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This paper describes the construction and characterization of an electrochemical nitrate biosensor based on the ultrathin-film composite membrane concept. The composite membrane is prepared by electropolymerization of a thin anion-permselective coating of 1-methyl-3-(pyrrol-1-ylmethyl)pyridinium across the surface of a microporous support membrane. This film separates the analyte solution from an internal sensing solution which contains the enzyme nitrate reductase and an electrocatalyst (methyl viologen). The ultrathin anion-permselective fill prevents loss of these components from the internal sensing solution, yet allows the analyte, N[O.sub.3]-, to enter the internal solution. A glassy carbon working electrode is immersed into the internal solution. The nitrate is reduced enzymatically, yielding the oxidized form of the enzyme, which is reduced again by electrogenerated methyl viologen radical cation. The nitrate concentration in the analyte solution is proportional to the catalytic methyl viologen reduction current. This sensor shows good sensitivity to nitrate, with a detection limit of 5.4 [[micro]meter] and a dynamic range which extends up to 100 [[micro]meter] N[O.sub.3]-.

Ensembles of nanoscopic disk-shaped electrodes have been shown to offer enhancements in electroanalytical detection limits relative to electrodes of macroscopic dimensions (e.g., disk electrodes with diameters of [approximately]1 mm). Enhancements in electroanalytical detection limits have also been observed at macroscopic electrodes that have been coated with films of ion-exchange polymers. In this paper we combine these two concepts.. We demonstrate that a nanoelectrode ensemble (NEE) that has been coated with a thin film of the Kodak ion-exchange polymer AQ 55 shows enhanced electroanalytical detection limits relative to the uncoated NEE and to the coated macroscopic electrode. To our knowledge, this is the first investigation of the electrochemistry, and the electroanalytical advantages, of polymer film-coated NEEs.

Since the first reports dating back to the mid-1990s, ensembles and arrays of nanoelectrodes (NEEs and NEAs, respectively) have gained an important role as advanced electroanalytical tools thank to their unique characteristics which include, among others, dramatically improved signal/noise ratios, enhanced mass transport and suitability for extreme miniaturization. From the year 2000 onward, these properties have been exploited to develop electrochemical biosensors in which the surfaces of NEEs/NEAs have been functionalized with biorecognition layers using immobilization modes able to take the maximum advantage from the special morphology and composite nature of their surface. This paper presents an updated overview of this field. It consists of two parts. In the first, we discuss nanofabrication methods and the principles of functioning of NEEs/NEAs, focusing, in particular, on those features which are important for the development of highly sensitive and miniaturized biosensors. In the second part, we review literature references dealing the bioanalytical and biosensing applications of sensors based on biofunctionalized arrays/ensembles of nanoelectrodes, focusing our attention on the most recent advances, published in the last five years. The goal of this review is both to furnish fundamental knowledge to researchers starting their activity in this field and provide critical information on recent achievements which can stimulate new ideas for future developments to experienced scientists. [ABSTRACT FROM AUTHOR]

Immunoglobulin Y is a water-soluble protein present in high concentration in hen serum and egg yolk. IgY has applications in many fields, e.g., from food stuff to the mass production of antibodies. In this work, we have implemented an electrochemical immunosensor for IgY based on templated nanoelectrodes ensembles. IgY is captured by the templating polycarbonate and reacted with anti-IgY labeled with horseradish peroxidase. In the presence of H2O2 and methylene blue as the redox mediator, an electrocatalytic current is generated which scales with IgY concentration in the sample. After optimizing the extracting procedure, the immunosensor was applied for analysis of fresh eggs and food integrators. The data obtained with the biosensor were validated by SDS-PAGE and Western blot measurements. [ABSTRACT FROM AUTHOR]

Celiac disease (CD) is a gluten-dependent autoimmune disorder affecting a significant percentage of the general population, with increasing incidence particularly for children. Reliable analytical methods suitable for the serological diagnosis of the disorder are urgently required for performing both the early diagnosis and the follow-up of a patient adhering to a gluten-free diet. Herein we report on the preparation and application of a novel electrochemical immunosensor based on the use of ensembles of gold nanoelectrodes (NEEs) for the detection of anti-tissue transglutaminase (anti-tTG), which is considered one reliable serological marker for CD. To this end, we take advantage of the composite nature of the nanostructured surface of membrane-templated NEEs by functionalizing the polycarbonate surface of the track-etched membrane with tissue transglutaminase. Incubation of the functionalized NEE in anti-tTG samples results in the capture of the anti-tTG antibody. Confirmation of the recognition event is achieved by incubating the NEE with a secondary antibody labelled with horseradish peroxidase (HRP): in the presence of H2O2 as substrate and hydroquinone as redox mediator, an electrocatalytic current is indeed generated whose increment is proportional to the amount of anti-tTG captured from the sample. The optimized sensor allows a detection limit of 1.8 ng mL−1, with satisfactory selectivity and reproducibility. Analysis of serum samples from 28 individuals, some healthy and some affected by CD, furnished analytical results comparable with those achieved by classical fluoroenzyme immunoassay (FEIA). We note that the NEE-based immunosensor developed here detects the IgG isotype of anti-tTG, while FEIA detects the IgA isotype, which is not a suitable diagnostic marker for IgA-deficient patients. [ABSTRACT FROM AUTHOR]

This work is aimed at developing an electrochemical sensor for the sensitive and selective detection of trace levels of perfluorooctanesulfonate (PFOS) in water. Contamination of waters by perfluorinated alkyl substances (PFAS) is a problem of global concern due to their suspected toxicity and ability to bioaccumulate. PFOS is the perfluorinated compound of major concern, as it has the lowest suggested control concentrations. The sensor reported here is based on a gold electrode modified with a thin coating of a molecularly imprinted polymer (MIP), prepared by anodic electropolymerization of o-phenylenediamine (o-PD) in the presence of PFOS as the template. Activation of the sensor is achieved by template removal with suitable a solvent mixture. Voltammetry, a quartz crystal microbalance, scanning electron microscopy and elemental analysis were used to monitor the electropolymerization process, template removal, and binding of the analyte. Ferrocenecarboxylic acid (FcCOOH) has been exploited as an electrochemical probe able to generate analytically useful voltammetric signals by competing for the binding sites with PFOS, as the latter is not electroactive. The sensor has a low detection limit (0.04 nM), a satisfactory selectivity, and is reproducible and repeatable, giving analytical results in good agreement with those obtained by HPLC-MS/MS analyses.