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1.
《Electroanalysis》2018,30(3):402-414
A sensitive electrochemical immunosensor for Hepatitis B virus surface antigen (HBsAg) detection was fabricated based on hemin/G‐quadruplex interlaced onto Fe3O4‐AuNPs or hemin ‐amino‐reduced graphene oxide nanocomposite (H‐amino‐rGO‐Au). G‐quadruplex DNAzyme, which is composed of hemin and guanine‐rich nucleic acid, is an effective signal amplified tool for its outstanding peroxidase activity and Fe3O4‐AuNPs or (H‐amino‐rGO‐Au) nanocomposites with quasi‐enzyme activity provide appropriate support for the immobilization of hemin/G‐quadruplex. The target protein was sandwiched between the primary antibody immobilized on the GO and secondary antibody immobilized on the Fe3O4‐AuNPs or (H‐amino‐rGO‐Au) nanocomposites and glutaraldehyde was used as linking agent for the immobilization of primary antibody on the surface of GO. Both Fe3O4‐AuNPs and H‐amino‐rGO‐Au nanocomposite and also hemin/G‐quadruplex can cooperate the electrocatalytic reduction of H2O2 in the presence of methylene blue as mediator. The proposed immunosensor has a wide linear dynamic range of 0.1 pg/ml to 300 pg/ml with a detection limit of 60 fg/ml when Fe3O4‐AuNPs was used for immobilization of hemin/G‐quadruplex, while the dynamic range and DL were 0. 1–1000 pg/mL and 10 fg/mL, respectively in the presence of H‐amino‐rGO‐ Au nanocomposite as platform for immobilizing of hemin/G‐quadruplex. The proposed immunosensor was also used for analysis of HBsAg in spiked human serum samples with satisfactory results.  相似文献   

2.
The efficient electron injection by direct dye‐to‐TiO2 charge transfer and strong adhesion of mussel‐inspired synthetic polydopamine (PDA) dyes with TiO2 electrode is demonstrated. Spontaneous self‐polymerization of dopamine using dip‐coating (DC) and cyclic voltammetry (CV) in basic buffer solution were applied to TiO2 layers under a nitrogen atmosphere, which offers a facile and reliable synthetic pathway to make the PDA dyes, PDA‐DC and PDA‐CV, with conformal surface and perform an efficient dye‐to‐TiO2 charge transfer. Both synthetic methods led to excellent photovoltaic results and the PDA‐DC dye exhibited larger current density and efficiency values than those in the PDA‐CV dye. Under simulated AM 1.5 G solar light (100 mW cm?2), a PDA‐DC dye exhibited a short circuit current density of 5.50 mW cm?2, corresponding to an overall power conversion efficiency of 1.2 %, which is almost 10 times that of the dopamine dye‐sensitized solar cell. The PDA dyes showed strong adhesion with the nanocrystalline TiO2 electrodes and the interface engineering of a dye‐adsorbed TiO2 surface through the control of the coating methods, reaction times and solution concentration maximized the overall conversion efficiency, resulting in a remarkably high efficiency.  相似文献   

3.
《Electroanalysis》2018,30(5):834-841
We have investigated different surface functionalization methods to immobilize CD19 antibody on gold surface to capture B lymphoblast cells associated with the acute lymphoblastic leukemia disease. Quartz Crystal Microbalance measurements were performed to analyze the binding kinetics of each layer and determine the optimum method, which results in higher cell capture rates. The random orientation of antibody and oriented antibody through protein G was investigated and protein G presence resulted in 15,2 Hz frequency shift for 104 cells/mL. The 3‐mercaptopropyltrimethoxysilane (MPS) and 11‐Mercaptoundecanoic acid (MUA) coatings of gold surface together with 4‐(N‐Maleimidomethyl)cyclohexane‐1‐carboxylic acid 3‐sulfo‐N‐hydroxysuccinimide ester sodium salt (Sulfo‐SMCC) and N‐Ethyl‐N’‐(3‐dimethylaminopropyl) carbodiimide hydrochloride (EDC)/N‐hydroxysulfosuccinimide (NHS) linker layers were tested on QCM for protein G and antibody binding. The results indicate that MUA, EDC/NHS, protein G, antibody CD19 is the optimum surface modification among the tested combinations. By using the optimum surface functionalization method, minimum 103 cell per mL was measured as 1.9 Hz frequency shift.  相似文献   

4.
《化学:亚洲杂志》2017,12(16):2033-2037
The immunochromatographic assay (ICA) using a nitrocellulose (NC) membrane offers several advantages. This technique is a rapid and straightforward method in contrast to other immunoassays. Polydiacetylene (PDA) vesicles have unique optical properties, displaying red color and red fluorescence at the same time. In this system, red‐phase PDA vesicles are used as a fluorescent dye as well as a surface for immobilized hepatitis B surface antibody (HBsAb). PDA has a remarkable stability compared with other fluorescent dyes. In this study, the most suitable PDA/HBsAb complexes are introduced for detecting hepatitis B surface antigen (HBsAg). Then, the PDA/HBsAb complexes affixed antibody is attached to NC membrane, which has two lines to confirm detection of HBsAg. The main advantage of this system is that the detection of HBsAg can be observed in both visible and fluorescent images due to the optical properties of polydiacetylene. Detection of HBsAg is observed up to 0.1 ng mL−1 by fluorescent analysis and confirmed by red line on the NC membrane up to 1 ng mL−1 (HBsAg) using the naked eye. Consequently, these results show that PDA/HBsAb complexes were successfully applied to ICA for the diagnosis of hepatitis B.  相似文献   

5.
We have applied a reusable silicon nanowire field‐effect transistor (SiNW‐FET) as a biosensor to conduct ultrasensitive detection of H5N2 avian influenza virus (AIV) in very dilute solution. The reversible surface functionalization of SiNW‐FET was made possible using a disulfide linker. In the surface functionalization, 3‐mercaptopropyltrimethoxysilane (MPTMS) was first modified on the SiNW‐FET (referred to as MPTMS/SiNW‐FET), with subsequent dithiothreitol washing to reduce any possible disulfide bonding between the thiol groups of MPTMS. Subsequently, receptor molecules could be immobilized on the MPTMS/SiNW‐FET by the formation of a disulfide bond. The success of the reversible surface functionalization was verified with fluorescence examination and electrical measurements. A surface topograph of the SiNW‐FET biosensor modified with a monoclonal antibody against H5N2 virus (referred to as mAbH5/SiNW‐FET) after detecting approximately 10?17 M H5N2 AIVs was scanned by atomic force microscopy to demonstrate that the SiNW‐FET is capable of detecting very few H5N2 AIV particles.  相似文献   

6.
Molecular modeling, electrochemical methods, and quartz crystal microbalance were used to characterize immobilized hexameric tyrosine‐coordinated heme protein (HTHP) on bare carbon or on gold electrodes modified with positively and negatively charged self‐assembled monolayers (SAMs), respectively. HTHP binds to the positively charged surface but no direct electron transfer (DET) is found due to the long distance of the active sites from the electrode surfaces. At carboxyl‐terminated surfaces, the neutrally charged bottom of HTHP can bind to the SAM. For this “disc” orientation all six hemes are close to the electrode and their direct electron transfer should be efficient. HTHP on all negatively charged SAMs showed a quasi‐reversible redox behavior with rate constant ks values between 0.93 and 2.86 s?1 and apparent formal potentials ${E{{0{^{\prime }}\hfill \atop {\rm app}\hfill}}}$ between ‐131.1 and ‐249.1 mV. On the MUA/MU‐modified electrode, the maximum surface concentration corresponds to a complete monolayer of the hexameric HTHP in the disc orientation. HTHP electrostatically immobilized on negatively charged SAMs shows electrocatalysis of peroxide reduction and enzymatic oxidation of NADH.  相似文献   

7.
The Wilkinson’s catalyst [RhCl(PPh3)3] has been immobilized inside the pores of amine functionalized mesoporous silica material SBA‐3 and The structure of the modified silica surface and the immobilized rhodium complex was determined by a combination of different solid‐state NMR methods. The successful modification of the silica surface was confirmed by 29Si CP‐MAS NMR experiments. The presence of the Tn peaks confirms the successful functionalization of the support and shows the way of binding the organic groups to the surface of the mesopores. 31P‐31P J‐resolved 2D MAS NMR experiments were conducted in order to characterize the binding of the immobilized catalyst to the amine groups of the linkers attached to the silica surface. The pure catalyst exhibits a considerable 31P‐31P J‐coupling, well resolvable in 2D MAS NMR experiments. This J‐coupling was utilized to determine the binding mode of the catalyst to the linkers on the silica surface and the number of triphenylphosphine ligands that are replaced by coordination bonds to the amine groups. From the absence of any resolvable 31P‐31P J‐coupling in off‐magic‐angle‐spinning experiments, as well as slow‐spinning MAS experiments, it is concluded, that two triphenylphosphine ligands are replaced and that the catalyst is bonded to the silica surface through two linker molecules.  相似文献   

8.
In our photo‐induced immobilization technique for an antibody (IgG) using azopolymers, the introduction of COOH and NMe2 into the azopolymers, which can introduce surface charges, strongly affected the immobilization properties such as the efficiency of immobilization and the activity of the immobilized IgG (i.e., the orientation of the immobilized IgG). The introduction of COOH promoted a more active orientation of the immobilized IgG. The orientation was determined during the adsorption process onto the azopolymer surface in solution before photo‐immobilization, and was maintained during the photo‐immobilization. The surface charge of the azopolymer appears to be an important factor for IgG orientation, which involves electrostatic interactions between its Fab and the azopolymer surface.

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9.
Continuous outbreaks of avian influenza (AI) in recent years with increasing threat to animals and human health have warranted the urgent need for rapid detection of pathogenic AI viruses. In this study, an impedance immunosensor based on an interdigitated array (IDA) microelectrode was developed as a new application for sensitive, specific and rapid detection of avian influenza virus H5N1. Polyclonal antibodies against AI virus H5N1 surface antigen HA (Hemagglutinin) were oriented on the gold microelectrode surface through protein A. Target H5N1 viruses were then captured by the immobilized antibody, resulting in a change in the impedance of the IDA microelectrode surface. Red blood cells (RBCs) were used as biolabels for further amplification of the binding reaction of the antibody-antigen (virus). The binding of target AI H5N1 onto the antibody-modified IDA microelectrode surface was further confirmed by atomic force microscopy. The impedance immunosensor could detect the target AI H5N1 virus at a titer higher than 103 EID50/ml (EID50: 50% Egg Infective Dose) within 2 h. The response of the antibody-antigen (virus) interaction was shown to be virus titer-dependent, and a linear range for the titer of H5N1 virus was found between 103 and 107 EID50/ml. Equivalent circuit analysis indicated that the electron transfer resistance of the redox probe [Fe(CN)6]3−/4− and the double layer capacitance were responsible for the impedance change due to the protein A modification, antibody immobilization, BSA (bovine serum albumin) blocking, H5N1 viruses binding and RBCs amplification. No significant interference was observed from non-target RNA viruses such as Newcastle disease virus and Infectious Bronchitis disease virus. (The H5N1 used in the study was inactivated virus.)  相似文献   

10.
The complex [N‐(2,6‐diisopropylphenyl)‐2‐(2,6‐diisopropylphenylimino)‐propanamidato‐κ2N,O](η1‐benzyl)nickel(lutidine) is immobilized on an acid‐treated montmorillonite. When the montmorillonite is treated with alkylaluminum, the catalyst shows a high activity with high molecular‐weight polyethylene. The catalytic behavior of the polymerization suggests isomerization of the complex on acid sites of the support surface.  相似文献   

11.
Herein the stereoselective two‐step synthesis of pure exo‐5‐trimethylsilylnorbornene is reported. The monomer proved to be highly reactive in both metathesis and addition polymerization. ROMP polymerization was catalyzed by the first‐generation Grubbs catalyst. High‐molecular‐weight saturated addition polymers were prepared using nickel or palladium complexes as precatalysts and Na+[B(3,5‐(CF3)2C6H3)4] and/or MAO as cocatalysts. The obtained addition polynorbornenes are highly gas permeable and microporous materials possessing large free volume and BET surface area (up to 540 m2/g). The influence of the substituent orientation (exo‐ vs. exo‐/endo‐mixture) on polymer properties was established. The metathesis polymer based on exo‐isomer exhibits 1.5‐ to 2‐fold increase of permeability coefficients for all gases in comparison to the similar polymer based on the mixture of exo‐ and endo‐isomers. © 2018 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2018 , 56, 1234–1248  相似文献   

12.
A novel and sensitive electrochemiluminescence (ECL) method for ethanol biosensor was developed by co‐immobilizing the enzyme and ECL reagent Ru(bpy)32+ on the poly‐(3,4‐ethylene dioxythiophene) and polystyrene sulfonate functionalized graphene (PEDOT‐PSS‐G) nanocomposite film. Positively charged Ru(bpy)32+ could be immobilized effectively on the electrode surface with the negatively charged PSS and graphene, which provided a stable ECL platform for further modification with the enzyme. Moreover, the introduction of PEDOT and graphene can be acted as a conducting pathway to accelerate the electron transfer due to the high conductivity. Such biosensor combined enzymatic selectivity with the amplification of PEDOT‐PSS‐G performed well with a wide linear range, high sensitivity and good stability. The sensing platform was successfully applied to determine the amounts of alcohol in real samples.  相似文献   

13.
本文研制了一种用金胶壳聚糖仿生膜来同时固定四甲基联苯胺(TMB)和酶标抗体的新型电化学免疫传感器,用于检测血清肿瘤标志物前列腺特异性抗原(PSA)的含量。固定的TMB作为电子传递媒介体,在扫速小于45 mV/s时,电极表现为一个表面控制过程,而在扫速大于45 mV/s时则表现为一个扩散控制过程。将固定有酶标抗体和TMB的免疫传感器与待测PSA抗原一起培育,在该传感器上形成的免疫复合物通过TMB-H2O2-HRP电化学体系进行了测定。在优化实验条件下,PSA的线性检测范围为5-30 ng·mL-1,检测限为1.0 ng·mL-1。该PSA免疫传感器制备方法简单,成本低廉,具有较好的稳定性和重现性。  相似文献   

14.
Magnesium has been suggested as a potential biodegradable metal for the usage as orthopaedic implants. However, high degradation rate in physiological environment remains the biggest challenge, impeding wide clinical application of magnesium‐based biomaterials. In order to reduce its degradation rate and improve the biocompatibility, micro‐arc oxidation coating doped with HA particles (MAO‐HA) was applied as the inner coating, and polydopamine (PDA) film was synthesized by dopamine self‐polymerization as the outer coating. The microstructure evolution of the coating was characterized using scanning electron microscopy (SEM), atomic force microscope (AFM), X‐ray diffraction analyses (XRD), Fourier transform infrared spectroscopy (FT‐IR), and X‐ray photoelectron spectroscopy (XPS). The results showed that PDA film had covered the entire surface of MAO‐HA coating and the pore size of MAO‐HA coating decreased. The root mean square (RMS) roughness of PDA/MAO‐HA coatings was approximately 106.46 nm, which was closer to the optimum surface roughness for cellular attachment as compared with MAO‐HA coatings. Contact angle measurement indicated that the surface wettability had been transformed from hydrophobic to hydrophilic due to the introduction of PDA. The PDA/MAO‐HA coatings exhibited better corrosion resistance in vitro, with the self‐corrosion potential increasing by 150 mV and the corrosion current density decreasing from 2.09 × 10?5 A/cm 2 to 1.46 × 10?6 A/cm 2 . In hydrogen evolution tests, the corrosion rates of the samples coated with PDA/MAO‐HA and MAO‐HA were 4.40 and 5.95 mm/y, respectively. MTS assay test and cell‐surface interactions experiment demonstrated that PDA/MAO‐HA coatings exhibited good cellular compatibility and could promote the adhesion and proliferation of MC3T3‐E1 cells.  相似文献   

15.
The crystal structure of the title compound, [Pt(C6H7N)2(C20H16N4)](PF6)2·C3H6O·0.5H2O, is composed of a bivalent square‐planar platinum(II) complex, two PF6 counter‐ions and solvent mol­ecules. The di‐2‐pyridylquinoxaline ligands are known to confer an `L shape' on square‐planar platinum(II) complexes, which also display inter­calating properties. The structural characterization reported here is a contribution to a wide‐ranging study focused on structural and dynamical analyses of these substrates, which may provide better insight into their biological mechanisms and activities. The expected `L‐shaped' skeleton of the metallic complex combined with the antiparallel orientation of substituted pyridines (anti conformation) generates chiral objects, found in the solid state as a racemic mixture.  相似文献   

16.
A renewable potentiometric immunosensor for detection of immunoglobulin G (IgG) has been developed by magnetic force attraction of Fe3O4 nanoparticles immobilized goat‐anti‐human IgG antibody. For preparing sensitive film of the sensor, cysteine was bonded on the nano‐Fe3O4 particles surface. The cysteine functionalized magnetic nanoparticles was attracted on a solid paraffin carbon paste electrode surface to covalently immobilize of anti‐immunoglobulin G (anti‐IgG) by employing a conventional glutaraldehyde‐crosslinking method. The immunosensor showed a specific response to human immunoglobulin G in the range of 0.1–1.2 ng/mL with a detection limit of 0.023 ng/mL. The immunosensor based on the magnetic nanoparticles was made easily by this method. It can be used expediently, renewed easily and low‐cost relatively. The renewable potentiometric immunosensor with better stability and higher sensitivity can be employed extensively in clinical diagnosis, monitoring of disease and environmental studies and etc.  相似文献   

17.
In the present study, we developed a novel label‐free capacitance impedimetric immunosensor based on the immobilization of the human monoclonal antibody anti‐interleukin‐10 (anti‐IL‐10 mAb) onto polypyrrole (PPy)‐modified silicon nitride (Si3N4) substrates. The immunosensor was used for the detection of the recombinant interleukin‐10 antigen (rh IL‐10) that may be secreted in patients at the early stage of inflammation. The immunosensor was created by chemical deposition of PPy conducting layer on pyrrole?silane (SPy)‐treated Si/SiO2/Si3N4 substrates (Si/SiO2/Si3N4?SPy), followed by anti‐IL‐10 mAb immobilization through carboxyl‐functionalized diazonium (CMA) protocol and carbodiimide chemistry. The surface characterization and the biofunctionalization steps were characterized by SEM, FTIR and cyclic voltammetry (CV) while the detection process was carried out by using electrochemical impedance spectroscopy (EIS) analyses. The created immunosensor showed two linear fittings (R2=0.999) for the detection of rh IL‐10 within the concentration range from 1–50 pg/mL. It exhibited high sensitivity (0.1128 (pg/mL)?1) with a very low limit of detection (LOD)=0.347 pg/mL, more particularly, at the low concentration range (1–10 pg/mL). Thus, this developed polypyrrole‐based immunosensor represents a promising strategy for creation of miniaturized label‐free, fast and highly sensitive biosensors for diagnosis of inflammation biomarkers at very low concentrations with reduced cost.  相似文献   

18.
Solvothermal reaction between Cd(NO3)2, 1,4‐phenylenediacetate (1,4‐PDA) and 1,3‐bis(pyridin‐4‐yl)propane (bpp) afforded the title complex, [Cd(C10H8O4)(C13H14N2)]n. Adjacent carboxylate‐bridged CdII ions are related by an inversion centre. The 1,4‐PDA ligands adopt a cis conformation and connect the CdII ions to form a one‐dimensional chain extending along the c axis. These chains are in turn linked into a two‐dimensional network through bpp bridges. The bpp ligands adopt an antigauche conformation. From a topological point of view, each bpp ligand and each pair of 1,4‐PDA ligands can be considered as linkers, while the dinuclear CdII unit can be regarded as a 6‐connecting node. Thus, the structure can be simplified to a two‐dimensional 6‐connected network.  相似文献   

19.
A dual‐genic impedimetric sensor for DNA hybridization detection is proposed for the first time. Two different single‐stranded DNA probes (P1=d(pA)20 and P2=d(pG)20) were immobilized onto the graphite‐epoxy electrode surface and impedimetric measurements were performed. Hybridization experiments were carried out employing complementary targets (P1c=d(pT)20 or P2c=d(pC)20) alone, or combined (P1c+P2c), as well as noncomplementary DNA sequences. As from the direct observation of impedance spectra it was not possible to resolve the binary gene mixture, the use of artificial neural networks (ANN) was proposed for extracting significant information to get desired results.  相似文献   

20.
N‐Formyl‐1‐bromo‐4‐hydroxy‐3‐methoxymorphinan‐6‐one (compound 2 ), an important intermediate in the NIH Opiate Total Synthesis, presumably exists as a mixture of two rotamers (Z and E) in both CHCl3 and DMSO at room temperature due to the hindered rotation of its N‐C18 bond in the amide moiety. By comparing the experimental 1H and 13C chemical shifts of a single rotamer and the mixture of compound 2 in CDCl3 with the calculated chemical shifts of the geometry optimized Z and E rotamers utilizing density functional theory, the crystalline rotamer of compound 2 was characterized as having the E configuration. The energy barrier between the two rotamers was also determined with the temperature dependence of 1H and 13C NMR coalescence experiments, and then compared with that from the reaction path for the interconversion of the two rotamers calculated at the level of B3LYP/6‐31G*. Detailed geometry of the ground state and the transition states of both rotamers are given and discussed. Copyright © 2012 This article is a US Government work and is in the public domain in the USA.  相似文献   

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