RESUMEN
Photoelectrochemical sensors have outstanding advantages including high sensitivity and miniaturization for outdoor use. Recently, perovskite quantum dots have attracted significant attention due to their high photoluminescence quantum yield. Nonetheless, there is still a strong need to improve their performance in challenging aqueous biological applications. In this paper, based on the molecularly imprinted polymer encapsulation of CsPbBr3 perovskite quantum dot/TiO2 inverse opal heterojunction structures, linear photoelectrochemical detection of cholesterol in aqueous solution was obtained without the involvement of an enzyme. The attenuation of photocurrent intensity under intermittent irradiation within 900 s (45 on/off cycles) was only 8.6%, demonstrating the superior stability of CsPbBr3 based sensor here. At the same time, the minimum detection limit of 1.22 × 10-9 mol L-1 in buffer conditions was lower than that reported for cholesterol photoelectric sensors. It has also been shown that the photoelectrochemical sensor of CsPbBr3 here outperformed that of CH3NH3PbBr3, which is another important member of the perovskite family. Finally, the proposed photoelectrochemical sensor platform was successfully applied in the determination of cholesterol in challenging serum with satisfactory recovery. The synergism among CsPbBr3 perovskite quantum dots, TiO2 inverse opal structure and imprinted polymer has led to greatly improved water stability, super selectivity and sensitivity, thus promoting the development of perovskite-based biological sensors.
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Aptamers have gained significant attention as the molecular recognition element to replace antibodies in sensor development and target delivery. Nevertheless, it is noteworthy that unlike the wide application of polyvalent antibodies, existing researches on the combined use of heterologous aptamers with similar recognition affinity and specificity for target detection were sporadic. Herein, first, the wide existence of polyaptamer for bacteria was revealed through the summary of existing literature. Furthermore, based on the establishment of a sensitive aptamer cocktail/graphene oxide fluorescence resonance energy transfer polyaptasensor with a detection limit as low as 10 CFU/ml, the systemic characterization of aptamer cocktails in bacterial detection was carried out by taking E. coli, Vi. parahemolyticus, S. typhimurium, and C. sakazakii as the assay targets. It was turned out that the polyaptasensors for C. sakazakii and S. typhimurium owned prevalence in the broader concentration range of target bacteria. While the polyaptasensors for E. coli and V. parahemolyticus outperformed monoaptasensor mainly in the lower concentration of target bacteria. The linear relationships between fluorescence recovery and the concentration of bacteria were also discussed. The different characteristics of the bacterial cellular membrane, including the binding affinity and the robustness to variation, are analyzed to be the main reason for the diverse detection performance of aptasensors. The study here enhances a sensor detection strategy with super sensitivity. More importantly, this systemic study on the aptamer cocktail in reference to antibodies will advance the in-depth understanding and rational design of aptamer based biological recognition, detection, and targeting.
Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Grafito , Aptámeros de Nucleótidos/química , Escherichia coli/genética , Escherichia coli/metabolismo , Transferencia Resonante de Energía de Fluorescencia , Grafito/química , Límite de DetecciónRESUMEN
Rapid screening of active compounds plays a crucial role in the research and application of complex natural medicines. Herein, a new method of simultaneous label-free multi-drug screening based on a selective aptamer-carboxyfluorescein/graphene oxide energy transfer optical sensor combined with microfluidic chip electrophoretic separation is reported. In this study, seven traditional Chinese medicinal monomers were chosen as targets for the screening of G-quadruplex ligands. The screening results of the G-quadruplex active ligands, including daidzein, berberine hydrochloride, jatrorrhizine hydrochloride, and fangchinoline, and non-active ligands, including geniposide and oxymatrine, were consistent with those reported in literature. Moreover, one new potential G4DNA active drug, jujuboside A, was identified. Molecular simulation of the interaction between G4DNA and drugs was also carried out using HyperChem and AutoDock to verify the results of the experimental screening. It further demonstrated the reliability of our strategy. This novel separation and concentration based multi-sensing strategy provides a simple, rapid, and sensitive tool for simultaneous multi-drug screening, which is very meaningful for drug screening and bio-interaction analysis.
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Técnicas Biosensibles , Evaluación Preclínica de Medicamentos/métodos , Electroforesis , G-Cuádruplex , Microfluídica , Aptámeros de Nucleótidos , Transferencia de Energía , Fluoresceínas , Grafito , Ligandos , Reproducibilidad de los Resultados , SaponinasRESUMEN
The selective and sensitive detection of cancerous exosomes in serum is critical for early disease diagnosis and improved prognosis. Previous exosome-related research has been limited by a lack of well-understanding in exosomes as well as the challenging background interference of body fluid. Molecularly imprinted polymers (MIPs) and nucleic acid aptamers can be regarded as the two alternatives to antibodies. When using imprinted polymer technology, comprehensive and precise information about the target constituents is not required. In this study, a novel kind of dual selective fluorescent nanosensor for the poorly characterized exosomes was constructed by integrating magnetic MIP selective exosome capture sandwiched with an aptamer/graphene oxide fluorescence resonance energy transfer system (FRET) based selective 'turn-on' exosome labeling heterogeneously. The overall strategy performance was successively evaluated using lysozyme and exosomes as targets. Good linearity and high sensitivity achieved were demonstrated. The LOD of exosomal detection in serum was 2.43 × 106 particles/mL, lower than other immunology based detection methods. The discrimination between serum from breast cancer patients and healthy people was also primarily studied. In conclusion, the developed sensor with outstanding selectivity, high detection sensitivity, simplicity, low cost, and wide applicability for known or unknown targets present significant potential in challenging clinical diagnosis.
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Técnicas Biosensibles , Exosomas , Impresión Molecular , Técnicas Biosensibles/métodos , Transferencia Resonante de Energía de Fluorescencia/métodos , Grafito , Humanos , Fenómenos Magnéticos , Oligonucleótidos , PolímerosRESUMEN
Extracellular vesicles (EVs) are hemispherical vesicles that have a lipid bilayer. Studies have shown that EVs have important biological functions. The amount, types, and compositional changes of proteins, lipids and ribonucleic acids are closely related to diseases. The separation and capture of EVs from the complicated body fluid samples is a prerequisite for medical research and liquid biopsy based on EVs. However, presently the majority of EVs separation and capture still use the traditional separation methods with low purity and low efficiency. Therefore, efficient and highly selective EVs separation method is in urgent need. To meet this challenge, advanced microfluidic chip technology, which has the advantages of miniaturization, integration, and automation, can be utilized. The development of EV separation technology combined with microfluidic chips has become the focus of research. This paper summarizes the latest research progress in this area.
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Vesículas Extracelulares , Microfluídica/tendenciasRESUMEN
This article reviews the recent advances in microfluidic-chip integrated optical biosensors for simultaneous detection of multiple analytes. In particular, the principles and recent progress in different kinds of multiplex optical biosensors and their biological application were reviewed comprehensively. Sensors based on multiplexed detection have absolute advantages in analysis throughput than single assay. The microfluidic chip, a type of micro-total analysis system (µTAS), provides an ideal platform for integration of high-throughput biosensors. Compared with electronic biosensors, benefitted from the technical development in Micro-Electro-Mechanical System, there have been greater advances in the fabrication of optical sensors and microfluidic chip, and then promoting microfluidic-chip integrated optical biosensors for simultaneous detection of multiple analytes.
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Técnicas Biosensibles/métodos , Técnicas Analíticas Microfluídicas/métodos , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Técnicas Biosensibles/tendencias , Humanos , Dispositivos Laboratorio en un Chip , Sistemas Microelectromecánicos/métodos , Técnicas Analíticas Microfluídicas/tendenciasRESUMEN
Utilizing biosensors for multiplexed detection can greatly increase analysis throughput and thus, the amount of information obtained in a single assay. The microfluidic chip, a type of micro-total analysis system (µTAS), has provided a necessary platform for portable and high-throughput biosensors. Biosensors and microfluidic chips are powerful individually, and their super combination is very meaningful for analytical especially for biological applications. In this paper, every kind of microfluidic-chip-integrated electronic biosensors including some emerging technologies for simultaneous detection of multiple analytes are reviewed. Different ways to reduce or avoid cross-talking and more efforts to achieve lab on chip multisensors were also introduced to help readers form a general idea of current developments in different angles.