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61.
AC voltammetric carbon paste-based enzyme immunosensors 总被引:10,自引:0,他引:10
Fernández-Sánchez C González-García MB Costa-García A 《Biosensors & bioelectronics》2000,14(12):917-924
Carbon paste electrodes, previously anodised in a basic media, are the basis for the development of a new voltammetric immunosensor device. Passive adsorption of the appropriate immunochemical reagent was performed onto the electrode surface. Alkaline Phosphatase labelled immunoglobulin was the tracer used in this work, 3-indoxyl phosphate being a very suitable enzymatic substrate for the electrochemical detection of the corresponding affinity reaction. The hydrolysis of this molecule generates indigo dimmer. This product was detected by alternating current voltammetry taking advantage of the adsorptive and inherent electrodic properties that it exhibits. The same electrochemical anodisation was used at the end of one assay to remove the entire protein layer attached to the carbon paste surface, allowing the formation of a new sensing phase and the use of the same support in several consecutive experiments. The methodology was applied to the design of two different immunoassays for the determination of human IgG. Good reproducibility of the electrodic signal and a limit of detection around 10−10 M were achieved. 相似文献
62.
Paddy Kane Kevin Kincaid Darren Fayne Dermot Diamond M. Anthony McKervey 《Journal of molecular modeling》2000,6(2):272-281
This paper focuses on the molecular modelling of a number of calixarene ester and phosphine oxide metal ion complexes. Monte Carlo conformational searches, in conjunction with the Merck Molecular Force Field, were carried out using Spartan SGI Version 5.0.1. running on Silicon Graphics O2 workstations. In the case of the calix[4]arene tetraesters, the optimised models strongly suggest that the selectivity of these ligands is strongly related to the eight-fold nature of the coordination with the Na+ ion, while coordination with the Li+ ion, for example, is merely three-fold. This feature of eight-fold coordination is also observed in the models of the complexes formed by the calix[4]arene tetraphosphine oxides with calcium. However, whereas the eight-fold coordination is unique to the model of the TPOL:Ca2+ complex among the ions modelled, this mode of coordination occurs for TPOS with sodium and potassium, in addition to calcium. This concurs with the observation that calcium selectivity is obtained with ion selective electrodes based on TPOL but not TPOS. Though the cavity in the calix[5]arenes PPOL and PPOLx and the calix[6]arene HPOL, in their uncomplexed form, are much larger than that of the corresponding calix[4]arenes, the pattern of selectivity is the same – the ligands are selective for calcium. The models of the complexes of these larger calixarenes, such as PPOL:Ca2+, strongly suggest that the reason for this similarity is that four of the available phosphine oxide groups complex with the calcium ion, and the others are forced away from the cavity region for steric reasons. The resulting eight-fold coordination, is therefore, similar to that of the calix[4]arenes studied.Electronic Supplementary Material available. 相似文献
63.
Abstract: The effects of substrates m -tyramine and β-phenethylamine, as well as cocaine, on the DA efflux from a cell line stably expressing the human norepinephrine transporter (hNET) were investigated by using rotating disk electrode voltammetry. Both the substrates and cocaine induced apparent DA efflux in a concentration-dependent manner. Their EC50 values for inducing DA efflux were similar to their IC50 values for inhibiting DA uptake. The substrate-induced DA efflux was inhibited by various NET blockers, enhanced by raising the internal [Na+ ] with Na+ ,K+ -ATPase inhibition, but was insensitive to membrane potential-altering agents valinomycin, veratridine, and high [K+ ]. The initial rate of m -tyramine-induced DA efflux was related to preloaded [DA] in a manner defined by a Michaelis-Menten expression. In contrast, DA efflux in the presence of cocaine displayed a much slower efflux rate, lower efficacy, was not stimulated by elevated internal [Na+ ], and was nonsaturable with preloaded [DA]. Single exponential kinetic analysis of the entire time course of the DA efflux showed that the apparent first-order rate constant for m -tyramine-induced DA efflux declined with increased preloaded [DA], whereas that for the DA efflux in the presence of cocaine was unchanged with varying preloaded [DA]. These results suggest that the substrates stimulate the NET-dependent DA efflux by increasing the accessibility of the NET to internal DA, whereas cocaine "uncovers" NET-independent DA efflux by reducing the accessibility of diffused/leaked external DA to the NET. 相似文献
64.
We measured the light response curve of photosynthetic oxygen evolution by illuminating a leaf disc in an air-tight windowed chamber. Oxygen production was measured by monitoring the quenching of luminescence of an organometallic ruthenium compound. A photodiode based chlorophyll a fluorometer was used to measure the luminescence intensity. Oxygen evolution measurements with a traditional oxygen electrode gave the same numerical values at different light intensities when the same leaf disk was tested. The quality of the measurement signal of the new method was found to be similar to that obtained with the oxygen electrode method. The new luminescence based system is more stable against electrical disturbances than an oxygen electrode, its response to oxygen pressure changes is very rapid, and the new method allows the same basic equipment to be used for chlorophyll fluorescence and oxygen measurements. 相似文献
65.
随着可穿戴式健康监测技术的发展,新型心电传感器-织物电极成为人们关注的热点,本文对织物电极的皮肤-电极接触阻抗测量方法进行了综述。首先介绍了织物电极的概念,分析了织物电极的皮肤-电极电化学界面、皮肤-电极电化学界面的等效电路和简化电路模型,得出了皮肤-电极接触阻抗的计算公式;其次,将皮肤-电极接触阻抗的测量方法归纳为直接测量法、参比测量法和模拟皮肤测量法三类,讨论了它们的测量原理和优缺点。本文认为需将模拟皮肤测量法和真实皮肤测量法有机结合,才能有效评价织物电极的阻抗特性,为织物电极的性能评价和心电信号采集电路的设计提供重要依据。最后,本文对织物电极待解决的问题进行了分析讨论。 相似文献
66.
Quantum-chemical study of structures, energies, and effective partial charge distribution for several models of the Rieske protein redox center is performed in terms of the B3LYP density functional method in combination with the broken symmetry approach using three different atomic basis sets. The structure of the redox complex optimized in vacuum differs markedly from that inside the protein. This means that the protein matrix imposes some stress on the active site resulting in distortion of its structure. The redox potentials calculated for the real active site structure are in a substantially better agreement with the experiment than those calculated for the idealized structure. This shows an important role of the active site distortion in tuning its redox potential. The reference absolute electrode potential of the standard hydrogen electrode is used that accounts for the correction caused by the water surface potential. Electrostatic calculations are performed in the framework of the polarizable solute model. Two dielectric permittivities of the protein are employed: the optical permittivity for calculation of the intraprotein electric field, and the static permittivity for calculation of the dielectric response energy. Only this approach results in a reasonable agreement of the calculated and experimental redox potentials. 相似文献
67.
The impact of mineral N supply, N-free or NO3(-) with or without NH4+, on the subsequent uptake of NO3(-) by maritime pine seedlings associated with the ectomycorrhizal fungus Rhizopogon roseolus was studied using ion-selective microelectrodes. NO3(-) net fluxes into N-starved non-mycorrhizal short roots (NMSRs) were low and measurable only over the NO3(-) concentration range of 0-70 microM. The simple kinetics observed in those roots may reflect the dominant operation of a high-affinity NO3(-) transport system (HATS) which is constitutive. NO3(-) pretreatment increased the NO3(-) net fluxes and led to a complex kinetics that may reflect the operation of other HATS. A simple kinetics was observed in plants pre-incubated at high NH4+ concentration. In contrast, NO3(-) uptake kinetics presented only one saturation phase in the fungus, whether associated with the plant or not. NO3(-) uptake was greater after a pretreatment in N-free or NO3 (-) solution, but NH4+ pretreatment led to a threefold reduction in NO3 (-) uptake. These results suggest that the regulation of NO3(-) transport systems varies between the host and the fungal partner. This variation is likely to contribute to the positive effect of mycorrhizal association on N uptake in plants when the N supply is low and fluctuating. 相似文献
68.
Synthesis and characterization of metal ion imprinted nano-porous polymer for the selective recognition of copper 总被引:1,自引:0,他引:1
Baghel A Boopathi M Singh B Pandey P Mahato TH Gutch PK Sekhar K 《Biosensors & bioelectronics》2007,22(12):3326-3334
Selective recognition of metal ions utilizing metal ion-imprinted polymers (MIIPs) received much importance in diverse fields owing to their high selectivity for the target metal ions. In the present study, a copper ion imprinted polymer was synthesized without an additional complexing ligand or complex with a broad aim to avoid the conventional extra metal ion complexing ligand during the synthesis of MIIP. The complete removal of the copper metal ion from the MIIP was confirmed by AAS and SEM–EDX. SEM image of the MIIP exhibited nano-patterns and it was also found to be entirely different from that of non-imprinted polymer and polymer with copper metal ions. BET surface area analysis revealed more surface area (47.96 m2/g) for the Cu(II)-MIIP than non-imprinted control polymer (41.43 m2/g). TGA result of polymer with copper metal ion indicated more char yield (18.41%) when compared to non-imprinted control polymer (8.3%) and Cu(II)-MIIP (less than 1%). FTIR study confirmed the complexation between Cu(II)-MIIP and Cu(II) metal ion through carbonyl oxygen of acryl amide. The Cu(II)-MIIP exhibited an imprinting efficiency of 2.0 and it was showing 8% interference from a mixture of Zn, Ni and Co ions. A potentiometric ion selective electrode devised with Cu(II)-MIIP showed more potential response for Cu(II) ion than that was fabricated from non-imprinted polymer. 相似文献
69.
An electrochemical technique for the real-time detection of hydrogen peroxide (H2O2) was employed to describe respiratory burst activity (RBA) of phagocytes in plasma which can be used to evaluate the ability of immune system and disease resistance. The method is based upon the electric current changes, by redox reaction on platinum electrode of extracellular hydrogen peroxide (H2O2) released from phagocytes stimulated by the zymosan at 680 mV direct current (d.c.). Compared with the control, activation of respiratory burst by zymosan particles results in a high amperometric response, and a current peak was obtained during the whole monitoring process. The peak current was proved by addition of Cu2+ and other controls, to be the result of intense release of H2O2 from phagocytes. The peak area was calculated and used to evaluate the quantity of effective H2O2, which represents the quantity of H2O2 beyond the clearance of related enzymes in plasma. According to Faraday's law, the phagocytes' ability of prawns to generate effective H2O2 was evaluated from 1.253 x 10(-14) mol/cell to 6.146 x 10(-14) mol/cell, and carp from 1.689 x 10(-15) mol/cell to 7.873 x 10(-15) mol/cell. This method is an acute and quick detection of extracellular effective H2O2 in plasma and reflects the capacity of phagocytes under natural conditions, which could be applied for selecting species and parents with high immunity for breeding in aquaculture. 相似文献
70.
The interfacial electron transfer of glucose oxidase (GOx) on a poly(glutamic acid)-modified glassy carbon electrode (PGA/GCE) was investigated. The redox peaks measured for GOx and flavin adenine dinucleotide (FAD) are similar, and the anodic peak of GOx does not increase in the presence of glucose in a mediator-free solution. These indicate that the electroactivity of GOx is not the direct electron transfer (DET) between GOx and PGA/GCE and that the observed electroactivity of GOx is ascribed to free FAD that is released from GOx. However, efficient electron transfer occurred if an appropriate mediator was placed in solution, suggesting that GOx is active. The PGA/GCE-based biosensor showed wide linear response in the range of 0.5–5.5 mM with a low detection limit of 0.12 mM and high sensitivity and selectivity for measuring glucose. 相似文献