RESUMEN
This contribution describes the development of a simple, fast, cost-effective, and sensitive impedimetric immunosensor for quantifying bovine tuberculosis (TB) in bovine serum samples. The construction of the immunosensor involved immobilizing the purified protein derivative (PPD) of M. bovis onto a screen-printed electrode that was modified with gold nanoparticles (AuNPs) and a polypyrrole (pPy) film synthesized electrochemically. The immunosensor exhibited a linear range from 0.5 µg mL-1 to 100 µg mL-1 and achieved a limit of detection (LD) of 100 ng mL-1 for the detection of anti-M. bovis antibody. The recovery percentages obtained in bovine serum samples were excellent, ranging between 98 % and 103 %. This device presents several advantages over alternative methods for determining TB in bovine serum samples. These include direct, in situ measurement without the need for pre-treatment, utilization of small volumes, thus avoiding harmful solvents and expensive reagents, and portability. In addition, the immunosensor exhibits both physical and chemical stability, retaining effectiveness even after 30 days of modification. This allows simultaneous incubations and facilitates large-scale detection. Hence, this immunosensor presents itself as a promising diagnostic tool for detecting anti-M. bovis antibodies in bovine serum. It serves as a viable alternative to tuberculin and ELISA tests.
Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Oro , Nanopartículas del Metal , Tuberculosis Bovina , Animales , Bovinos , Tuberculosis Bovina/diagnóstico , Tuberculosis Bovina/sangre , Tuberculosis Bovina/inmunología , Oro/química , Técnicas Electroquímicas/métodos , Inmunoensayo/métodos , Técnicas Biosensibles/métodos , Nanopartículas del Metal/química , Mycobacterium bovis/inmunología , Polímeros/química , Pirroles/química , Electrodos , Límite de Detección , Anticuerpos Antibacterianos/sangre , Anticuerpos Antibacterianos/inmunologíaRESUMEN
Hypervalent tellurium compounds have a particular reactivity towards thiol compounds which are related to their biological properties. In this work, this property was assembled to tellurium-functionalized surfaces. These compounds were used as linkers in the immobilization process of thiolated biomolecules (such as DNA) on microcantilever surfaces. The telluride derivatives acted as reversible binding agents due to their redox properties, providing the regeneration of microcantilever surfaces and allowing their reuse for further biomolecules immobilizations, recycling the functional surface. Initially, we started from the synthesis of 4-((3-((4-methoxyphenyl) tellanyl) phenyl) amino)-4-oxobutanoic acid, a new compound, which was immobilized on a silicon surface. In nanomechanical systems, the detection involved a hybridization study of thiolated DNA sequences. Fluorescence microscopy technique was used to confirm the immobilization and removal of the telluride-DNA system and provided revealing results about the potentiality of applying redox properties to chalcogen derivatives at surfaces.
Asunto(s)
Técnicas Biosensibles , ADN/química , Silicio/química , Telurio/química , Secuencia de Bases/genética , Nanoestructuras/química , Hibridación de Ácido Nucleico , Compuestos de Sulfhidrilo/química , Propiedades de SuperficieRESUMEN
Elevated levels of copper have been detected in various types of human cancer cells, such as breast cancer cells, and a number of mechanisms have been proposed to explain the action and influence of copper on tumor progress. In this work, we found that stimulating the proliferation of mammary epithelial MCF7 cells with the high-redox-potential copper complex Cu (GlyGlyHis) is associated with the copper-induced intracellular generation of reactive oxygen species (ROS) that induces lipid peroxidation and causes increased roughness of external cell membranes, which leads to the formation of larger cell domes. The results presented herein provide new insights into the molecular link between copper and the proliferation of breast cancer cells and, consequently, into the mechanism by which changes in redox balance and ROS accumulation regulates cell membrane roughness.
Asunto(s)
Cobre/farmacología , Oligopéptidos/química , Especies Reactivas de Oxígeno/metabolismo , Western Blotting , Cobre/química , Cobre/metabolismo , Humanos , Peroxidación de Lípido/efectos de los fármacos , Células MCF-7 , Microscopía de Fuerza Atómica , Espectrofotometría AtómicaRESUMEN
The electrochromic behavior of iron complexes derived from tetra-2-pyridyl-1,4-pyrazine (TPPZ) and a hexacyanoferrate species in polyelectrolytic multilayer adsorbed films is described for the first time. This complex macromolecule was deposited onto indium-tin oxide (ITO) substrates via self-assembly, and the morphology of the modified electrodes was studied using atomic force microscopy (AFM), which indicated that the hybrid film containing the polyelectrolyte multilayer and the iron complex was highly homogeneous and was approximately 50 nm thick. The modified electrodes exhibited excellent electrochromic behavior with both intense and persistent coloration as well as a chromatic contrast of approximately 70%. In addition, this system achieved high electrochromic efficiency (over 70 cm(2) C(-1) at 630 nm) and a response time that could be measured in milliseconds. The electrode was cycled more than 10(3) times, indicating excellent stability.
RESUMEN
The present paper deals with the immobilization of redox mediators and proteins onto protected porous silicon surfaces to obtain their direct electrochemical reactions and to retain their bioactivities. This paper shows that MP-11 and viologens are able to establish chemical bonds with 3-aminopropyltriethoxylsilane-modified porous silicon surface. The functionalization of the surfaces have been fully characterized by energy dispersive X-ray analysis (EDX) and X-ray photoelectron spectroscopy (XPS) to examine the immobilization of these mediators onto the solid surface. Amperometric and open circuit potential measurements have shown the direct electron transfer between glucose oxidase and the electrode in the presence of the viologen mediator covalently linked to the 3-aminopropyltriethoxylsilane (APTES)-modified porous silicon surfaces.
Asunto(s)
Oxígeno/química , Silicio/química , Aspergillus niger/metabolismo , Electrones , Glucosa/análisis , Glucosa Oxidasa/química , Espectroscopía de Resonancia Magnética , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Modelos Químicos , Oxidación-Reducción , Propilaminas , Silanos/química , Espectrometría por Rayos X , Propiedades de Superficie , Factores de TiempoRESUMEN
The aim of this work is to study the adsorption-desorption process of bovine serum albumin (BSA) on carbon nanotubes (CNT) by reflectometry. The effect of the surface properties was analyzed by comparing the behavior of BSA on silica. The experiments were performed by reflectometry at different BSA concentrations, at pH 3.0, 4.8, and 7.0 and at two ionic strengths. Protein desorption was induced by either dilution with buffer or the addition of SDS. The initial adsorption rate is controlled by the attachment of BSA molecules to the surface, and strongly diminishes at pH 7. Adsorption isotherms reflect the high affinity of BSA for both sorbent surfaces and reach well-defined plateau values that depend on the pH, being the highest at pH 4.8 on CNT. Experiments performed at different ionic strengths (NaCl added) showed a less pronounced effect. Dilution does not induce desorption on either surface however, the addition of SDS removes protein only from the silica surface.
Asunto(s)
Nanotubos de Carbono , Albúmina Sérica Bovina/farmacocinética , Adsorción , Animales , BovinosRESUMEN
In the present work, the synthesis and the spectroelectrochemical characterization of a novel iron compound derived of tetra-2-pyridyl-1,4-pyrazine (TPPZ) with hexacyanoferrate species forming a very stable supramolecular complex in the presence of polypyrrole (PPY) matrix, is described. The hybrid material has shown excellent catalytic activity towards H(2)O(2) detection that makes it suitable for being used as redox mediator in a glucose biosensor. The hybrid FeTPPZFeCN/PPY film presents satisfactory detection limits and high sensitivity for H(2)O(2) in the presence of K(+) or Na(+) ions. For the glucose biosensor, a linear range up to 1.1 mmoll(-1) of glucose was observed with no interferences. In this case, the sensitivities obtained were 7.88 and 5.90 microAmmol(-1)lcm(-2) in phosphate buffer or NaCl solutions, respectively. The good sensitivity is related to the presence of a high-dimensional structure based on polypyridine type ligands providing an "electron antennae effect" facilitating electron tunneling between the protein and the electrode.
Asunto(s)
Técnicas Biosensibles/instrumentación , Ferrocianuros , Glucosa/análisis , Polímeros , Pirazinas , Pirroles , Compuestos Férricos/química , Peróxido de Hidrógeno/análisisRESUMEN
A copper containing Prussian Blue analogue was incorporated into a conducting polypyrrole film. The modified electrode was synthesized through an electrochemical two-step methodology leading to very stable and homogeneous hybrid films. These electrodes were proved to show excellent catalytic properties towards H(2)O(2) detection, with a performance higher than those observed for Prussian Blue and other analogues. Electrochemical impedance spectroscopy experiments demonstrated that the excellent performance of these hybrid films is strongly related to the electronic conductivity of the polymeric matrix that is wiring the copper hexacyanoferrate sites. A glucose biosensor was built-up by the immobilization of glucose oxidase; the sensitivity obtained being higher than other biosensors reported in the literature even in Na(+) containing electrolytes.
RESUMEN
Prussian blue (PB) particles with the size of ca. 5 nm were synthesized and immobilized in a multilayer structure, as a strategy for the potential development of an amperometric transducer for oxidase-enzyme-based biosensors. Multilayer films composed of PB and poly(allylamine hydrochloride) (PAH) were prepared via layer-by-layer (LbL) sequential deposition. The process was carefully monitored by UV-vis spectroscopy and cyclic voltammetry. The increase of the redox current peaks during the layer-by-layer deposition demonstrated that charge propagation within the film occurs. Linear increase of UV-vis absorbance with the number of deposited bilayers indicates that well-organized systems have been elaborated. ITO electrodes coated with PB/PAH films were used successfully for detecting H2O2, sensitivity being dependent on the number of PB/PAH layers.
Asunto(s)
Técnicas Biosensibles , Ferrocianuros/síntesis química , Nanoestructuras/química , Adsorción , Técnicas Biosensibles/instrumentación , Electroquímica , Enzimas Inmovilizadas/química , Diseño de Equipo , Ferrocianuros/química , Oxidorreductasas/químicaRESUMEN
Glucose oxidase (GOD) has been immobilized in Layer-by-Layer (LbL) films, adsorbed alternately with poly(allylamine) hydrochloride (PAH) layers, onto an ITO substrate modified with a Prussian Blue (PB) layer. The ITO/PB/GOD-PAH heterostructures were tested in amperometric glucose biosensors, with a high sensitivity of 16 microA mmol(-1)lcm(-2) and a limit of detection of 0.20 mmoll(-1) being achieved. This high sensitivity is attributed to the ultrathin nature of the film in addition to the low operating potentials that could be used due to the efficient catalysis of H(2)O(2) produced in the enzymatic reaction in the presence of Prussian Blue. The biosensors are highly selective to glucose, as demonstrated by the lack of interference from possible interferents such as ascorbic and uric acids and acetominophen. The stability of the biosensors was checked by observing an almost constant sensitivity for a period of approximately 20 days, thus indicating a stable adsorption of GOD.
Asunto(s)
Técnicas Biosensibles/instrumentación , Electroquímica/instrumentación , Análisis de Falla de Equipo/métodos , Glucosa Oxidasa/química , Glucosa/análisis , Glucosa/química , Membranas Artificiales , Adsorción , Técnicas Biosensibles/métodos , Mezclas Complejas/análisis , Electroquímica/métodos , Enzimas Inmovilizadas/química , Diseño de Equipo , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
The optimization of a biosensor prepared by the immobilization of oxalate oxidase (OOX) with a cross-linking agent onto a multilayer inorganic/organic modified electrode, is presented. A very thin Prussian Blue (PB) film covered by a self-doped polyaniline (SPAN) layer acts as very sensitive amperometric sensor for the H(2)O(2) formed by the enzymatic reaction. The electrode allows the very reliable and sensitive oxalate detection in the 0.08 to 0.45mmoll(-1) concentration range. The observed sensitivity was 131.3muAmmol(-1)cm(-2) at the operation potential of 0.05V versus Ag/AgCl in a succinate buffer solution (pH=3.8). The bilayer Prussian blue/SPAN leads to a very stable, sensitive and selective system that not only minimizes the interference caused by ascorbic and uric acids but also forms a very adherent sensing film that allows repetitive successive determinations.