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1.
Nanotechnology ; 33(27)2022 Apr 20.
Artículo en Inglés | MEDLINE | ID: mdl-35313287

RESUMEN

Herein, an amphiphilic block copolymer CD44-targeting peptide-conjugated polyethylene glycol-block-hydroxyethyl starch-block-poly (L-lactic acid) (CD44p-conjugated PEG-b-HES-b-PLA) are synthesized, which could self-assemble into the pH-responsive and CD44-targeting polymer micelles against breast cancer cells MDA-MB-231. Emodin (Emo) is a natural anthraquino with pharmacological activities in anti-tumor effects. However, Emo suffers from poor water solubility, low biocompatibility, rapid systemic elimination, and off-target side effects, resulting in unsatisfactory treatment outcomes. Nanotechnology-based drug delivery systems have proven great potential for cancer chemotherapy. The constructed polymeric micelles Emo@CD44p-PM have exhibited an average size of 154.5 ± 0.9 nm characterized by DLS and TEM. Further, the Emo@CD44p-PM have effective Emo-loading capacity, good thermal stability, and pH responsiveness. Intracellular uptake study shows the enhanced cellular internalization of Emo@CD44p-PM due to the increased exposure of CD44p enhances the cellular internalization of Emo@CD44p-PM effectively. Furthermore, thein vitroresults showed Emo@CD44p-PM has been observed good biocompatibility and anti-tumor effects. Therefore, the polymeric micelles Emo@CD44p-PM provide a promising delivery strategy of targeted therapy for breast cancer.


Asunto(s)
Neoplasias de la Mama , Emodina , Neoplasias de la Mama/tratamiento farmacológico , Línea Celular Tumoral , Portadores de Fármacos , Sistemas de Liberación de Medicamentos , Femenino , Humanos , Receptores de Hialuranos , Concentración de Iones de Hidrógeno , Micelas , Poliésteres , Polietilenglicoles , Polímeros
2.
Anal Chem ; 93(31): 11010-11018, 2021 08 10.
Artículo en Inglés | MEDLINE | ID: mdl-34323073

RESUMEN

Conventional photosensitive materials such as TiO2 suffer from restricted absorption in the ultraviolet region, fast recombination of photogenerated electron-hole pairs, and a lack of functional groups for biocoupling, which hinder their application in photoelectrochemical (PEC) biosensing. Herein, a new coordination polymer (CP) based on Cu(I), chloridion, and 4-mercaptobenzoic acid (MBA) has been designed and synthesized (called CuClx(MBA)y). The prepared p-type CuClx(MBA)y exhibits visible-light absorption due to its narrow optical band gap (2.59 eV), and its proper band edge position enables it to form a p-n junction with TiO2. Through layer-by-layer assembling, the photocurrent intensity of the CuClx(MBA)y/TiO2/FTO composite photoelectrode was 3.7-fold higher than that of a TiO2/FTO electrode and 35-fold higher than a CuClx(MBA)y/FTO electrode. The potential enhancement mechanism was discussed, which lies in the contributions of CuClx(MBA)y in enhancing absorption in the visible-light region and boosting the separation of electron-hole pairs of TiO2 by the p-n junction. Furthermore, CuClx(MBA)y nanosheets can realize bioconjugation directly, thanks to its abundant carboxyl groups. The CuClx(MBA)y/TiO2/FTO composite photoelectrodes were applied to develop a sensitive PEC biosensor for microRNA-21 (model target). By subtly exploiting the energy transfer between CuClx(MBA)y and Au nanoparticles (AuNPs), AuNPs served as effective quenchers. In the presence of the target, AuNP-labeled sDNA1 connected to the electrode surface, and thus, a decreased photocurrent was obtained. The proposed biosensor has a low detection limit of 0.29 fM (S/N = 3), good selectivity, and reproducibility. The proposed system was applied to monitor microRNA in cancer cells with satisfying results.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , MicroARNs , Técnicas Electroquímicas , Oro , Límite de Detección , Polímeros , Reproducibilidad de los Resultados , Titanio
3.
Anal Chem ; 93(49): 16727-16733, 2021 12 14.
Artículo en Inglés | MEDLINE | ID: mdl-34851090

RESUMEN

Surface-enhanced Raman scattering (SERS) is a super-sensitive analysis technology based on the target molecular fingerprint information. The enhancement of local electromagnetic field of the SERS substrate would increase the target molecules' Raman intensity which adsorb on the surface of nanoparticles. However, the existing adhesive macromolecules in the complex mixed sample would interfere with the adsorption of small target molecules, and it weakens the Raman intensity of target molecules. Microgels are one of the potential materials to suppress the interference of adhesive macromolecules and to avoid the complex pretreatments. However, most of the current microgel synthesis methods involve complex operations with precise instrumentation or the interference of oil and organic reagents. In this work, a simple and oil-free method was proposed to synthesize the gold nanobipyramid (Au NBP)@Ag@hyaluronic acid microgel via the condensation reaction of carboxyl and amino groups. As a proof-of-concept demonstration for small-molecule detection, the rhodamine 6G (R6G) molecules were allowed to enter inside the microgel through the meshes and adsorb on the surface of Au NBP@Ag nanoparticles within 30 min, while the macromolecule (bovine serum albumin in this case) was retained outside the microgel in the meantime. In addition, under the combined action of lightning rod effect of Au NBP and surface plasmon resonance effect of silver render the microgels with high SERS activity. The synthetic Au NBP@Ag@hyaluronic acid microgels were applied to detect 6-thioguanine in the human serum without any pretreatment process, and it showed a high signal enhancement and stable SERS signal, which can satisfy the requirement of clinical diagnosis. These results show that the proposed microgels have potential applications in the field of point-of-care testing.


Asunto(s)
Nanopartículas del Metal , Microgeles , Oro , Humanos , Plata
4.
Anal Chem ; 91(18): 11821-11826, 2019 09 17.
Artículo en Inglés | MEDLINE | ID: mdl-31436088

RESUMEN

The development of simple but sensitive methods for hyaluronidase (HAase) detection has been paid a great deal of attention because HAase is a potential cancer marker. In this work, a novel system coupled with a controlled release system has been designed for HAase determination without complex analytical instruments and skilled technicians. Pt@SiO2 nanoparticles (NPs), which can catalyze the breakdown of H2O2 into O2 and H2O, was embedded in the hydrogel constructed by polyethylenimine (PEI) and hyaluronic acid (HA). In the presence of HAase, the hydrogel was broken down as HAase can catalyze the degradation of HA and hence the Pt@SiO2 NPs in the hydrogel was released. The released Pt@SiO2 NPs mixed with H2O2 solution in a drainage device, and then O2 was generated due to the decomposition of H2O2, resulting in an enhancement of pressure in the drainage device because of the low solubility of O2. A certain amount of H2O overflowed from the drainage device because the difference of the pressure between the inner and outer of the drainage device. The overflowed H2O was collected by a tube, and its amount was easily measured by an electronic balance. The weight of the H2O has a linear relationship with the HAase concentration in the range of 1-60 U/mL (120 min enzymatic hydrolysis time) and 0.2-10 U/mL (240 min enzymatic hydrolysis time). The developed system has been applied to detect the activity of HAase in urine samples with satisfied results.


Asunto(s)
Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/métodos , Hialuronoglucosaminidasa/análisis , Hidrogeles/química , Electrones , Diseño de Equipo , Humanos , Ácido Hialurónico/química , Hialuronoglucosaminidasa/orina , Hidrogeles/síntesis química , Peróxido de Hidrógeno/química , Concentración de Iones de Hidrógeno , Nanopartículas del Metal/química , Microscopía Electrónica de Transmisión , Neoplasias/enzimología , Neoplasias/orina , Oxígeno/química , Polietileneimina/química , Temperatura
5.
Anal Chem ; 88(4): 2345-52, 2016 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-26765320

RESUMEN

A disposable, equipment-free, versatile point-of-care testing platform, microfluidic distance readout sweet hydrogel integrated paper-based analytical device (µDiSH-PAD), was developed for portable quantitative detection of different types of targets. The platform relies on a target-responsive aptamer cross-linked hydrogel for target recognition, cascade enzymatic reactions for signal amplification, and microfluidic paper-based analytic devices (µPADs) for visual distance-based quantitative readout. A "sweet" hydrogel with trapped glucoamylase (GA) was synthesized using an aptamer as a cross-linker. When target is present in the sample, the "sweet" hydrogel collapses and releases enzyme GA into the sample, generating glucose by amylolysis. A hydrophilic channel on the µPADs is modified with glucose oxidase (GOx) and colorless 3,3'-diaminobenzidine (DAB) as the substrate. When glucose travels along the channel by capillary action, it is converted to H2O2 by GOx. In addition, DAB is converted into brown insoluble poly-3,3'-diaminobenzidine [poly(DAB)] by horseradish peroxidase, producing a visible brown bar, whose length is positively correlated to the concentration of targets. The distance-based visual quantitative platform can detect cocaine in urine with high selectivity, sensitivity, and accuracy. Because the target-induced cascade reaction is triggered by aptamer/target recognition, this method is widely suitable for different kinds of targets. With the advantages of low cost, ease of operation, general applicability, and disposability with quantitative readout, the µDiSH-PAD holds great potential for portable detection of trace targets in environmental monitoring, security inspection, personalized healthcare, and clinical diagnostics.


Asunto(s)
Hidrogel de Polietilenoglicol-Dimetacrilato/química , Técnicas Analíticas Microfluídicas , Papel , Pruebas en el Punto de Atención , Cocaína/orina , Reactivos de Enlaces Cruzados/química , Reactivos de Enlaces Cruzados/metabolismo , Equipos Desechables , Glucano 1,4-alfa-Glucosidasa/metabolismo , Glucosa/metabolismo , Glucosa Oxidasa/metabolismo , Peroxidasa de Rábano Silvestre/metabolismo , Hidrogel de Polietilenoglicol-Dimetacrilato/metabolismo
6.
Anal Chem ; 87(8): 4275-82, 2015 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-25806667

RESUMEN

A versatile point-of-care assay platform was developed for simultaneous detection of multiple targets based on a microfluidic paper-based analytic device (µPAD) using a target-responsive hydrogel to mediate fluidic flow and signal readout. An aptamer-cross-linked hydrogel was used as a target-responsive flow regulator in the µPAD. In the absence of a target, the hydrogel is formed in the flow channel, stopping the flow in the µPAD and preventing the colored indicator from traveling to the final observation spot, thus yielding a "signal off" readout. In contrast, in the presence of a target, no hydrogel is formed because of the preferential interaction of target and aptamer. This allows free fluidic flow in the µPAD, carrying the indicator to the observation spot and producing a "signal on" readout. The device is inexpensive to fabricate, easy to use, and disposable after detection. Testing results can be obtained within 6 min by the naked eye via a simple loading operation without the need for any auxiliary equipment. Multiple targets, including cocaine, adenosine, and Pb(2+), can be detected simultaneously, even in complex biological matrices such as urine. The reported method offers simple, low cost, rapid, user-friendly, point-of-care testing, which will be useful in many applications.


Asunto(s)
Adenosina/análisis , Cocaína/análisis , ADN/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Plomo/análisis , Técnicas Analíticas Microfluídicas , Papel , ADN/síntesis química , ADN/aislamiento & purificación , Hidrogel de Polietilenoglicol-Dimetacrilato/síntesis química , Hidrogel de Polietilenoglicol-Dimetacrilato/aislamiento & purificación , Estructura Molecular , Compuestos Organofosforados/síntesis química , Compuestos Organofosforados/química , Sistemas de Atención de Punto
7.
Anal Chem ; 87(18): 9204-8, 2015 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-26295334

RESUMEN

Traditional electrochemical DNA biosensors need DNA immobilization on the electrode surface, which is tedious and time-consuming. In this study, a simple but ultraselective electrochemical DNA biosensor had been designed to determine target DNA species related to oral cancer overexpressed 1 in saliva, which combines the signal amplification of nicking endonuclease assisted target recycling with the immobilization-free electrochemical method. The complementary substrate strand of target DNA species contains a simple asymmetric sequence had been modified with a methylene blue at the 3' terminal first, which cannot diffuse easily to the negative charged ITO electrode surface due to the abundant negative charges. The presence of the target DNA would trigger the formation of double-stranded DNA (dsDNA). Then the nicking endonuclease can recognize the simple asymmetric sequence in the dsDNA and cleave the substrate strand of ds-DNA into two pieces, a long ssDNA and a 2-base ssDNA linked with methylene blue. The short one can diffuse easily to the negative charged ITO electrode surface and results in the enhanced electrochemical response detected. At the same time, the target DNA can dissociate from the dsDNA and trigger the next round of hybridization, cleavage, and releasing, which results in the signal amplification. This homogeneous DNA biosensor can detect as low as 0.35 pM (S/N = 3) target DNA. Compared with the traditional heterogeneous electrochemical DNA biosensors, which are tedious and time-consuming due to the complex DNA immobilization process, the assay not only owns the merits of simple and high efficiency since performed in a homogeneous solution but also exhibits a high distinction ability to single-base mismatch, double-bases mismatch, and noncomplementary DNA sequence.


Asunto(s)
Técnicas Biosensibles/métodos , ADN/análisis , ADN/genética , Desoxirribonucleasa I/metabolismo , Neoplasias de la Boca/genética , Técnicas de Amplificación de Ácido Nucleico , Disparidad de Par Base , Secuencia de Bases , Materiales Biomiméticos/química , Calibración , Electroquímica , Límite de Detección , Polimorfismo de Nucleótido Simple , Saliva/química , Compuestos de Estaño/química
8.
Talanta ; 276: 126203, 2024 Aug 15.
Artículo en Inglés | MEDLINE | ID: mdl-38718648

RESUMEN

T-2 toxin, a hazardous mycotoxin often present in cereals and products based on cereals, poses a substantial risk to humans and animals due to its high toxicity. The development of uncomplicated, quick and highly sensitive methods for detecting T-2 toxin is imperative. In this work, a portable sensing system was constructed using water column height as a readout device in combination with a controlled release system, which allows for an accurate quantitative analysis of T-2 toxin without the need for expensive instrumentation or skilled technicians. Hyaluronic acid (HA) hydrogel was constructed by double cross-linked DNA/aptamer hybrids with polyethyleneimine (PEI) and embedded with platinum nanoparticles (Pt NPs). The aptamer specifically bound to T-2 toxin in its presence, resulting in the disruption of the hydrogel and subsequent release of the Pt NPs. These Pt NPs were later mixed with a solution of H2O2 in a confined reaction flask, leading to the decomposition of H2O2 into O2. A glass capillary tube containing a column of red water had been inserted into the cap of the reaction flask, and the low solubility of O2 led to an increase in pressure within the reaction unit, causing the red water column to rise. There is a good linear correlation between the height of the capillary liquid level and the T-2 toxin concentration in the range of 20 ng/mL to 6 µg/mL. The system has been successfully used to detect T-2 toxin in samples of barley tea and corn.


Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Nanopartículas del Metal , Platino (Metal) , Toxina T-2 , Toxina T-2/análisis , Técnicas Biosensibles/métodos , Aptámeros de Nucleótidos/química , Nanopartículas del Metal/química , Platino (Metal)/química , Agua/química , ADN/química , ADN/análisis , Hidrogeles/química , Límite de Detección , Ácido Hialurónico/química , Polietileneimina/química
9.
Adv Healthc Mater ; 13(9): e2303379, 2024 04.
Artículo en Inglés | MEDLINE | ID: mdl-38211342

RESUMEN

Hydrogel dressings capable of infection monitoring and precise treatment administration show promise for advanced wound care. Existing methods involve embedd ingorganic dyes or flexible electronics into preformed hydrogels, which raise safety issues and adaptability challenges. In this study, an injectable hydrogel based smart wound dressing is developed by integrating food-derived anthocyanidin as a visual pH probe for infection monitoring and poly(L-lactic acid) microcapsules as ultrasound-responsive delivery systems for antibiotics into a poly(ethylene glycol) hydrogel. This straightforwardly prepared hydrogel dressing maintains its favorable properties for wound repair, including porous morphology and excellent biocompatibility. In vitro experiments demonstrated that the hydrogel enabled visual assessment of pH within the range of 5 âˆ¼ 9.Meanwhile, the release of antibiotics could be triggered and controlled by ultrasound. In vivo evaluations using infected wounds and diabetic wounds revealed that the wound dressing effectively detected wound infection by monitoring pH levels and achieved antibacterial effects through ultrasound-triggered drug release. This led to significantly enhanced wound healing, as validated by histological analysis and the measurement of inflammatory cytokine levels. This injectable hydrogel-based smart wound dressing holds great potential for use in clinical settings to inform timely and precise clinical intervention and in community to improve wound care management.


Asunto(s)
Vendajes , Hidrogeles , Hidrogeles/química , Cápsulas , Antibacterianos/farmacología , Antibacterianos/química , Materiales Biocompatibles , Concentración de Iones de Hidrógeno
10.
J Sep Sci ; 36(3): 477-84, 2013 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-23292891

RESUMEN

An RP LC-ESI-MS/MS method for the determination of the migration of 16 primary phthalic acid esters from plastic samples has been developed using distilled water, 3% acetic acid, 10% alcohol, and olive oil as food simulants. Detection limits were 1.6-18.5 µg/kg in distilled water, 1.4-17.3 µg/kg in 3% acetic acid, 1.4-19.2 µg/kg in 10% alcohol, and 31.9-390.8 µg/kg in olive oil. The RSDs were in the range of 0.07-11.28%. The real plastic products inspection showed that only few analyzed samples were phthalates contaminated. Bis-2-ethylhexyl ester and dibutyl phthalate were the common items migrated from the plastic products into food and feeds, but the migration concentrations were far below the limits set by European Union (1.5 mg/kg for bis-2-ethylhexyl ester and 0.3 mg/kg for dibutyl phthalate).


Asunto(s)
Cromatografía Liquida/métodos , Ésteres/química , Contaminación de Alimentos/análisis , Embalaje de Alimentos/instrumentación , Ácidos Ftálicos/química , Aceites de Plantas/análisis , Plásticos/análisis , Espectrometría de Masas en Tándem/métodos , Aceite de Oliva
11.
ACS Sens ; 8(2): 884-892, 2023 02 24.
Artículo en Inglés | MEDLINE | ID: mdl-36657970

RESUMEN

The development of an instrument-free, on-site, real-time, sensitive, and visualized fluoride-ion (F-) content rapid detection strategy is crucial to ensuring the health of the population. Smart microdevices that are portable, directly read, and easy to operate have recently attracted much attention. Herein, a ratiometric fluorescent probe (AA-CDs@[Ru(bpy)3]2+)-based smartphone sensing platform was developed for the detection of F-. The red fluorescent ruthenium bipyridine [Ru(bpy)3]2+ molecule was chosen as the reference signal, and the carbon dots (AA-CDs) with Al3+ aggregation-induced enhanced emission (AIE) were designed as the response signal. The ratiometric probe fluorescence changed continuously from red to cyan in response to different concentrations of F-, and the red-green-blue (RGB) channel values of the fluorescence image were extracted through the smartphone color recognition application (APP). There was a linear relationship between the blue-red (B/R) ratio and the F- concentration, with a limit of detection (LOD) of 1.53 µM, far below the allowable content of F- in drinking water prescribed by the World Health Organization. The F- content was rapidly detected on-site with satisfactory repeatability and relative standard deviation using several water and toothpaste samples as the real sample. The platform features low cost, portability, easy operation, and good stability, selectivity, and repeatability, which provides a powerful tool for the visual quantitative detection of smartphone-based microsensing platforms possibly in the fields of environmental protection, diagnosis, and food safety assessment.


Asunto(s)
Fluoruros , Teléfono Inteligente , Carbono , Inocuidad de los Alimentos
12.
Luminescence ; 26(6): 531-5, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-22162456

RESUMEN

The electrochemiluminescence (ECL) behavior of luminol on a nafion-nano-TiO(2) modified glassy carbon electrode (nafion-nano-TiO(2)--GCE) was studied. Two ECL peaks (ECL-1 and ECL-2) were found during cathodic potential scanning. ECL-1 at ca -0.4 V (vs Ag--AgCl reference electrode) came from the reaction between luminol and active oxygen anion produced at the GCE surface directly, while ECL-2 at ca -0.9 V (vs Ag--AgCl reference electrode) came from the reaction between luminol and the active oxygen anion catalyzed by TiO(2.) The possible mechanism for the generation of both ECL peaks has been proposed. The reproducibility of the ECL intensities on nafion-nano-TiO(2)--GCE at ECL-1 and ECL-2 was good, with relative standard deviations (n = 10) of 4.3 and 1.3%, respectively. The ECL-2 generated at the nafion-nano-TiO(2)--GCE surface was further developed to detect the dissolved oxygen, and a detection limit of 0.02 mg/L was achieved. The proposed method was applied to detect dissolved oxygen in water with satisfactory result.


Asunto(s)
Electroquímica , Electrodos , Polímeros de Fluorocarbono/química , Luminol/química , Nanopartículas del Metal , Titanio/química , Concentración de Iones de Hidrógeno , Luminiscencia , Reproducibilidad de los Resultados
13.
Int J Nanomedicine ; 15: 9061-9074, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33239874

RESUMEN

PURPOSE: Therapy for triple-negative breast cancer (TNBC) is a global problem due to lack of specific targets for treatment selection. Cancer stem cells (CSCs) are responsible for tumor formation and recurrence but also offer a promising target for TNBC-targeted therapy. Here, zirconium-89 (89Zr)-labelled multifunctional liposomes (MLPs) surface-decorated with chitosan (CS) were fabricated to specifically target and trace cluster of differentiation 44+ (CD44+) TNBC CSCs specifically. PATIENTS AND METHODS: The biological basis of CS targeting CD44 for cancer therapy was investigated by detecting the expression of CD44 in TNBC CSCs and TNBC tissues. Molecular docking and dynamics simulations were performed to investigate the molecular basis of CS targeting CD44 for cancer therapy. Gambogic acid (GA)-loaded, 89Zr@CS-MLPs (89Zr-CS-GA-MLPs) were prepared, and their uptake and biodistribution were observed. The anti-tumor efficacy of 89Zr@CS-GA-MLPs was investigated in vivo. RESULTS: CD44 is overexpressed in TNBC CSCs and tissues. Molecular docking and dynamics simulations showed that CS could be stably docked into the active site of CD44 in a reasonable conformation. Furthermore, 89Zr@CS-GA-MLPs were able to bind specifically to CD44+ TNBC stem-like cells and accumulated in tumors of xenograft-bearing mice with excellent radiochemical stability. 89Zr@CS-GA-MLPs loaded with GA showed remarkable anti-tumor efficacy in vivo. CONCLUSION: The GA-loaded, 89Zr-labelled, CS-decorated MLPs developed in this study represent a novel strategy for TNBC imaging and therapy.


Asunto(s)
Liposomas/química , Células Madre Neoplásicas/efectos de los fármacos , Radioisótopos/química , Neoplasias de la Mama Triple Negativas/diagnóstico por imagen , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Circonio/química , Adulto , Anciano , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Línea Celular Tumoral , Quitosano/química , Femenino , Humanos , Receptores de Hialuranos/química , Receptores de Hialuranos/metabolismo , Liposomas/farmacocinética , Ratones Desnudos , Persona de Mediana Edad , Simulación del Acoplamiento Molecular , Terapia Molecular Dirigida/métodos , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Tomografía de Emisión de Positrones/métodos , Distribución Tisular , Neoplasias de la Mama Triple Negativas/patología , Xantonas/farmacocinética , Ensayos Antitumor por Modelo de Xenoinjerto
14.
Angew Chem Int Ed Engl ; 48(11): 2044-6, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19191275

RESUMEN

To each his own: An addressable electrochemical device consisting of orthogonally arranged rows and columns of electrodes has been constructed to monitor protein expression in genetically engineered cells at the single-cell level. The response based on redox cycling reflected the different expression levels of the enzyme from individual HeLa cells transfected with a plasmid vector including secreted alkaline phosphatase.


Asunto(s)
Fosfatasa Alcalina/metabolismo , Técnicas Electroquímicas/métodos , Fosfatasa Alcalina/genética , Aminofenoles/química , Línea Celular , Técnicas Electroquímicas/instrumentación , Expresión Génica , Células HeLa , Humanos , Análisis por Micromatrices , Microelectrodos , Oxidación-Reducción , Poliestirenos/química
15.
Analyst ; 133(6): 797-801, 2008 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-18493682

RESUMEN

Nickel(ii) tetrasulfophthalo-cyanine (NiTSPc) could be electrodeposited onto an indium tin oxide (ITO) electrode to form an electropolymerization film of NiTSPc. The electrochemiluminescent (ECL) behavior of bis-[3,4,6-trichloro-2-(pentyloxy-carbonyl)-phenyl] oxalate (BTPPO) on this modified electrode was investigated in detail. The emission of ECL of BTPPO can be greatly enhanced by hydrogen peroxide at this modified electrode. Thus, a new ECL biosensor for uric acid was developed based on the enzymatic reaction of uric acid in the presence of uricase to produce H(2)O(2). The developed sensor has been used to detect uric acid in real serum samples, and the results compared well with those obtained by the routine method.


Asunto(s)
Técnicas Biosensibles , Ácido Úrico/sangre , Electroquímica/métodos , Electrodos , Humanos , Peróxido de Hidrógeno , Indoles , Isoindoles , Mediciones Luminiscentes , Níquel , Polímeros , Compuestos de Estaño
16.
Anal Chim Acta ; 1002: 90-96, 2018 Mar 09.
Artículo en Inglés | MEDLINE | ID: mdl-29306417

RESUMEN

A simple, low-cost, and sensitive liposome-based colorimetric aptasensor has been developed to detect ochratoxin A (OTA). Specifically, a dumbbell-shaped probe was designed, including magnetic beads (MBs), double-stranded DNA (dsDNA), and enzyme-encapsulated liposome. The dsDNA formed by the hybridization between OTA aptamer and its complementary probe. And the dsDNA was used to contact the MBs and the enzyme-encapsulated liposome. In the presence of OTA, the aptamer preferred to combine with OTA to form G-quadruplex, resulting in the release of the detection probe and the enzyme-encapsulated liposome. Each liposome contained a large amount of HRP. Thus, when the liposome was lysed by adding the mixed solution of 3,3',5,5'-tetramethylbenzidine (TMB) and H2O2, a large number of HRP were released. HRP could catalyze the H2O2-mediated oxidation of TMB and hence resulted in the color change from colorless to blue with the OTA concentration varying, and this variation can be observed by naked eyes easily. The result showed that the absorption intensity at 652 nm enhanced with the increase of OTA concentration ranging from 0.05 to 2.0 ng mL-1, and the limit of detection was calculated to be 0.023 ng mL-1 (S/N = 3). The developed colorimetric aptasensor has been applied to detect OTA concentration in corn samples with satisfied results.


Asunto(s)
Aptámeros de Nucleótidos/metabolismo , Técnicas Biosensibles/métodos , Colorimetría/métodos , Peroxidasa de Rábano Silvestre/metabolismo , Límite de Detección , Liposomas/química , Ocratoxinas/análisis , Cápsulas , Análisis de los Alimentos , Peroxidasa de Rábano Silvestre/química , Ocratoxinas/metabolismo , Zea mays/química
17.
J Chromatogr A ; 1172(1): 84-91, 2007 Nov 16.
Artículo en Inglés | MEDLINE | ID: mdl-17936770

RESUMEN

A new capillary electrophoresis-electrochemiluminescence (ECL) detection system equipped with an electrically heated Ru(bpy)(3)(2+)/multi-wall-carbon-nanotube paste electrode (Ru(bpy)(3)(2+)/MWNTPE) was developed. Ru(bpy)(3)(2+) was immobilized in the electrode by directly mixing with the multi-wall-carbon-nanotube paste (MWNTP). This modified electrode could be electrically heated and temperature of the electrode (Te) could be accurately controlled. Tri-n-propylamine (TPrA) was used as coreactant to investigate CE-ECL signals under different conditions. Compared with the conventional electrode at room temperature, the heated electrode has been shown to provide some advantages, such as higher sensitivity, lower RSD, and decreasing width of the peak. Furthermore, wider range of capillary-to-electrode distance and larger-area electrode are a benefit to CE-ECL. In addition, this system has been applied to separation and detection of acephate and dimethoate. The results indicated that the present CE-ECL system coupled with heated modified-electrode could provide high sensitivity, wide linear range, satisfying linear relationship and excellent reproducibility.


Asunto(s)
Electroforesis Capilar/instrumentación , Mediciones Luminiscentes/métodos , Microelectrodos , Nanotubos de Carbono , Compuestos de Rutenio/química , 2,2'-Dipiridil/análogos & derivados , Electroforesis Capilar/métodos , Electroforesis por Microchip , Estudios de Factibilidad , Polímeros de Fluorocarbono , Concentración de Iones de Hidrógeno , Compuestos Organometálicos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Temperatura
18.
Anal Chim Acta ; 963: 112-118, 2017 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-28335964

RESUMEN

Development of simple but sensitive biosensor for influenza detection is highly important in immediate and effective clinical treatment. In this study, a sensitive colorimetric immunosensor which combines the advantages of high selectivity of immunoassay and simplicity of colorimetric detection has been developed to detect influenza virus H5N1 based on enzyme-encapsulated liposome. Biotin-tagged liposome encapsulated with large amount of horseradish peroxidase (HRP) was firstly synthesized. In the presence of H5N1, H5N1 co-bound with the capture antibody and the biotinylated detection antibody to form sandwich immunocomplex. Subsequently, the HRP-encapsulated liposome was introduced to conjugate with the detection antibody through biotin-avidin-biotin linkage. Upon the addition of substrate (mixture of 3,3',5,5'-tetramethylbenzidine (TMB) and H2O2), the liposome was directly lysed to release large amount of HRP by TMB. The released HRP catalyzed the H2O2-mediated oxidation of TMB, resulting in color change of the system, which was observed by naked eyes or UV-vis spectra. The result showed that the absorption intensity enhanced with the increase of H5N1 concentration ranging from 0.1 to 4.0 ng/mL, and the detection limit was calculated to be 0.04 ng/mL. The sensitivity of the proposed biosensor is much higher than that of conventional enzyme-linked immunosorbent assay method. The proposed immunosensor is relatively simple, low-cost, sensitive, and selective without using any sophisticated instruments, therefore it may have a promising prospect for detecting targets in clinical medicine, food safety analysis, and environmental monitoring.


Asunto(s)
Técnicas Biosensibles/métodos , Colorimetría/métodos , Peroxidasa de Rábano Silvestre/química , Inmunoensayo/métodos , Subtipo H5N1 del Virus de la Influenza A/aislamiento & purificación , Límite de Detección , Liposomas/química , Cápsulas , Peroxidasa de Rábano Silvestre/metabolismo , Humanos
19.
Biosens Bioelectron ; 55: 412-6, 2014 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-24434497

RESUMEN

A novel portable biosensor for sensitive and selective detection of platelet-derived growth factor-BB (PDGF-BB) had been developed based on the specific recognition between aptamer and protein using a personal glucose meter (PGM) as readout. In the presence of PDGF-BB, the primary aptamer of PDGF-BB which is bound to the surface of streptavidin magnespheres paramagnetic particles (SA-PMPs) reacts quantitatively with invertase-functionalized secondary aptamer of PDGF-BB to form a stable complex, resulting in the attachment of invertase on the SA-PMPs. Subsequently, the invertase catalyzes the hydrolysis of sucrose to produce a large amount of glucose and quantitative readout by the PGM. The enhanced signal of the PGM has a relationship with the concentration of PDGF-BB in the range of 1.0 × 10(-14) M~3.16 × 10(-12) M, and the detection limit is 2.9 fM. The proposed portable biosensor had been successfully applied to assay the PDGF-BB in saliva samples.


Asunto(s)
Aptámeros de Nucleótidos/genética , Técnicas Biosensibles/instrumentación , Automonitorización de la Glucosa Sanguínea/instrumentación , Sistemas de Atención de Punto , Proteínas Proto-Oncogénicas c-sis/análisis , Saliva/química , Secuencia de Bases , Becaplermina , Diseño de Equipo , Análisis de Falla de Equipo , Humanos , Miniaturización , Datos de Secuencia Molecular , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
20.
Biosens Bioelectron ; 42: 612-7, 2013 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-23261698

RESUMEN

In this paper, a novel strategy of protein molecularly-imprinted electrochemical sensor was proposed to determine Bovine Hemoglobin selectively. Gold microdendrites and prussian blue particles were electrodeposited on the surface of glassy carbon electrode respectively to improve the sensitivity of MIECS. The molecularly imprinted polymers film is synthesized by electrochemically induced redox polymerization of acrylamide in the presence of template protein. The surface feature of the modified electrode was characterized by cyclic voltammetry. The reduced electrochemical intensity which was produced by prussian blue is related to the concentration of Bovine Hemoglobin. The times of washing and incubation of modified electrode were investigated and optimized. The MIECS with improved conductivity and electrochemical performances showed good response in the range of low concentrations from 0.1 to 1.0×10³ µg/mL and the detection limit (LOD, 3σ, RSD≤4.31 %) is 0.05 µg/mL. The result demonstrates significant specific recognition toward the template protein via selective test and a reasonable regeneration in five recyclings. The proposed sensor exhibited high sensitivity and selectivity, acceptable reproducibility, and could be extended to recognize other protein.


Asunto(s)
Técnicas Biosensibles/métodos , Ferrocianuros/química , Hemoglobinas/aislamiento & purificación , Animales , Bovinos , Técnicas Electroquímicas , Oro/química , Límite de Detección , Microelectrodos , Polímeros/química , Sensibilidad y Especificidad
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