100 resultados para Indirect antibody competition ELISA


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The study of rotation-alignment of quasiparticles probes sensitively the properties of high-j intruder orbits. The distribution of very-high-j orbits, which are consequences of the fundamental spin-orbit interaction, links with the important question of single-particle levels in superheavy nuclei. With the deformed single-particle states generated by the standard Nilsson potential, we perform Projected Shell Model calculations for transfermium nuclei where detailed spectroscopy experiments are currently possible. Specifically, we study the systematical behavior of rotation-alignment and associated band-crossing phenomenon in Cf, Fm, and No isotopes. Neutrons and protons from the high-j orbits are found to compete strongly in rotation-alignment, which gives rise to testable effects. Observation of these effects will provide direct information on the single-particle states in the heaviest nuclear mass region.

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Silica-based functionalized terbium fluorescent nanoparticles were prepared, characterized and developed as a fluorescence probe for antibody labeling and time-resolved fluoroimmunoassay. The nanoparticles were prepared in a water-in-oil (W/O) microemulsion containing a strongly fluorescent Tb3+ chelate. N,N.N-1,N-1-12,6-bis(3'-aminomethyl-1'-pyrazolyl)phenylpyridine] tetrakis(acetate)-Tb3+ (BPTA-Tb3+), Triton X-100, octanol, and cyclohexane by controlling copolymerization of tetraethyl orthosilicate (TEOS) and 3-[2-(2- aminoethylamino)-ethylamino]propyl-trimethoxysilane (AEPS) with ammonia water. The characterizations by transmission electron microscopy and fluorometric quantum methods show that the nanoparticles are spherical and uniform in size, 45 +/- 3 nm in diameter, strongly fluorescent with fluorescence yield of 10% and a long fluorescence lifetime of 2.0 ms. The amino groups directly introduced to the nanoparticle's surface by using AEPS in the preparation made the surface modification and bioconjugation of the nanoparticles easier. The nanoparticle-labeled anti-human alpha-fetoprotein antibody was prepared and used for time-resolved fluoroimmunoassay of (x-fetoprotein (AFP) in human serum samples. The assay response is linear from 0.10 ng ml(-1) to about 100 ng ml(-1) with the detection limit of 0.10 ng ml(-1). The coefficient variations (CVs) of the method are less than 9.0%. and the recoveries are in the range of 84-98% for human serum sample measurements. (C) 2004 Elsevier B.V. All rights reserved.

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The development of a method for determining arsenic species by capillary zone electrophoresis (CZE) with indirect laser-induced fluorescence (LIF) is described in this paper. The buffer pH, the concentration of fluorescein, the nature and the concentration of the background electrolytes (BGEs) were defined. When 2.0 mM NaHCO3 (pH 9.28) with 10(-7) M fluorescein was used as the buffer, arsenite (As(lll), dimethylarsonic acid (DMA), monomethylarsonic acid (MMA), and arsenate (As(V)) were all separated from one another. The limits of detection for the four arsenic species were p p in the range of 0.12-0.54 mg/L. This method was used in the analysis of spiked arsenic species in tap and mineral water to demonstrate its usefulness. The results showed that both the recovery and the reproducibility of the developed method were acceptable.

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Langmuir-Blodgett (LB) film of stearic acid was used as template to induce the nucleation and growth of KCl crystals when the KCl solution was cooled from 50 to 25 degrees C. When the LB film template was vertically dipped into the solution, only induced crystals with (1 1 0) orientation were formed. However, if the template was horizontally placed into solutions, both the induced nuclei at the solution/film interface and spontaneous nuclei formed in solution were simultaneously absorbed onto the LB film, and then grew further to form crystals. X-ray diffraction (XRD) patterns and optical microscopy images showed that the orientation and morphology of the crystals were controlled properly by changing the orientation and position of the LB films in the solutions.

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We investigate the dewetting behavior of the bilayer of air/PS/PMMA/silanized Si wafer and find the two competing dewetting pathways in the dewetting process. The upper layer dewets on the lower layer (dewetting pathway 1, the liquid-liquid dewetting) and the two layers rupture on the solid substrate (dewetting pathway 2, the liquid-solid dewetting). To the two competing dewetting pathways, the process of forming holes and the process of hole growth, influence their competing relation. In the process of forming holes, the time of forming holes is a main factor that influences their competing relation. During the process of hole growth, the dewetting velocity is a main factor that influences their competing relation.

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A novel tris(2,2'-bipyridine)ruthenium(II) (Ru(bpy)3(2+)) cathodic electrochemiluminescence (ECL) was generated at -0.78V at the Pt electrode in acetonitrile (ACN), which suggested that the cathodic ECL differed from conventional cathodic ECL It was found that tripropylamine (TPrA) could enhance this cathodic ECL and the linear range (log-log plot) was 0.2 mu M-0.2 mM. In addition, hydrogen peroxide (H2O2) could inhibit the cathodic ECL and was indirectly detected with the linear range of 27-540 mu M. The RSD (n = 12) of the ECL intensity in the presence of 135 mu M H2O2 was 0.87%. This method was also demonstrated for the fast determination of H2O2 in disinfectant sample and satisfactory results were obtained.

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We have studied the lamellar orientation in thin films of a model diblock copolymer, symmetric poly(styrene)-b-poly(L-lactide) (PS-PLLA), in the melt state on supported silicon wafer surface. In this system, while the PLLA block prefers to wet the polymer/substrate interface, the polymer/air as well as polymer/polymer interface is neutral for both blocks due to the similar surface energies of PS and PLLA in melt state. Our results demonstrate that the interplay of the interfaces during phase separation results in a series of structures before approaching the equilibrium state. Lamellar orientation of thin films with different initial film thicknesses at different annealing stages has been investigated using atomic force microscopy (AFM), transmission electronic microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). It is found that in the early stage (annealing time t < 10 min), the polymer/substrate interface dominates the structure evolution, leading to a parallel lamellar structure with holes or islands formed depending on the initial film thickness. Later on, the neutral air interface becomes important and leads to a transition of lamellar orientation from parallel to perpendicular. It is interesting to see that for films with thickness h > 2L, where L is the bulk lamellar period, the lamellar orientation transition can occur independently in different parallel lamellar domains due to the neutrality of polymer/polymer interface.

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We used colloidal An to enhance the amount of antibody immobilized on a gold electrode and ultimately monitored the interaction of antigen-antibody by impedance measurement. Self-assembly of 6 nm (diameter) colloidal An onto the self-assembled monolayers (SAMs) of 4-aminothiophenol modified gold electrode resulted in an easier attachment of antibody. The redox reactions of [Fe(CN)(6)](4-)/[Fe(CN)(6)](3-) on the gold surface were blocked due to the procedures of self-assembly of 4-aminothiophenol and antibody immobilization, which were investigated by cyclic voltammetry and impedance spectroscopy. The interaction of antigen with grafted antibody recognition layers was carried out by soaking the modified electrode into a phosphate buffer at pH 7.4 with various concentrations of antigen at 37 degreesC for 30 min. The antibody recognition layers and their interactions with various concentrations of antigen could be detected by measurements of the impedance change. The results show that this method has good correlation for detection of Hepatitis B virus surface antigen in the range of 0.5-200 mug/l and a detection limit of about 50 ng/l.

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Gold nanoparticles were used to enhance the immobilization amount and retain the immunoactivity of recombinant dust mite allergen Der f2 immobilized on a glassy carbon electrode (GCE). The interaction between allergen and antibody was studied by electrochemical impedance spectroscopy (EIS). Self-assembled Au colloid layer (Phi = 16 nm) deposited on (3-mercaptopropyl)trimethoxysilane (MPTS)-modified GCE offered a basis to control the immobilization of allergen Der f2. The impedance measurements were based on the charge transfer kinetics of the [Fe(CN)(6)](3-/4-) redox pair, compared with bare GCE, the immobilization of allergen Der f2 and the allergen-antibody interaction that occurred on the electrode surface altered the interfacial electron transfer resistance and thereby slowed down the charge transfer kinetics by reducing the active area of the electrode or by preventing the redox species in electrolyte solution from approaching the electrode. The interactions of allergen with various concentrations of monoclonal antibody were also monitored through the change of impedance response. The results showed that the electron transfer resistance increased with increasing concentrations of monoclonal antibody.

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The ordered double perovskites, Sr2-xLaxMnMoO6, were prepared by sol-gel reaction. Structural, magnetic, and electrical properties were investigated for a series of ordered double perovskites Sr2-xLaxMnMoO6(0 <= x <= 1). The compounds have a monoclinic structure (space group P2(1)/n) and the cell volume expands monotonically with La doping. The T-C and the magnetic moment rise and the cusp-like transition temperature below which the magnetic frustration occurs shifts to high temperature as x increases. With La doping, electrical resistivity of Sr2-xLaxMnMoO6 decreases only at low doping levels (x <= 0.2); while at high doping levels (0.8 <= x <= 1), electrical resistivity tends to increase greatly. The results suggest that the competition between band filling effect and steric effect coexists in the whole doping range, and the formation of ferrimagnetic interactions is not simply at the expense of antiferromagnetic interactions.

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A novel sensitive electrochemical immunoassay with colloidal gold as the antibody labeling tag and subsequent signal amplification by silver enhancement is described. Colloidal gold was treated by a light-sensitive silver enhancement system which made silver deposit on the surface of colloidal gold(form Au/Ag core-shell structure), followed by the release of the metallic silver atoms anchored on the antibody by oxidative dissolution of them in an acidic solution and the indirect determination of the dissolved Ag+ ions by anodic stripping voltammetry(ASV) at a carbon fiber microelectrode. The electrochemical signal is directly proportional to the amount of analyte(goat IgG) in the standard or a sample. The method was evaluated by means of a noncompetitive heterogeneous immunoassay of immunoglobulin G(IgG) with a concentration as low as 0.2 ng/ mL. The high performance of the method is related to the sensitive ASV determination of silver(I) at a carbon fiber microelectrode and to the release of a large number of Ag+ ions from each silver shell anchored on the analyte(goat IgG).

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Protein multilayers composed of avidin and biotin-labeled antibody (bio-Ab) were prepared on gold surface by layer-by-layer assembly technology using the high specific binding constant (K-a: approximate to 10(15) M-1) between avidin and biotin. The assembly process of the multilayer films was monitored by using real-time BIA technique based on surface plasmon resonance (SPR). The multilayer films were also characterized by electrochemical impedance spectroscopy (EIS) and reflection absorption Fourier transform infrared spectroscopy (FTIR). The results indicate that the growth of the multilayer is uniform. From response of SPR for each layer, the stoichiometry S for the interaction between avidin and bio-Ab is calculated to be 0.37 in the multilayer whereas 0.82 in the first layer. The protein mass concentration for each layer was also obtained. The schematic figure for the multilayer assembly was proposed according to the layer mass, concentration and S value. The utility of the mutilayer films for immunosensing has been investigated via their subsequent interaction with hIgG. The binding ability of the multilayer increased for one to three layers of antibody, and then reach saturation after the fourth layer. These layer-by-layer constructed antibody multilayers enhance the binding ability than covalently immobilized monolayer antibody. This technology can be also used for construction of other thin films for immunosensing and biosensor.