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1.
Int J Mol Sci ; 23(16)2022 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-36012396

RESUMO

Current procedures for the assessment of chronic wound infection are time-consuming and require complex instruments and trained personnel. The incidence of chronic wounds worldwide, and the associated economic burden, urge for simple and cheap point-of-care testing (PoCT) devices for fast on-site diagnosis to enable appropriate early treatment. The enzyme myeloperoxidase (MPO), whose activity in infected wounds is about ten times higher than in non-infected wounds, appears to be a suitable biomarker for wound infection diagnosis. Herein, we develop a single-component foldable paper-based device for the detection of MPO in wound fluids. The analyte detection is achieved in two steps: (i) selective immunocapture of MPO, and (ii) reaction of a specific dye with the captured MPO, yielding a purple color with increasing intensity as a function of the MPO activity in infected wounds in the range of 20-85 U/mL. Ex vivo experiments with wound fluids validated the analytic efficiency of the paper-based device, and the results strongly correlate with a spectrophotometric assay.


Assuntos
Líquidos Corporais , Infecção dos Ferimentos , Colorimetria , Corantes , Humanos , Papel , Testes Imediatos , Infecção dos Ferimentos/diagnóstico
2.
Int J Mol Sci ; 23(23)2022 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-36499587

RESUMO

SARS-CoV-2, a positive-strand RNA virus has caused devastating effects. The standard method for COVID diagnosis is based on polymerase chain reaction (PCR). The method needs expensive reagents and equipment and well-trained personnel and takes a few hours to be completed. The search for faster solutions has led to the development of immunological assays based on antibodies that recognize the viral proteins that are faster and do not require any special equipment. Here, we explore an innovative analytical approach based on the sandwich oligonucleotide hybridization which can be adapted to several biosensing devices including thermal lateral flow and electrochemical devices, as well as fluorescent microarrays. Polypurine reverse-Hoogsteen hairpins (PPRHs) oligonucleotides that form high-affinity triplexes with the polypyrimidine target sequences are used for the efficient capture of the viral genome. Then, a second labeled oligonucleotide is used to detect the formation of a trimolecular complex in a similar way to antigen tests. The reached limit of detection is around 0.01 nM (a few femtomoles) without the use of any amplification steps. The triplex enhanced nucleic acid detection assay (TENADA) can be readily adapted for the detection of any pathogen requiring only the knowledge of the pathogen genome sequence.


Assuntos
COVID-19 , Ácidos Nucleicos , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , Oligonucleotídeos/química , Reação em Cadeia da Polimerase , RNA Viral/genética , RNA Viral/análise , Técnicas de Amplificação de Ácido Nucleico/métodos
3.
Sensors (Basel) ; 20(4)2020 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-32054035

RESUMO

In healthcare, new diagnostic tools that help in the diagnosis, prognosis, and monitoring of diseases rapidly and accurately are in high demand. For in-situ measurement of disease or infection biomarkers, point-of-care devices provide a dramatic speed advantage over conventional techniques, thus aiding clinicians in decision-making. During the last decade, paper-based analytical devices, combining paper substrates and electrochemical detection components, have emerged as important point-of-need diagnostic tools. This review highlights significant works on this topic over the last five years, from 2015 to 2019. The most relevant articles published in 2018 and 2019 are examined in detail, focusing on device fabrication techniques and materials applied to the production of paper fluidic and electrochemical cell architectures as well as on the final device assembly. Two main approaches were identified, that are, on one hand, those ones where the fabrication of the electrochemical cell is done on the paper substrate, where the fluidic structures are also defined, and, on the other hand, the fabrication of those ones where the electrochemical cell and liquid-driving paper component are defined on different substrates and then heterogeneously assembled. The main limitations of the current technologies are outlined and an outlook on the current technology status and future prospects is given.


Assuntos
Biomarcadores/análise , Técnicas Biossensoriais/métodos , Papel , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Técnicas Eletroquímicas , Humanos , Sistemas Automatizados de Assistência Junto ao Leito
4.
Int J Mol Sci ; 21(16)2020 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-32796622

RESUMO

Ascorbate oxidases are an enzyme group that has not been explored to a large extent. So far, mainly ascorbate oxidases from plants and only a few from fungi have been described. Although ascorbate oxidases belong to the well-studied enzyme family of multi-copper oxidases, their function is still unclear. In this study, Af_AO1, an enzyme from the fungus Aspergillus flavus, was characterized. Sequence analyses and copper content determination demonstrated Af_AO1 to belong to the multi-copper oxidase family. Biochemical characterization and 3D-modeling revealed a similarity to ascorbate oxidases, but also to laccases. Af_AO1 had a 10-fold higher affinity to ascorbic acid (KM = 0.16 ± 0.03 mM) than to ABTS (KM = 1.89 ± 0.12 mM). Furthermore, the best fitting 3D-model was based on the ascorbate oxidase from Cucurbita pepo var. melopepo. The laccase-like activity of Af_AO1 on ABTS (Vmax = 11.56 ± 0.15 µM/min/mg) was, however, not negligible. On the other hand, other typical laccase substrates, such as syringaldezine and guaiacol, were not oxidized by Af_AO1. According to the biochemical and structural characterization, Af_AO1 was classified as ascorbate oxidase with unusual, laccase-like activity.


Assuntos
Ascorbato Oxidase/metabolismo , Aspergillus flavus/enzimologia , Lacase/metabolismo , Sequência de Aminoácidos , Ascorbato Oxidase/química , Cobre/metabolismo , Cinética , Lacase/química , Modelos Moleculares , Oxirredução , Especificidade por Substrato
5.
Anal Chem ; 91(24): 15959-15966, 2019 12 17.
Artigo em Inglês | MEDLINE | ID: mdl-31750653

RESUMO

Performance of electroanalytical lab-on-a-chip devices is often limited by the mass transfer of electroactive species toward the electrode surface, due to the difficulty in applying external convection. This article describes the powerful signal enhancement attained with a 2.54 GHz miniature acoustic resonator integrated with an electrochemical device in a miniaturized cell. Acoustic resonator and an on-chip gold thin-film three-electrode electrochemical cell were arranged facing each other inside a structured poly(methyl methacrylate) chamber. Cyclic voltammetric and chronoamperometric responses of 1 mM ferrocene-methanol were recorded under resonator's actuation at powers ranging from 0 to 1 W. Finite element analysis was carried out to study the sono-electroanalytical process. Acoustic resonator's actuation greatly enhances the mass transport of electroactive species toward the electrode surface. The diffusion limited cyclic voltammetric and chronoamperometric currents increase around 10 and 20 times, respectively, with an input power of 1 W compared to those recorded under stagnant conditions. The improvement in electroanalytical process is mainly associated with acoustic resonator's vibration induced fluid streaming. The advantages of a miniaturized acoustic resonator, including the submillimeter small size, amenability for mass fabrication, cost effectiveness, low energy consumption, as well as outstanding enhancement of coupled electrochemical processes, will enable the production of highly sensitive compact electroanalytical devices.

6.
Chem Rec ; 18(7-8): 749-758, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29806230

RESUMO

The synthesis of organic-inorganic nanocomposites that can interact with different environmental pollutants and can be mass-produced are very promising materials for the fabrication of chemical sensor devices. Among them, metal (or metal oxide) nanoparticles doped conductive porous carbon composites can be readily applied to the production of electrochemical sensors and show enhanced sensitivity for the measurement of water pollutants, thanks to the abundant accessible and functional sites provided by the interconnected porosity and the metallic nanoparticles, respectively. In this personal account, an overview of several synthesis routes of porous carbon composites containing metallic nanoparticles is given, paying special attention to those based on sol-gel techniques. These are very powerful to synthesize hybrid porous materials that can be easily processed into powders and thin films, so that they can be implemented in electrode fabrication processes based on screen-printing and lithography techniques, respectively. We emphasize the sol-gel routes developed in our group for the synthesis of bismuth or gold nanoparticle doped porous carbon composites applied to fabricate electrochemical sensors that can be scaled down to produce miniaturized on-chip sensing devices for the sensitive detection of heavy metal pollutants in water. The trend towards the miniaturization of electrochemical sensors to be readily employed as analytical tools in environmental monitoring follow the market requirements of rapid and accurate on-site analysis, small sample consumption and waste production, as well as potential for continuous or semi-continuous in-situ determination of a wide variety of target analytes.

7.
Anal Chem ; 87(4): 2152-60, 2015 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-25594378

RESUMO

This work reports on the fabrication and comparative analytical assessment of electrochemical sensors applied to the rapid analysis of chemical oxygen demand (COD) in urban waste waters. These devices incorporate a carbon nanotube-polystyrene composite, containing different inorganic electrocatalysts, namely, Ni, NiCu alloy, CoO, and CuO/AgO nanoparticles. The sensor responses were initially evaluated using glucose as standard analyte and then by analyzing a set of real samples from urban wastewater treatment plants. The estimated COD values in the samples were compared with those provided by an accredited laboratory using the standard dichromate method. The sensor prepared with the CuO/AgO-based nanocomposite showed the best analytical performance. The recorded COD values of both the sensor and the standard method were overlapped, considering the 95% confidence intervals. In order to show the feasible application of this approach for the detection of COD online and in continuous mode, the CuO/AgO-based nanocomposite sensor was integrated in a compact flow system and applied to the detection of wastewater samples, showing again a good agreement with the values provided by the dichromate method.

8.
Biosens Bioelectron ; 262: 116560, 2024 Jul 11.
Artigo em Inglês | MEDLINE | ID: mdl-39018979

RESUMO

The development of wearable devices for sweat analysis has experienced significant growth in the last two decades, being the main focus the monitoring of athletes health during workouts. One of the main challenges of these approaches has been to attain the continuous monitoring of sweat for time periods over 1 h. This is the main challenge addressed in this work by designing an analytical platform that combines the high performance of potentiometric sensors and a fluidic structure made of a plastic fabric into a multiplexed wearable device. The platform comprises Ion-Sensitive Field-Effect Transistors (ISFETs) manufactured on silicon, a tailor-made solid-state reference electrode, and a temperature sensor integrated into a patch-like polymeric substrate, together with the component that easily collects and drives samples under continuous capillary flow to the sensor areas. ISFET sensors for measuring pH, sodium, and potassium ions were fully characterized in artificial sweat solutions, providing reproducible and stable responses. Then, the real-time and continuous monitoring of the biomarkers in sweat with the wearable platform was assessed by comparing the ISFETs responses recorded during an 85-min continuous exercise session with the concentration values measured using commercial Ion-Selective Electrodes (ISEs) in samples collected at certain times during the session. The developed sensing platform enables the continuous monitoring of biomarkers and facilitates the study of the effects of various real working conditions, such as cycling power and skin temperature, on the target biomarker concentration levels.

9.
Analyst ; 138(3): 839-44, 2013 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-23237871

RESUMO

The selective absorbance detection of mercury(II) (Hg(2+)) and lead(II) (Pb(2+)) ions using ferrocene-based colorimetric ligands and miniaturized multiple internal reflection (MIR) systems implemented in a low-cost photonic lab on a chip (PhLoC) is reported. The detection principle is based on the formation of selective stable complexes between the heavy metal ion and the corresponding ligand. This interaction modulates the ligand spectrum by giving rise to new absorbance bands or wavelength shifting of the existing ones. A comparative study for the detection of Hg(2+) was carried out with two MIR-based PhLoC systems showing optical path lengths (OPLs) of 0.64 cm and 1.42 cm as well as a standard cuvette (1.00 cm OPL). Acetonitrile solutions containing the corresponding ligand and increasing concentrations of the heavy metal ion were pumped inside the systems and the absorbance in the visible region of the spectra was recorded. The optical behaviour of all the tested systems followed the expected Beer-Lambert law. Thus, the best results were achieved with the one with the longest OPL, which showed a linear behaviour in a concentration range of 1 µM-90 µM Hg(2+), a sensitivity of 5.6 × 10(-3) A.U. µM(-1) and a LOD of 2.59 µM (0.49 ppm), this being 1.7 times lower than that recorded with a standard cuvette, and using a sample/reagent volume around 190 times smaller. This microsystem was also applied for the detection of Pb(2+) and a linear behaviour in a concentration range of 3-100 µM was obtained, and a sensitivity of 9.59 × 10(-4) A.U. µM(-1) and a LOD of 4.19 µM (0.868 ppm) were achieved. Such a simple analytical tool could be implemented in portable instruments for automatic in-field measurements and, considering the minute sample and reagent volume required, would enable the deployment of high throughput environmental analysis of these pollutants and other related hazardous species.


Assuntos
Dimetilpolisiloxanos/química , Poluentes Ambientais/análise , Metais Pesados/análise , Espectrofotometria Ultravioleta , Compostos Ferrosos/química , Íons/química , Dispositivos Lab-On-A-Chip , Chumbo/análise , Mercúrio/análise , Metalocenos , Fótons , Espectrofotometria Ultravioleta/instrumentação
10.
ACS Sens ; 8(8): 3032-3042, 2023 08 25.
Artigo em Inglês | MEDLINE | ID: mdl-37467113

RESUMO

Health initiatives worldwide demand affordable point-of-care devices to aid in the reduction of morbidity and mortality rates of high-incidence infectious and noncommunicable diseases. However, the production of robust and reliable easy-to-use diagnostic platforms showing the ability to quantitatively measure several biomarkers in physiological fluids and that could in turn be decentralized to reach any relevant environment remains a challenge. Here, we show the particular combination of paper-microfluidic technology, electrochemical transduction, and magnetic nanoparticle-based immunoassay approaches to produce a unique, compact, and easily deployable multiplex device to simultaneously measure interleukin-8, tumor necrosis factor-α, and myeloperoxidase biomarkers in sputum, developed with the aim of facilitating the timely detection of acute exacerbations of chronic obstructive pulmonary disease. The device incorporates an on-chip electrochemical cell array and a multichannel paper component, engineered to be easily aligned into a polymeric cartridge and exchanged if necessary. Calibration curves at clinically relevant biomarker concentration ranges are produced in buffer and artificial sputum. The analysis of sputum samples of healthy individuals and acutely exacerbated patients produces statistically significant biomarker concentration differences between the two studied groups. The device can be mass-produced at a low cost, being an easily adaptable platform for measuring other disease-related target biomarkers.


Assuntos
Microfluídica , Nanopartículas , Humanos , Escarro , Sistemas Automatizados de Assistência Junto ao Leito , Biomarcadores/análise
11.
IEEE Trans Biomed Circuits Syst ; 17(4): 808-817, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37318976

RESUMO

Sweat secreted by the human eccrine sweat glands can provide valuable biomarker information during exercise. Real-time non-invasive biomarker recordings are therefore useful for evaluating the physiological conditions of an athlete such as their hydration status during endurance exercise. This work describes a wearable sweat biomonitoring patch incorporating printed electrochemical sensors into a plastic microfluidic sweat collector and data analysis that shows the real-time recorded sweat biomarkers can be used to predict a physiological biomarker. The system was placed on subjects carrying out an hour-long exercise session and results were compared to a wearable system using potentiometric robust silicon-based sensors and to commercially available HORIBA-LAQUAtwin devices. Both prototypes were applied to the real-time monitoring of sweat during cycling sessions and showed stable readings for around an hour. Analysis of the sweat biomarkers collected from the printed patch prototype shows that their real-time measurements correlate well (correlation coefficient ≥ 0.65) with other physiological biomarkers such as heart rate and regional sweat rate collected in the same session. We show for the first time, that the real-time sweat sodium and potassium concentration biomarker measurements from the printed sensors can be used to predict the core body temperature with root mean square error (RMSE) of 0.02 °C which is 71% lower compared to the use of only the physiological biomarkers. These results show that these wearable patch technologies are promising for real-time portable sweat monitoring analytical platforms, especially for athletes performing endurance exercise.


Assuntos
Técnicas Biossensoriais , Dispositivos Eletrônicos Vestíveis , Humanos , Suor/química , Temperatura Corporal , Eletrólitos , Biomarcadores/análise
12.
Anal Chem ; 84(8): 3546-53, 2012 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-22429156

RESUMO

A dual lab on a chip (DLOC) approach that enables simultaneous optical and electrochemical detection working in a continuous flow regime is presented. Both detection modes are integrated for the first time into a single detection volume and operate simultaneously with no evidence of cross-talk. The electrochemical cell was characterized amperometrically by measuring the current in ferrocyanide solutions at +0.4 V vs gold pseudoreference electrode, at a flow rate of 200 µL min(-1). The experimental results for ferrocyanide concentrations ranging from 0.005 to 2 mM were in good agreement with the values predicted by the Levich equation for a microelectrode inside a rectangular channel, with a sensitivity of 2.059 ± 0.004 µA mM(-1) and a limit of detection (LoD) of (2.303 ± 0.004) × 10(-3) mM. Besides, optical detection was evaluated by measuring the absorbance of ferricyanide solutions at 420 nm. The results obtained therein coincide with those predicted by the Beer-Lambert law for a range of ferricyanide concentrations from 0.005 to 0.3 mM and showed an estimated LoD of (0.553 ± 0.001) × 10(-3) mM. The DLOC was finally applied to the analysis of L-lactate via a bienzymatic reaction involving lactate oxidase (LOX) and horseradish peroxidase (HRP). Here, the consumption of the reagent of the reaction (ferrocyanide) was continuously monitored by amperometry whereas the product of the reaction (ferricyanide) was recorded by absorbance. The DLOC presented good performance in terms of sensitivity and limit of detection, comparable to other fluidic systems found in the literature. Additionally, the ability to simultaneously quantify enzymatic reagent consumption and product generation confers the DLOC a self-verifying capability which in turn enhances its robustness and reliability.


Assuntos
Eletroquímica/instrumentação , Eletroquímica/métodos , Ferrocianetos/química , Técnicas Analíticas Microfluídicas/instrumentação , Eletrodos , Ouro/química , Limite de Detecção , Soluções
13.
Opt Express ; 20(21): 23700-19, 2012 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-23188336

RESUMO

This work reports on the implementation of different absorption micro-filters based on a dye-doped hybrid organic-inorganic xerogel polymeric material synthesized by the sol-gel process. Microstructures containing eight different filter widths were fabricated in polydimethylsiloxane (PDMS), bonded to glass substrates and filled with the corresponding dye doped polymeric material by a soft lithography approach. The filtering capacity as a function of dye concentration and filter width was studied and revealed a linear dependence with both parameters, as expected according to the Beer-Lambert law. Zero passband transmittance values and relatively sharp stopband regions were achieved with all the filters, also showing rejection levels between -6 dB and -55 dB. Finally, such filters were monolithically integrated into a disposable fluorescence-based photonic lab-on-a-chip (PhLoC) approach. Calibration curves carried out with a model fluorophore target analyte showed an over two-fold increase in sensitivity and a thirty-fold decrease of the limit of detection (LOD) compared with the values recorded using the same PhLoC system but without the polymeric filter structure. The results presented herein clearly indicate the feasibility of these xerogel-based absorbance filtering structures for being applied as low-cost optical components that can be easily incorporated into disposable fluorescence-based photonic lab on a chip systems.


Assuntos
Dimetilpolisiloxanos/química , Filtração/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Nylons/química , Espectrometria de Fluorescência/instrumentação , Equipamentos Descartáveis , Desenho de Equipamento , Análise de Falha de Equipamento , Luz , Espalhamento de Radiação , Integração de Sistemas
14.
ACS Appl Mater Interfaces ; 14(35): 40182-40190, 2022 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-35998366

RESUMO

Bread waste is a major part of food wastage which could be upcycled to produce functional materials, following the principles of the circular bioeconomy. This work shows that bread waste can be recycled and valorized to produce a composite conductive material with excellent properties for chemical sensor applications. Here, dry bread is impregnated with an aqueous solution of a silver precursor and pyrolyzed to produce a porous carbon matrix containing Ag nanoparticles with diameters ranging from 20 to 40 nm. These particles perform as catalytic redox centers for the electrochemical detection of halide ions (Cl-, Br-, and I-) and organohalide target molecules such as sucralose and trichloroacetic acid. A thorough analytical characterization is carried out to show the potential application of the developed material for the manufacturing of electrochemical sensor approaches. The material preparation is sustainable, low-cost, simple, and upscalable. These are ideal features for the large-scale manufacturing by screen-printing technologies of single-use electrochemical sensors for the rapid analysis of halogenated organic pollutants in waters.

15.
Biosens Bioelectron ; 201: 113952, 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-34999523

RESUMO

This work reports on the fabrication and performance of a new on-chip array of gold thin-film electrodes arranged into five individually addressable miniaturized electrochemical cells. Each cell shows a two-electrode configuration comprising a single working electrode and a counter/pseudo-reference electrode that is compartmentalized to be shared among all the cells of the array. Using this configuration, just six contact pads are required, which significantly reduces the chip overall surface area. Electrochemical characterization studies are carried out in solutions containing the two species of reversible redox pairs. The concentration of one redox species can reliably be measured at the working electrode by applying potentiostatic techniques to record the current due to the corresponding electrochemical reaction. The redox counterpart in turn undergoes an electrochemical process at the counter/pseudo-reference electrode, which, under optimized experimental conditions, injects current and keeps the applied potential in the electrochemical cell without limiting the current being recorded at the working electrode. Under these conditions, the electrode array shows an excellent performance in electrochemical detection studies without any chemical or electrical cross-talk between cells. The enzymatic activity of horseradish peroxidase, alkaline phosphatase and myeloperoxidase enzymes is analyzed using different redox mediators. Quasi-simultaneous measurements with the five electrochemical cells of the array are carried out within 1 s time frame. This array layout can be suitable for multiplexed electrochemical immunoassays and immunosensor approaches and implementation in simplified electrochemical ELISA platforms that make use of enzyme labels. Moreover, the array reduced dimensions facilitate the integration into compact fluidic devices.


Assuntos
Técnicas Biossensoriais , Técnicas Eletroquímicas , Eletrodos , Ouro , Peroxidase do Rábano Silvestre , Imunoensaio
16.
Biosensors (Basel) ; 12(5)2022 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-35624581

RESUMO

Lab-on-a-chip devices incorporating valves and pumps can perform complex assays involving multiple reagents. However, the instruments used to drive these chips are complex and bulky. In this article, a new wax valve design that uses light from a light emitting diode (LED) for both opening and closing is reported. The valves and a pumping chamber are integrated in lab-on-a-foil chips that can be fabricated at low cost using rapid prototyping techniques. A chip for the implementation of enzyme-linked immunosorbent assays (ELISA) is designed. A porous nitrocellulose material is used for the immobilization of capture antibodies in the microchannel. A compact generic instrument with an array of 64 LEDs, a linear actuator to drive the pumping chamber, and absorbance detection for a colorimetric readout of the assay is also presented. Characterization of all the components and functionalities of the platform and the designed chip demonstrate their potential for assay automation.


Assuntos
Dispositivos Lab-On-A-Chip , Microfluídica , Automação , Colorimetria , Ensaio de Imunoadsorção Enzimática
17.
Anal Chem ; 83(5): 1726-31, 2011 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-21288019

RESUMO

A new approach for the electrical readout of microarrays prepared on regular glass slides, using an array of impedimetric transducers (interdigitated electrodes, IDEs) is presented in this work. Impedance detection relies on the use of a urease-labeled immunoassay scheme. Urease is able to produce an increase in conductivity by hydrolysis of the urea substrate, which is measured with the IDEs and directly related to the amount of target analyte. Unlike previous electrical microarrays, the assay does not take place on top of the transducers but on a regular glass slide, which may enable the development of compact multiplexed analytical systems with lower cost per assay. A droplet of solution with the enzymatic substrate is deposited on each transducer of the array, and the microarray is positioned at a short distance (300 µm) so that each droplet wets one transducer and one spot of the microarray. This procedure allows reusing the transducer array for readout of a virtually unlimited number of microarrays. A microarray based on an immunoassay for the detection of a mouse generic protein in a concentration range from 0.03 to 30 µg mL(-1) was carried out to assess the performance of the electrical readout approach. A sigmoid response with a limit of detection of 0.1 µg mL(-1) and a dynamic range of 1 order of magnitude was obtained. A comparative study was also carried out with two well established analytical procedures. First, the urease-based immunoassay was tested in a 96 well microtiter plate using phenol red pH indicator and absorbance detection. Second, the microarray was carried out using the same target protein concentration range but applying a Cy3 label and fluorescence detection. Both assays allowed for the validation of the performance of the presented electrical readout system.


Assuntos
Vidro , Análise Serial de Proteínas , Animais , Eletricidade , Camundongos
18.
Analyst ; 136(17): 3496-502, 2011 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-21336349

RESUMO

A comparative study of different approaches for the selective immobilisation of biomolecules on the surface of poly(dimethylsiloxane) (PDMS) is reported. The motivation of this work is to set a robust and reliable protocol for the easy implementation of a biosensor device in a PDMS-based photonic lab-on-a-chip (PhLoC). A hollow prism configuration, previously reported for the colorimetric detection of analytes was chosen for this study. Here, the inner walls of the hollow prism were initially modified by direct adsorption of either polyethylene glycol (PEG) or polyvinyl alcohol (PVA) linear polymers as well as by carrying out a light chemical oxidation step. All these processes introduced hydroxyl groups on the PDMS surface to a different extent. The hydroxyl groups were further silanised using a silane containing an aldehyde end-group. The interaction between this group and a primary amine moiety enabled the selective covalent attachment of a biomolecule on the PDMS surface. A thorough structural characterisation of the resulting modified-PDMS substrates was carried out by contact angle measurements, X-ray photoelectron spectroscopic (XPS) analysis and atomic force microscopy (AFM) imaging. Using horseradish peroxidase as a model recognition element, different biosensor approaches based on each modification process were developed for the detection of hydrogen peroxide target analyte in a concentration range from 0.1 µM to 100 µM. The analytical performance was similar in all cases, a linear concentration range between 0.1 µM and 24.2 µM, a sensitivity of 0.02 a.u. µM(-1) and a limit of detection around 0.1 µM were achieved. However, important differences were observed in the reproducibility of the devices as well as in their operational stability, which was studied over a period of up to two months. Considering all these studies, the PVA-modified approach appeared to be the most suitable one for the simple fabrication of a biosensor device integrated in a PDMS PhLoC.


Assuntos
Técnicas Biossensoriais/métodos , Dimetilpolisiloxanos/química , Peróxido de Hidrogênio/análise , Técnicas Analíticas Microfluídicas/métodos , Técnicas Biossensoriais/instrumentação , Enzimas Imobilizadas/metabolismo , Desenho de Equipamento , Peroxidase do Rábano Silvestre/metabolismo , Técnicas Analíticas Microfluídicas/instrumentação , Polietilenoglicóis/química , Álcool de Polivinil/química , Sensibilidade e Especificidade , Propriedades de Superfície
19.
Biosensors (Basel) ; 11(9)2021 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-34562893

RESUMO

The development of diagnostic tools for measuring a wide spectrum of target analytes, from biomarkers to other biochemical parameters in biological fluids, has experienced a significant growth in the last decades, with a good number of such tools entering the market. Recently, a clear focus has been put on miniaturized wearable devices, which offer powerful capabilities for real-time and continuous analysis of biofluids, mainly sweat, and can be used in athletics, consumer wellness, military, and healthcare applications. Sweat is an attractive biofluid in which different biomarkers could be noninvasively measured to provide rapid information about the physical state of an individual. Wearable devices reported so far often provide discrete (single) measurements of the target analytes, most of them in the form of a yes/no qualitative response. However, quantitative biomarker analysis over certain periods of time is highly demanded for many applications such as the practice of sports or the precise control of the patient status in hospital settings. For this, a feasible combination of fluidic elements and sensor architectures has been sought. In this regard, this paper shows a concise overview of analytical tools based on the use of capillary-driven fluidics taking place on paper or fabric devices integrated with solid-state sensors fabricated by thick film technologies. The main advantages and limitations of the current technologies are pointed out together with the progress towards the development of functional devices. Those approaches reported in the last decade are examined in detail.


Assuntos
Técnicas Biossensoriais , Dispositivos Eletrônicos Vestíveis , Biomarcadores , Humanos , Suor , Têxteis
20.
Sensors (Basel) ; 10(1): 475-90, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-22315551

RESUMO

The particular analytical performance of ultramicroelectrode arrays (UMEAs) has attracted a high interest by the research community and has led to the development of a variety of electroanalytical applications. UMEA-based approaches have demonstrated to be powerful, simple, rapid and cost-effective analytical tools for environmental analysis compared to available conventional electrodes and standardised analytical techniques. An overview of the fabrication processes of UMEAs, their characterization and applications carried out by the Spanish scientific community is presented. A brief explanation of theoretical aspects that highlight their electrochemical behavior is also given. Finally, the applications of this transducer platform in the environmental field are discussed.


Assuntos
Técnicas de Química Analítica/instrumentação , Monitoramento Ambiental/instrumentação , Análise em Microsséries/instrumentação , Microeletrodos , Transdutores , Técnicas de Química Analítica/métodos , Desenho de Equipamento , Análise de Falha de Equipamento
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