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1.
Biosens Bioelectron ; 261: 116492, 2024 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-38870828

RESUMO

Exosomes have been considered as promising biomarkers for cancer diagnosis due to their abundant information from originating cells. However, sensitive and reliable detection of exosomes is still facing technically challenges due to the lack of a sensing platform with high sensitivity and reproducibility. To address the challenges, here we propose a portable surface plasmon resonance (SPR) sensing of exosomes with a three-layer Au mirror/SiO2 spacer/Au nanohole sensor, fabricated by an economical polystyrene nanosphere self-assembly method. The SiO2 spacer can act as an optical cavity and induce mode hybridization, leading to excellent optimization of both sensitivity and full width at half maximum compared with normal single layer Au nanohole sensors. When modified with CD63 or EpCAM aptamers, a detection of limit (LOD) of as low as 600 particles/µL was achieved. The sensors showed good capability to distinguish between non-tumor derived L02 exosomes and tumor derived HepG2 exosomes. Additionally, high reproducibility was also achieved in detection of artificial serum samples with RSD as low as 2%, making it feasible for clinical applications. This mode hybridization plasmonic sensor provides an effective approach to optimize the detection sensitivity of exosomes, pushing SPR sensing one step further towards cancer diagnosis.

2.
Biomater Adv ; 161: 213891, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38781738

RESUMO

An antitumour chemo-photodynamic therapy nanoplatform was constructed based on phospholipid-coated NaYF4: Yb/Er upconversion nanoparticles (UCNPs). In this work, the amphiphilic block copolymer DSPE-PEG2000 was combined with the surface ligand oleic acid of the UCNPs through hydrophobic interaction to form liposomes with a dense hydrophobic layer in which the photosensitizer hypocrellin B (HB) was assembled. The coated HB formed J-aggregates, which caused a large redshift in the absorption spectrum and improved the quantum efficiency of energy transfer. Furthermore, MnO2 nanosheets grew in-situ on the liposomes through OMn coordination. Therefore, a multifunctional tumour microenvironment (TME)-responsive theranostic nanoplatform integrating photodynamic therapy (PDT) and chemodynamic therapy (CDT) was successfully developed. The results showed that this NIR-mediated chemo-photodynamic therapy nanoplatform was highly efficient for oncotherapy.


Assuntos
Compostos de Manganês , Nanopartículas , Óxidos , Perileno , Fotoquimioterapia , Fármacos Fotossensibilizantes , Quinonas , Fotoquimioterapia/métodos , Perileno/análogos & derivados , Perileno/farmacologia , Perileno/química , Perileno/administração & dosagem , Humanos , Quinonas/química , Quinonas/farmacologia , Nanopartículas/química , Nanopartículas/uso terapêutico , Óxidos/química , Óxidos/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/uso terapêutico , Fármacos Fotossensibilizantes/administração & dosagem , Compostos de Manganês/química , Compostos de Manganês/farmacologia , Animais , Fenol/química , Fenol/farmacologia , Lipossomos/química , Antineoplásicos/farmacologia , Antineoplásicos/química , Antineoplásicos/administração & dosagem , Antineoplásicos/uso terapêutico , Camundongos , Linhagem Celular Tumoral , Microambiente Tumoral/efeitos dos fármacos
3.
Mikrochim Acta ; 191(1): 15, 2023 12 13.
Artigo em Inglês | MEDLINE | ID: mdl-38087000

RESUMO

Based on upconversion nanoparticles (UCNPs) as energy donor and herring sperm DNA (hsDNA) as molecular recognition element, an unlabelled upconversion luminescence (UCL) affinity biosensor was constructed for the detection of anthraquinone (AQ) anticancer drugs in biological fluids. AQ anticancer drugs can insert into the double helix structure of hsDNA on the surface of UCNPs, thereby shortening the distance from UCNPs. Therefore, the luminescence resonance energy transfer (LRET) phenomenon is effectively triggered between UCNPs and AQ anticancer drugs. Hence, AQ anticancer drugs can be quantitatively detected according to the UCL quenching rate. The biosensor showed good sensitivity and stability for the detection of daunorubicin (DNR) and doxorubicin (ADM). For the detection of DNR, the linear range is 1-100 µg·mL-1 with a limit of detection (LOD) of 0.60 µg·mL-1, and for ADM, the linear range is 0.5-100 µg·mL-1 with a LOD of 0.38 µg·mL-1. The proposed biosensor provides a convenient method for monitoring AQ anticancer drugs in clinical biological fluids in the future.


Assuntos
Antineoplásicos , Técnicas Biossensoriais , Masculino , Humanos , Sêmen , DNA , Técnicas Biossensoriais/métodos , Antraquinonas
4.
Anal Bioanal Chem ; 415(29-30): 7139-7150, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37803135

RESUMO

In this work, an upconversion luminescence (UCL) nanosensor for fast detection of ferric ion (Fe3+) and phosphate ion (Pi) is developed based on the inner-filter effect (IFE) between NaYF4:Yb/Er upconversion nanoparticles (UCNPs) and Fe3+-hypocrellin B (HB) complex. Fe3+-HB complex has strong absorption band (450-650 nm), which overlaps with the green emission peak of UCNPs at 545 nm. By adding Fe3+ and Pi, the UCNPs-HB system produces the red-shift change of absorption spectrum, which leads to the "on-off-on" process of IFE. So, with the specific recognition ability of HB for Fe3+ and the competitive complexation of Pi for Fe3+, the proposed nanosensor utilizes the UCL change to achieve the detection of the targets. For the detections of Fe3+, the linear range is 10-600 µM with a limit of detection (LOD) of 2.62 µM, and for Pi, the linear range is 5-100 µM with a LOD of 1.25 µM. The results for selectivity, precision, and recovery test are also satisfactory. Furthermore, the real sample detection shows that the proposed nanaosensor has a great potential in environmental and biological systems. An upconversion luminescence (UCL) nanosensor based on the inner-filter effect (IFE) between upconversion nanoparticles (UCNPs) and Fe3+-hypocrellin B (HB) complex for the detection of Fe3+ and phosphate ion has been proposed, which is promising to be a convenient and sensitive assay for monitoring Fe3+ and phosphate ion in different environments and biological systems.

5.
Front Chem ; 10: 1028441, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36267653

RESUMO

Based on the mechanism of luminescence resonance energy transfer (LRET) and using a special single strand DNA as the recognition element, a portable paper-based sensor for the accurate detection of total heavy rare-earth ions (mainly Gd3+, Tb3+ and Dy3+) concentration was proposed. The RNA cleaving-DNAzyme should recognize rare-earth ions to cleave RNA on DNA duplexes linking UCNPs and AuNPs, causing UCNPs and AuNPs to approach each other, inducing LRET, which attenuated the green upconversion luminescence (UCL) triggered by the 980 nm laser. UCL was captured by a charge-coupled device (CCD) image sensor and processed with the red-green-blue (RGB) image to quantitatively analyze heavy rare-earth ions in the samples. In the range of 5-50 µmol·L-1, the sensor has good sensitivity, with the limit of detection of 1.26 µmol L-1.

6.
ACS Appl Mater Interfaces ; 14(24): 27686-27702, 2022 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-35675505

RESUMO

To overcome the low efficacy of conventional monotherapeutic approaches that use a single drug, functional nanocarriers loaded with an amalgamation of anticancer drugs have been promising in cancer therapy. Herein, aloe-derived nanovesicles (gADNVs) are modified with an active integrin-targeted peptide (Arg-Gly-Asp, RGD) by the postinsertion technique to deliver indocyanine green (ICG) and doxorubicin (DOX) for efficient breast cancer therapy. We presented for the first time that the π-π stacking interaction can turn the "competitive" relationship of ICG and DOX inside gADNVs into a "cooperative" relationship and enhance their loading efficiency. The dual-drug codelivery nanosystem, denoted as DIARs, was well stable and leakproof, exhibiting high tumor-targeting capability both in vitro and in vivo. Meanwhile, this nanosystem showed significant inhibition of cell growth and migration and induced cell apoptosis with the combination of phototherapy and chemotherapy. Intravenous administration of DIARs exhibited high therapeutic efficacy in a 4T1 tumor-bearing mouse model and exhibited no obvious damage to other organs. Overall, our DIAR nanosystem constitutively integrated the natural and economical gADNVs, π-π stacking interaction based on efficient drug loading, and tumor-targeted RGD modification to achieve an effective combination therapy for breast cancer.


Assuntos
Aloe , Antineoplásicos , Nanopartículas , Neoplasias , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Doxorrubicina/uso terapêutico , Verde de Indocianina/farmacologia , Camundongos , Neoplasias/tratamento farmacológico , Oligopeptídeos
7.
Biosens Bioelectron ; 209: 114259, 2022 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-35421672

RESUMO

The analysis of microRNAs (miRNAs) in exosomes offers significant information for a rapid and non-invasive diagnosis of cancer. However, the clinical utility of miRNAs as biomarkers is often hampered by their low abundance in exosomes. Herein, we develop a dual-signal amplification biosensor for the sensitive detection of exosomal miRNA-21 (miR-21). In the presence of a cognate target, it hybridizes with a biotin-modified capture probe (Cp) to form a DNA-RNA heteroduplex that serves as a substrate for duplex-specific nuclease (DSN). With the assistance of DSN, the Cps are enzymatically hydrolyzed and numerous DNA catalysts are released, leading to the first signal amplification. After magnetic isolation, the DNA catalyst remaining in the supernatant triggers a strand displacement reaction based on the nicking-assisted reactant recycling strategy, without depleting the reactants, to implement the second signal amplification. Using this dual-signal amplification concept, our biosensor achieves a limit of detection of miR-21 of 0.34 fM, with a linear range of 0.5-100 fM. The receiver operating characteristic curve generated during clinical sample analysis indicates that the exosomal miR-21 outperforms serum carcinoembryonic antigen in discriminating between patients with gastric cancer (GC) and patients with precancerous (PC) lesions (area under the curve: 0.89 versus 0.74, n = 40). Moreover, the proposed biosensor exhibits an 83.9% accuracy in classifying patients with GC or PC lesions and healthy donors using a confusion matrix. Furthermore, patients with GC with or without metastases are discriminated using the proposed biosensor. Our technology may expand the applications of DNA-based biosensor-enabled cancer diagnostic tools.


Assuntos
Técnicas Biossensoriais , Exossomos , MicroRNAs , Neoplasias Gástricas , Corantes , DNA , Endonucleases , Exossomos/química , Exossomos/genética , Humanos , Limite de Detecção , MicroRNAs/análise , Técnicas de Amplificação de Ácido Nucleico , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/genética
8.
Mikrochim Acta ; 189(5): 179, 2022 04 07.
Artigo em Inglês | MEDLINE | ID: mdl-35386003

RESUMO

Thyroid-stimulating hormone (TSH) plays a crucial physiological and pathological role in humans, and a timely and sensitive detection of TSH is critical for early diagnosis and prevention of thyroid-related diseases. Herein, we developed a simple wash-free biological aptasensor based on luminescence resonance energy transfer (LRET) between NaYF4:Yb,Er upconversion nanoparticles (UCNPs) and tetramethylrhodamine (TAMRA) for the detection of TSH with high sensitivity. In this LRET system, UCNPs as donors and TAMRA as receptors were modified with nucleic acid aptamers Apt-1 and Apt-2, respectively. When TSH was present, the two aptamer strands both specifically recognized TSH to form a hairpin-like structure, thereby shortening the space between UCNPs and TAMRA. The LRET occurred under radiation of 980-nm light. By detecting the change of upconversion luminescence (UCL) intensity (I545nm), the activity of TSH was quantified. The resulting detection dynamic range and the limit of detection were 0.1-5.0 mIU·L-1 and 0.065 mIU·L-1, respectively. The aptasensor using UCNPs as LRET donors was capable of effectively eliminating the background interference of a complicated biological environment, and showed good specificity because of the excellent recognition function of aptamers. Due to high sensitivity, easiness of fabrication, operational convenience, and selectivity, the UCL-based aptasensor is a promising candidate for clinical TSH determination. Based on nucleic acid aptamer and the mechanism of luminescence resonance energy transfer (LRET) between upconversion nanoparticles (UCNPs) donor and tetramethylrhodamine (TAMRA) receptor, an aptasensor was constructed for the quantitative analysis of TSH activity in serum by testing the change of I545nm.


Assuntos
Luminescência , Ácidos Nucleicos , Transferência Ressonante de Energia de Fluorescência/métodos , Humanos , Limite de Detecção , Tireotropina
9.
Anal Chem ; 94(13): 5382-5391, 2022 04 05.
Artigo em Inglês | MEDLINE | ID: mdl-35323012

RESUMO

It remains a huge challenge to integrate the sensitivity, stability, reproducibility, and anti-fouling ability of electrochemical biosensors for practical applications. Herein, we propose a self-assembled electrode combining hexanethiol (HT), poly-adenine (poly-A), and cholesteryl-modified DNA to meet this challenge. HT can tightly pack at the electrode interface to form a hydrophobic self-assembled monolayer (SAM), effectively improving the stability and signal-to-noise ratio (SNR) of electrochemical detection. Cholesteryl-modified DNA was immobilized at the electrode through the hydrophobic interaction with HT to avoid the competition between the SAM and the DNA probe on the gold site. Thus, the assembly efficiency and uniformity of the DNA probe as well as the detection reproducibility were increased remarkedly. Poly-A was added on the HT assembled electrode to occupy the unreacted sites of gold to further enhance the anti-fouling ability. The combination of HT and poly-A allows the electrode to ensure favorable anti-fouling ability without sacrificing the detection performance. On this basis, we proposed a dual-signal amplification electrochemical biosensor for the detection of exosomal microRNAs, which showed excellent sensitivity with a detection limit down to 1.46 aM. Importantly, this method has been successfully applied to detect exosomal microRNA-21 in cells and human serum samples, proving its potential utility in cancer diagnosis.


Assuntos
Incrustação Biológica , Técnicas Biossensoriais , MicroRNAs , Incrustação Biológica/prevenção & controle , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Eletrodos , Ouro , Humanos , Limite de Detecção , MicroRNAs/genética , Reprodutibilidade dos Testes
10.
Anal Chim Acta ; 1191: 339279, 2022 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-35033266

RESUMO

Exosomes are promising biomarkers for cancer screening, but the development of a robust approach that can sensitively and accurately detect exosomes remains challenging. In the present study, an aptasensor based on the multifunctional signal probe 10-benzyl-2-amino-acridone (BAA) was developed for the colorimetric and photoelectrochemical detection and quantitation of exosomes. Exosomes are captured by cholesterol DNA anchor-modified magnetic beads (MBs) through hydrophobic interactions. This capture process can be monitored under a confocal fluorescence microscope using BAA as the fluorescent signal probe. The aptamer modified copper oxide nanoparticles (CuO NPs) then bind to mucin 1 (MUC1) on the surface of the exosomes to form a sandwich structure (MBs-Exo-CuO NPs). Finally, the MBs-Exo-CuO NPs are dissolved in nitric acid to generate Cu2+, which inhibits the visible-light-induced oxidase mimic activity and photoelectrochemical activity of BAA simultaneously. The changes in absorbance and photocurrent intensities are directly proportional to the concentration of exosomes. In this dual-modal aptasensor, the colorimetric assay can achieve rapid screening and identification, which is especially useful for point-of-care testing. The UV-vis absorbance and photocurrent assays then provide quantitative information, with a limit of detection of 1.09 × 103 particles µL-1 and 1.38 × 103 particles µL-1, respectively. The proposed aptasensor thus performs dual-modal detection and quantitation of exosomes. This aptasensor provides a much-needed toolset for exploring the biological roles of exosomes in specific diseases, particularly in the clinical setting.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Exossomos , Acridonas , Colorimetria , Limite de Detecção
11.
J Nanobiotechnology ; 19(1): 439, 2021 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-34930289

RESUMO

BACKGROUND: Cancer is one of the devastating diseases in the world. The development of nanocarrier provides a promising perspective for improving cancer therapeutic efficacy. However, the issues with potential toxicity, quantity production, and excessive costs limit their further applications in clinical practice. RESULTS: Herein, we proposed a nanocarrier obtained from aloe with stability and leak-proofness. We isolated nanovesicles from the gel and rind of aloe (gADNVs and rADNVs) with higher quality and yield by controlling the final centrifugation time within 20 min, and modulating the viscosity at 2.98 mPa S and 1.57 mPa S respectively. The gADNVs showed great structure and storage stability, antioxidant and antidetergent capacity. They could be efficiently taken up by melanoma cells, and with no toxicity in vitro or in vivo. Indocyanine green (ICG) loaded in gADNVs (ICG/gADNVs) showed great stability in both heating system and in serum, and its retention rate exceeded 90% after 30 days stored in gADNVs. ICG/gADNVs stored 30 days could still effectively damage melanoma cells and inhibit melanoma growth, outperforming free ICG and ICG liposomes. Interestingly, gADNVs showed prominent penetrability to mice skin which might be beneficial to noninvasive transdermal administration. CONCLUSIONS: Our research was designed to simplify the preparation of drug carrier, and reduce production cost, which provided an alternative for the development of economic and safe drug delivery system.


Assuntos
Aloe/química , Verde de Indocianina/química , Nanoestruturas/química , Aloe/metabolismo , Animais , Antioxidantes/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Citocinas/sangue , Estabilidade de Medicamentos , Hemólise/efeitos dos fármacos , Humanos , Verde de Indocianina/farmacologia , Verde de Indocianina/uso terapêutico , Lipossomos/química , Melanoma Experimental/tratamento farmacológico , Camundongos , Nanoestruturas/uso terapêutico , Nanoestruturas/toxicidade , Tamanho da Partícula
12.
Anal Chim Acta ; 1143: 37-44, 2021 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-33384128

RESUMO

Phospholipase A2 (PLA2) may be a vital biomarker for the prediction and diagnosis of some diseases. Consequently, it is of great significance to quantitatively detect PLA2 in biologic samples. Herein, on the basis of the principle of luminescence resonance energy transfer (LRET) between upconversion nanoparticles (UCNPs) and SYBR Green I (SG), we proposed a technology for the highly sensitive detection of PLA2 amount. Therein, as an energy receptor, SG will be quantitatively loaded into liposomes firstly. Then, due to the hydrolysis of liposomes under the catalysis of PLA2, SG will be released and inserted into the double-stranded DNA (dsDNA) on the surface of UCNPs, which triggers the LRET because of the shortening of effective spatial distance between UCNPs and SG. Under exciting of NIR light, UCNPs emit luminescence at 476 nm, which makes SG emit fluorescence at 522 nm through LRET. Under optimal conditions, the emission intensity ratio (I522 nm/I476 nm) increased linearly with the PLA2 amount in the range of 20 U/L to 400 U/L, and the limit of detection (LOD) reached 15 U/L. Here, after comparing with the clinical standard method, it is found that the biosensor is expected to provide a convenient and sensitive assay for the detection of PLA2 in actual serum samples. Furthermore, such biosensor can also be used to test the inhibitor of PLA2.


Assuntos
Transferência Ressonante de Energia de Fluorescência , Nanopartículas , Benzotiazóis , Diaminas , Compostos Orgânicos , Fosfolipases , Quinolinas
13.
Biosens Bioelectron ; 173: 112834, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33243697

RESUMO

Exosomal microRNAs (miRNAs) are vital biomarkers for early diagnosis and prognosis monitoring of cancer. Yet, convenient and controllable detection of exosomal miRNA still remains challenges because of lacking of adequately simple and robust assay platforms. In this paper, it is first time to study the visible-light-induced oxidase mimic activity of 10-methyl-2-amino-acridone (MAA) being able to be switched by Cu2+ and DNA. Based on this phenomenon, a series of visual molecular logic gates are constructed, and a colorimetric strategy has been developed to achieve exosomal microRNA-21 (miR-21) detection with a signal amplification approach. The visible-light-induced oxidase mimic activity of MAA can be inhibited by Cu2+. In presence of target, a large amount of capture probes partly complementary with miR-21 are hydrolyzed with the assist of duplex-strand specific nuclease (DSN), releasing guanine-rich oligodeoxynucleotides that can chelate Cu2+, resulting in catalytic activity of MAA being recovered under irradiation. This strategy allows the detection of miR-21 with a light modulating temporal controllable manner, and the linear range is from 50 fM to 3000 fM with the limit of detection (LOD) being 44.76 fM. More importantly, the proposed method can achieve quantitative measurement of exosomal miR-21 that is derived from three-dimensional multicellular tumor spheroids with different size, which is able to monitor the growth of tumor spheroids. This work is potential to provide a feasible tool for application in exosomal miRNAs-based cancer diagnosis. Ultimately, MAA is expected to be a signal probe in biomedical field by virtue of its fascinating visible-light-induced oxidase mimic activity.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Acridonas , Colorimetria , MicroRNAs/genética , Oxirredutases
14.
Talanta ; 214: 120851, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32278412

RESUMO

As non-invasive biomarkers, exosomes are of great significance to diseases diagnosis. However, sensitive and accurate detection of exosomes still remains technical challenges. Herein, inspired by nature's "one-to-many" concept, we design a biosensor mimicking the cactus with numerous thorns to detect exosomes. The biosensor is composed of CD63 antibodies, resembling the roots of cactus, to capture exosomes, and the exosomes resemble the stems. Cholesterol-labeled DNA (DNA anchor) binding to streptavidin modified horseradish peroxidase (HRP) can insert into exosomes membrane, which seems the thorns. The readout signal is produced through HRP-catalyzed hydrogen peroxide (H2O2) mediated oxidation of 1,4-phenylenediamine (PPD) to form 2,5-diamino-NN'-bis-(p-aminophenyl)-1,4-benzoquinone di-imine (PPDox). The PPDox can quench fluorescence of fluorescein through inner filter effect (IFE), which provides fluorescent signal for exosomes detection. Based on this principle, the obtained exosomes solution is qualitatively and quantitatively analyzed by our biosensor, with the comparison to current standard methods by nanoparticle tracking analysis (NTA) and commercial enzyme-linked immunosorbent assay (ELISA) kit. The linear range is from 1.0 × 104 to 5.0 × 105 particles µL-1 with the limit of detection 3.40 × 103 particles µL-1 and 3.12 × 103 particles µL-1 for colorimetric and fluorescent assays, respectively. Meanwhile, our biosensor exhibits good selectivity, and can eliminate the interference from proteins. This dual-modal biosensor shows favorable performance towards analytical application in clinic samples, pushing one step further towards practical clinical use.


Assuntos
Técnicas Biossensoriais , Colorimetria , Ensaio de Imunoadsorção Enzimática , Exossomos/química , Fluorescência , Biocatálise , Biomarcadores/análise , Biomarcadores/metabolismo , Exossomos/metabolismo , Peroxidase do Rábano Silvestre/metabolismo , Humanos , Peróxido de Hidrogênio/química , Peróxido de Hidrogênio/metabolismo , Tamanho da Partícula , Fenilenodiaminas/química , Fenilenodiaminas/metabolismo , Propriedades de Superfície
15.
Talanta ; 207: 120257, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31594620

RESUMO

A label-free electrochemical biosensor based on the triplex DNA-templated Ag/Pt bimetallic nanoclusters (triplex-Ag/PtNCs) and locked nucleic acid (LNA) modified X-shaped DNA probe was developed for the detection of single-nucleotide variant (SNV) related to ß-thalassemia. Firstly, using triplex DNA as template, a site-specific and homogeneous Ag/PtNCs was prepared, which can effectively catalyze the 3,3,5,5-tetramethylbenzidine-H2O2 system and thus be employed as a signal reporter in the field of electrochemical biosensor. Secondly, the LNA modified X-shaped probes were assembled on gold electrode surface, which can only be dissociated in the presence of target, leading to the hybridization with triplex-Ag/PtNCs and significant increase of current signal. In this way, the detection limit for SNV of ß-thalassemia was 0.8 fM with variant allele frequency (VAF) as low as 0.0001%.


Assuntos
Técnicas Biossensoriais/métodos , DNA/química , Platina/química , Polimorfismo de Nucleotídeo Único , Prata/química , Catálise , Sondas de DNA/química , Sondas de DNA/metabolismo , Eletroquímica , Limite de Detecção , Nanopartículas Metálicas , Modelos Moleculares , Conformação de Ácido Nucleico , Oligonucleotídeos/química , Oligonucleotídeos/metabolismo
16.
Anal Chem ; 91(13): 8406-8414, 2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31136153

RESUMO

Compared with plenty of single-functional molecules, multifunctional molecules are scarce and have high demand in further research. In this work, a multifunctional molecule called 10-methyl-2-amino-acridone (MAA) is presented. Interestingly, MAA simultaneously features electrochemistry, two-photon fluorescence, visible-light-induced oxidase mimic, and photoelectrochemistry (PEC) activity, and the related properties are studied in detailed. Multiple functions integrated into one molecule allow MAA to become a versatile signal probe. Therefore, the MAA acted as an electrochemical indicator to detect exosomal total protein with high sensitivity at first. In addition, MAA is used for one- or two-photon fluorescence imaging in vitro and in vivo, including cells, three-dimensional (3D) tumor spheroids, zebrafish, and exosomes. The results suggest that MAA not only possesses favorable photostability, but it is also suitable for imaging in deep tissue. Furthermore, the visible-light-induced oxidase mimic and photoelectrochemical activities of MAA are selectively inhibited by Cu2+, and the relevant mechanism is carefully analyzed. On the basis of this phenomenon, we develop a dual-modal detection strategy for detection of Cu2+ in river water. Compared with a single signal readout model, this strategy is able to avoid false positive and negative detection through two series of data mutually validating each other. Therefore, our study shows that the "multiple-in-one" MAA provides a blueprint for the investigation and application of a multifunctional organic molecule.

17.
Anal Chem ; 90(15): 8969-8976, 2018 08 07.
Artigo em Inglês | MEDLINE | ID: mdl-29973048

RESUMO

Recently, sensitive and selective detection of exosomal microRNAs (miRNAs) has been garnering significant attention, because it is related to many complex diseases, including cancer. Herein, we report a ratiometric fluorescent bioprobe based on DNA-labeled carbon dots (DNA-CDs) and 5,7-dinitro-2-sulfo-acridone (DSA) coupling with the target-catalyzing signal amplification for the detection of exosomal miRNA-21. There was high fluorescence resonance energy transfer (FRET) efficiency between carbon dots (CDs) and DSA when the bioprobe was assembled. However, in the presence of the target, with disassembling of the fluorescent bioprobe, the fluorescence intensities of CDs and DSA were changed simultaneously. Because of the ratio of dual fluorescence intensities, this ratiometric fluorescent bioprobe was able to cancel out environmental fluctuations by calculating emission intensity ratio at two different wavelengths, being robust and stable enough for detection of exosomal miRNA-21. In addition, we displayed that a single miRNA-21 can catalyze the disassembly of multiple CDs with DSA theoretically, yielding significant change in the fluorescence ratio for the detection of miRNA-21. With this signal amplification strategy, the limit of detection was as low as 3.0 fM. Furthermore, because of the introduction of lock nucleic acid to mediate the strand displacement reaction, the selectivity of this strategy was improved remarkably, even against single base mismatch sequence. More importantly, our strategy could monitor the dynamic change of exosomal miRNA-21, which maybe becomes a potential tool to distinguish cancer exosomes and nontumorigenic exosomes. In a short, this ratiometric fluorescence bioprobe possessed high stability, sensitivity and selectivity coupling with ease of operation and cost efficiency, leading to great potential for wide application.


Assuntos
Acridonas/química , Carbono/química , DNA/química , Exossomos/química , Corantes Fluorescentes/química , MicroRNAs/análise , Pontos Quânticos/química , Técnicas Biossensoriais/métodos , Transferência Ressonante de Energia de Fluorescência/métodos , Humanos , Células MCF-7
18.
Biosens Bioelectron ; 102: 582-588, 2018 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-29241062

RESUMO

Exosomes, as potential cancer diagnostic markers have received close attention in recent years. However, there is still a lack of simple and convenient methods to detect and quantitate exosomes. Herein, we used a simple paper-supported aptasensor based on luminescence resonance energy transfer (LRET) from upconversion nanoparticles (UCNPs) to gold nanorods (Au NRs) for the accessible determination of exosomes. When exosomes are present, the two sections of the aptamer can combine with the CD63 protein on the surface of exosomes and form a conjugation to close the distance between UCNPs and Au NRs, which initiates the LRET and promotes luminescence quenching. These variations can be monitored by the homemade image system, and the green channel intensities of obtained colored images were extracted with photoshop software to quantify the luminescence. As a result, the quenching of the luminescence of the UCNPs is linearly correlated to the concentration of the exosomes (in the range of 1.0 × 104 ~ 1.0 × 108 particles/µL), enabling the detection and quantification of the exosomes. Such approach can reach a low detection limit of exosomes (1.1 × 103 particles/µL) and effectively reduce the background signal by using UCNPs as a luminescent material. This study provides an efficient and practical approach to the detection of exosomes, which should lead to point-of-care testing in clinical applications.


Assuntos
Técnicas Biossensoriais/métodos , Exossomos/química , Transferência Ressonante de Energia de Fluorescência/métodos , Nanopartículas Metálicas/química , Ouro/química , Humanos , Limite de Detecção , Luminescência , Nanotubos/química
19.
Inorg Chem ; 56(8): 4595-4601, 2017 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-28358498

RESUMO

Alkalides, as a representative kind of excess electron compounds, have been demonstrated to be potential nonlinear optical (NLO) materials with large static first hyperpolarizabilities (ß0). The possibility of utilizing coinage metal atoms as a novel excess electron source to design a series of alkalides, i.e., (M@36adz)M' (M = Cu, Ag, and Au; M' = Li, Na, and K), was examined by density functional theory calculations. The alkalide characteristics of these compounds are guaranteed by their HOMOs and VIE values as well as NBO analysis. In particular, all proposed alkalides exhibit considerable first hyperpolarizabilities (ß0) up to 61 590 au, indicating that they can be considered as novel NLO molecules of high performance. Moreover, a larger cage-complexant has been considered, and the resulting (Ag+@TriPip222)K- alkalide possesses a remarkably large ß0 value of 180 068 au. We hope that this work will provide a new recipe for designing excess electron compounds and, on the other hand, attract more research interest and efforts in exploring new, unconventional alkalides.

20.
Biosens Bioelectron ; 92: 8-15, 2017 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-28167415

RESUMO

Recently, many studies have shown the potential use of circulating exosomes as novel biomarkers for monitoring and predicting a number of complex diseases, including cancer. However, reliable and cost-effective detection of exosomes in routine clinical settings, still remain a difficult task, mainly due to the lack of adequately easy and fast assay platforms. Therefore, we demonstrate here the development of a visible and simple method for the detection of exosomes by integrating single-walled carbon nanotubes that being excellent water solubility (s-SWCNTs) and aptamer. Aptamers, specific to exosomes transmembrane protein CD63, are absorbed onto the surface of s-SWCNTs and improve the minic peroxidase activity of s-SWCNTs, which can efficiently catalyze H2O2-mediated oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) and lead to a change from colorless to blue in solution. However, after adding exosomes, the aptamers are bound with CD63, leaving from the surface of s-SWCNTs through conformational changes, which results the color of solution from deep to moderate, and this can be observed by the naked eye and monitored by UV-vis spectrometry. Under optimal conditions, the linear range of exosomes is estimated to be 1.84×106 to 2.21×107 particles/µL with a detection of limit (LOD) of 5.2×105 particles/µL. Consequently, a visible and simple approach detecting exosomes is successfully constructed. Moreover, this proposed colorimetric aptasensor can be universally applicable for the detection of other targets by simple change the aptamer.


Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Colorimetria/métodos , Exossomos/química , Nanotubos de Carbono/química , Tetraspanina 30/análise , Benzidinas/química , Catálise , Compostos Cromogênicos/química , Humanos , Peróxido de Hidrogênio/química , Células MCF-7 , Nanotubos de Carbono/ultraestrutura , Oxirredução , Peroxidase/química
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