RESUMO
Visualizing biomolecules such as enzymes in the deep tissue of living organisms via molecular ratiometric fluorescent probes in the second near-infrared window (NIR-II) with a built-in self-calibration function can provide reliable information about relevant pathophysiological processes directly but so far is not feasible due to the lack of a fluorescence modulation strategy in the NIR-II window. Here we present a molecular platform Py-2 by integrating the rhodamine 6G scaffold and polymethine. The maximal emission wavelength of Py-2 was 1010 nm and blue-shifted to 945 nm when its secondary amine was acylated. Based on Py-2, two molecular ratiometric NIR-II fluorescent probes, nitroreductase-responsive Rap-N and ROS-responsive Rap-R, were constructed and successfully demonstrated in vitro and in vivo. Overall, this report presents a unique approach to developing ratiometric NIR-II molecular probes for in vivo biosensing.
Assuntos
Corantes Fluorescentes , Indóis , FluorescênciaRESUMO
Despite having the potential to synthesize stable metal-organic frameworks (MOFs), rare earth metal-based MOFs have not been exploited extensively. Owing to the high coordination numbers, the MOFs can generate a suitable coordination environment for various applications. Herein, samarium (Sm)-based MOFs were synthesized with three different organic linkers, namely, trimesic acid (TMA), meso-tetra(4-carboxyphenyl)porphine (TCPP), and 1,3,6,8-tetra(4-carboxylphenyl) pyrene(TBPy) by the solvothermal approach. The morphologies of Sm-TMA MOF, Sm-TCPP MOF, Sm-TBPy MOF were rod-shaped, cubic consisting of stacked 2D layers, and spherical made of small cubic structures, respectively. After the electrochemical properties of the synthesized MOFs were investigated, the MOFs were used to fabricate immunosensors for detection of carcinoembryonic antigen using a label-free signaling strategy. The immunosensors exhibited a wide linear detection range and a lower detection limit. The exhibited reproducibility and selectivity of the immunosensors were within the tolerable limits. The established label-free immunosensor has been successfully applied for detection of carcinoembryonic antigen in human serum samples, demonstrating that the rare earth metal-based MOFs are promising for construction of biosensors for medical diagnosis.
Assuntos
Técnicas Biossensoriais , Neoplasias , Biomarcadores Tumorais , Colo , Técnicas Eletroquímicas , Humanos , Imunoensaio , Limite de Detecção , Reprodutibilidade dos Testes , SamárioRESUMO
Three-dimensional Cu@Cu2O aerogels with excellent electrocatalytic activity were prepared and used as electrode matrix for constructing novel electrochemical glucose sensors. The aerogels were obtained by adding a fresh solution of NaBH4 into a mixture of CuCl2 and NaOH aqueous solutions under stirring at room temperature. The aerogels were assembled with Cu or Cu2O nanoparticles. The materials show superfine spongy-like structures with large surface-to-volume ratio, numerous active sites and good solubility. The Cu@Cu2O aerogels show highly efficient electrochemical activity toward glucose oxidation with a relatively low-onset potential (0.25 V) in 0.1 M NaOH solution. This non-enzymatic glucose sensor offers a low detection limit of 0.6 µM (S/N = 3), a high sensitivity (195 mA M-1 cm-2), and two wide linear ranges (0.001-5.2 mM, 5.2-17.1 mM) at a working voltage of 0.6 V (vs. Ag/AgCl) in alkaline solution. While in neutral pH values, the respective data are a linear analytical range from 0.1 to 10 mM; a detection limit of 54 µM (S/N = 3) and a sensitivity of 12 mA M-1 cm-2 at scan rate of 100 mV s-1. The sensor possesses high selectivity, good reproducibility and long-time stability. It was utilized to determine glucose levels in (spiked) human serum samples, and satisfactory results were obtained. Graphical abstract Schematic presentation of a glassy carbon electrode modified with 3D porous Cu@Cu2O aerogels. The aerogels were obtained by a reduction reaction at room temperature (Scheme 1A). The aerogel networks were used to develop a highly sensitive electrochemical sensing platform for the detection of glucose (Scheme 1B).
Assuntos
Glicemia/análise , Cobre/química , Nanopartículas Metálicas/química , Técnicas Biossensoriais , Catálise , Técnicas Eletroquímicas , Eletrodos , Géis/química , Humanos , Limite de Detecção , Oxirredução , Tamanho da Partícula , Porosidade , Reprodutibilidade dos Testes , Propriedades de SuperfícieRESUMO
BACKGROUND: Hypochlorous acid (HClO) is an important biomarker for inflammation, cardiovascular disease, and even cancer. It is of great significance to accurately monitor and quantitatively analyze the fluctuations of HClO to better understand their physiological functions. Traditional HClO detection methods such as high-performance liquid chromatography (HPLC), and mass spectrometry are preferred, but are costly and unsuitable in vivo. Near-infrared (NIR) fluorescence imaging has the advantages of high sensitivity, high temporal and spatial resolutions, minimal autofluorescence, and deep tissue penetration, which facilitates its application in biological systems. Therefore, the development of sensitivity and simple NIR fluorescence monitoring HClO methods in vivo and in vitro is essential and desirable. RESULTS: Herein, we present a NIR probe NOF3 by integrating the rhodamine scaffold and HClO-triggered moiety for the real-time detection of HClO in vitro and in vivo. NOF3 reacts with the HClO and releases the NOF-OH fluorophore of emitted signals at 730 nm, which is in the NIR region. The designed probe detected concentrations of HClO ranging from 0 to 17 µM with a low detection limit of 0.146 µM, presenting excellent sensitivity and selectivity toward HClO over other species. NOF3 manifests significantly turn-on NIR fluorescent signals in response to HClO concentration, which makes it favorable for monitoring dynamic HClO distribution in vivo. We exemplify NOF3 for the tracking of endogenously overexpressed HClO distribution in RAW 264.7 cells, and further realize real-time in vivo bioimaging of HClO activity in inflammation mice. SIGNIFICANCE: The facile NIR NOF3 probe was successfully applied to visualize endogenous and exogenous HClO in living cells and mice. This study provides not only an effective tool for spatial and temporal resolution HClO bioimaging in vivo but also possesses great potential for use in future research on HClO-related biology and pathology.
Assuntos
Ácido Hipocloroso , Xantenos , Camundongos , Animais , Ácido Hipocloroso/análise , Rodaminas/química , Corantes Fluorescentes/química , Inflamação/diagnóstico por imagemRESUMO
A novel biothiols-sensitive near-infrared (NIR) fluorescent probe RhDN based on a rhodamine skeleton was developed for early detection of drug-induced hepatotoxicity in living mice. RhDN can be used not only as a conventional large stokes shift fluorescent (FL) probe, but also as a kind of anti-Stokes frequency upconversion luminescence (FUCL) molecular probe, which represents a long wavelength excitation (808 nm) to short wavelength emission (760 nm), and response to Cys/Hcy/GSH with high sensitivity. Compared with traditional FL methods, the FUCL method exhibited a lower detection limit of Cys, Hcy, and GSH in 75.1 nM, 101.8 nM, and 84.9 nM, respectively. We exemplify RhDN for tracking endogenously biothiols distribution in living cells and further realize real-time in vivo bioimaging of biothiols activity in mice with dual-mode luminescence system. Moreover, RhDN has been successfully applied to visualize the detection of drug-induced hepatotoxicity in living mice. Overall, this report presents a unique approach to the development of large stokes shift NIR FUCL molecular probes for in vitro and in vivo biothiols biosensing.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Corantes Fluorescentes , Animais , Corantes Fluorescentes/química , Corantes Fluorescentes/toxicidade , Doença Hepática Induzida por Substâncias e Drogas/diagnóstico por imagem , Camundongos , Humanos , Raios Infravermelhos , Imagem Óptica , Glutationa/análise , Compostos de Sulfidrila/análise , Compostos de Sulfidrila/química , Cisteína/análise , Rodaminas/química , Rodaminas/toxicidade , Homocisteína/análise , LuminescênciaRESUMO
In this work, a nanocomposite of Zr-trimesic acid MOF (MOF-808) with carbon nanotube (CNT) was synthesized through an in situ formation of MOF-808 on the activated CNT. The synthesized materials were characterized by powder X-ray diffraction, transmission electron microscopy, X-ray photoluminescence spectroscopy, Brunauer-Emmett-Teller, Fourier transform infrared spectroscopy, and Raman spectroscopy. The protein compatible nature with high surface area and electrocatalytic ability of MOF-808 was utilized to construct an immunosensor for ultra low-level detection of the ovarian cancer biomarker, carbohydrate antigen 125 (CA 125). The mutual benefit of each constituent of the MOF-808/CNT composite was capable of producing highly enhanced electrochemical properties. A glassy carbon electrode modified with MOF-808/CNT was used as a platform to fabricate a label-free electrochemical immunosensor. The antibody binding sites of MOF-808/CNT were enriched by functionalization with streptavidin. The immunosensor exhibited two linear determination ranges of 0.001-0.1 and 0.1-30 ng·mL-1, and the calculated limit of detection was 0.5 pg·mL-1 (S/N 3). The immunosensor showed excellent reproducibility and selectivity. The patient serum sample analysis was cross-verified with the electrochemiluminescence method with a relative error of 105-110%.
Assuntos
Anticorpos Imobilizados/química , Técnicas Biossensoriais/métodos , Antígeno Ca-125/sangue , Estruturas Metalorgânicas/química , Nanotubos de Carbono/química , Técnicas Eletroquímicas/métodos , Eletrodos , Humanos , Imunoensaio/métodos , Limite de Detecção , Neoplasias/sangueRESUMO
In this work, a magnetic graphene oxide-ultrathin metal-organic framework composite (Fe3O4@SiO2-GO-Ni-MOF) was synthesized for the first time. Employing Fe3O4@SiO2-GO-Ni-MOF composite as extractant, a novel method for the separation and analysis of the pesticide epoxiconazole was established with the assistance of high performance liquid chromatography (HPLC). The adsorption mechanisms were studied including by adsorption kinetics, thermodynamic parameters and adsorption isotherms. The experimental results showed that this method was convenient, operable, effective and practical for the extraction and determination of epoxiconazole in real samples.
RESUMO
Herein, we report the synthesis of novel nanosized reduced graphene oxide@polystyrene nanospheres (rGO@PS NSs) through an electrostatic interaction method, and further their exploitation for highly sensitive label-free electrochemical immunoassay applications. These rGO@PS NSs provide a universal and promising carrier platform to develop excellent biosensors for clinical screening and diagnostic applications.
Assuntos
Biomarcadores Tumorais/análise , Grafite/química , Nanopartículas/química , Poliestirenos/química , alfa-Fetoproteínas/análise , Animais , Anticorpos Monoclonais Murinos/imunologia , Biomarcadores Tumorais/imunologia , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Imunoensaio/métodos , Limite de Detecção , Camundongos , Reprodutibilidade dos Testes , alfa-Fetoproteínas/imunologiaRESUMO
Quantitative detection of multiple chicken cytokines is a good evaluation of cell-mediated immunity in chickens after disease infection or vaccination. However, current assay methods for chicken cytokines cannot meet the needs of clinical diagnosis due to unsatisfactory sensitivity and low assay throughput. Herein, a sensitive chemiluminescence (CL) imaging immunosensor array has been developed for high-throughput detection of multiple chicken cytokines. The chicken cytokines immunosensor array was prepared by assembling different cytokine capture antibodies onto a disposable silanized glass chip, where horseradish peroxidase and antibody-conjugated gold nanoparticles were used as multienzymatic amplification probe for CL imaging signal amplification. By using a sandwich assay mode, the amplified CL signals from each sensing array cell were collected for quantitation. Using chicken interleukin-4 and chicken interferon-γ as model cytokines, this novel multiplexed and amplified method demonstrated simultaneous measurement of the two chicken cytokines in the linear ranges of 0.008-0.12â¯ng/mL and 0.005-0.20â¯ng/mL, respectively, which yields limits of detection down to 2â¯pg/mL and 3â¯pg/mL. The CL imaging array method reported here also demonstrated high specificity, good repeatability, and high stability and accuracy, providing a novel multiplex immunoassay strategy for highly sensitive and high-throughput detection of chicken cytokines and further disease diagnosis in poultry.
Assuntos
Citocinas/sangue , Imunoensaio/métodos , Animais , Anticorpos Imobilizados/imunologia , Galinhas , Citocinas/imunologia , Ouro/química , Peroxidase do Rábano Silvestre/química , Limite de Detecção , Luminescência , Medições Luminescentes/métodos , Nanopartículas Metálicas/química , Reprodutibilidade dos TestesRESUMO
We report a new concept of a chemiluminescence imaging nanozyme immunoassay (CINIA), in which nanozymes are exploited as catalytic tags for simultaneous multiplex detection of cytokines. The CINIA provides a novel and universal nanozyme-labeled multiplex immunoassay strategy for high-throughput detection of relevant biomarkers and further disease diagnosis.
Assuntos
Citocinas/análise , Medições Luminescentes/métodos , Animais , Anticorpos Imobilizados/química , Galinhas , Cobre/química , Citocinas/sangue , Desenho de Equipamento , Ensaios de Triagem em Larga Escala/instrumentação , Ensaios de Triagem em Larga Escala/métodos , Imunoensaio/instrumentação , Imunoensaio/métodos , Medições Luminescentes/instrumentação , Nanopartículas/química , Sulfetos/químicaRESUMO
In this work, an efficient and universal streptavidin-functionalized nitrogen-doped graphene (NG) was for the first time proposed and used to develop a highly sensitive electrochemical immunosensor for the detection of tumor markers. Transmission electron microscopy, electrochemical impedance spectrum, static water contact measurement, and cyclic voltammetry were used to characterize the streptavidin-functionalized NG platform and immunosensor. The biofunctionalized NG showed excellent hydrophilicity, larger specific surface area, and high electrochemical activity. These properties of the platform enhanced the loading capacity of proteins, and retained the bioactivity of the immobilized proteins, and thus remarkably improved the sensitivity of the immunosensor. Using carcinoembryonic antigen (CEA) as model analyte, the proposed immunosensor demonstrated a wide linear range of 0.02-12ngmL-1 with a low detection limit of 0.01ngmL-1. The CEA immunosensor could be applied to detect human serum samples with satisfactory results. The streptavidin-functionalized NG material provided an universal and promising platform for the electrochemical immunosensing applications.