented ones in that it created immunosurface of increased antibody surface density (amount) and antigen binding constants. Sensors produced according to the new immobilization procedure exhibit better immunosensing performances including high sensitivity, fast response rate, and favorable operational stability etc. This Nanogold particle-enhanced immobilization technique may be tailored as a promising alternative for various immunosensing platforms in solid-phase immunoassay and affinity chromatography.

Adsorption↗

PubMed2005

Raman mapping and in situ SERS spectroelectrochemical studies of 6-mercaptopurine SAMs on the gold electrode.

The self-assembled monolayers (SAMs) of 6-mercaptopurine (6MP) were formed at the roughened polycrystalline gold surfaces in acid and alkaline media. The time-dependent Raman mapping spectral analysis in conjunction with the quantum calculations for the vibrational modes using ab initio BLYP/6-31G method suggested that both of the resulted 6MP SAMs adopted the same adsorption mode through the S atom of pyrimidine moiety and the N7 atom of the imidazole moiety anchoring the gold surface in a vertical way. The in situ surface-enhanced Raman scattering spectroelectrochemical experiment was conducted to examine the stability of the SAMs at various bias potentials. It was found that the detaching process of the 6MP SAMs from the surface involved one electron reduction as the voltage was applied at ca. 0.7 V vs a standard calomel electrode.

Biophysics↗