Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 22
Filtrar
1.
Talanta ; 277: 126430, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38878510

RESUMO

In the present work, we developed an aptasensor to determine chloramphenicol (CAP) based on the dual signal output of photoelectrochemistry (PEC) and colorimetry. The Fe3+-doped porous tungsten trioxide was prepared by sol-gel method and coated on the ITO conductive glass to form ITO/p-W(Fe)O3. After assembling the captured DNA (cDNA) and the aptamer of CAP (apt) successively, the constructed ITO/p-W(Fe)O3-cDNA/apt aptasensor exhibited excellent photocurrent response under visible light irradiation in the presence of glucose, which provided the feasibility for PEC measurement with high sensitivity. In the presence of CAP, the apt left the ITO/p-W(Fe)O3 surface and AuNPs linked on the probe DNA would be assembled on it, which led to the decrease of photocurrent. Thanks to the oxidase-mimic catalytic performance of AuNPs and the recycling catalytic hydrolysis by exonuclease I, the measurement signal of the aptasensor could be amplified significantly, and the photocurrent decrease of the aptasensor was linearly related to the concentration of CAP in the range of 1.0 pM-10.0 nM and low detection limit was 0.36 pM. Meanwhile, the H2O2 produced from catalytic oxidation of glucose could oxidize TMB to blue oxTMB under HRP catalysis, which absorbance at 652 nm was linearly related to the concentration of CAP in the range of 5.0 pM-10.0 nM and low detection limit was 1.72 pM. Therefore, an aptasensor that determine CAP in real samples was successfully constructed with good precision of the relative standard deviation less than 5.7 % for PEC method and 7.3 % for colorimetric method, which can meet the analysis needs in different scenarios.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Cloranfenicol , Colorimetria , Técnicas Eletroquímicas , Ouro , Nanopartículas Metálicas , Cloranfenicol/análise , Cloranfenicol/química , Aptâmeros de Nucleotídeos/química , Colorimetria/métodos , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Ouro/química , Nanopartículas Metálicas/química , Limite de Detecção , Óxidos/química , Processos Fotoquímicos , Antibacterianos/análise , Antibacterianos/química , Tungstênio
2.
Anal Chim Acta ; 1293: 342284, 2024 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-38331552

RESUMO

In the present work, we developed a photoelectrochemical aptasensor to determine omethoate (OMT) based on the dual signal amplification of CeO2@MnO2 photocatalysis for glucose oxidation and exonuclease I-assisted cyclic catalytic hydrolysis. CeO2@MnO2 heterojunction material prepared by hydrothermal method was linked with captured DNA (cDNA) and then assembled on the ITO conductive glass to form ITO/CeO2@MnO2-cDNA, which exhibited significant photocurrent response and good photocatalytic performance for glucose oxidation under visible light irradiation, providing the feasibility for sensitive determining OMT. After binding with the aptamer of OMT (apt), the formation of rigid double stranded cDNA/apt kept CeO2@MnO2 away from ITO surface, which ensured a low photocurrent background for the constructed ITO/CeO2@MnO2-cDNA/apt aptasensor. In the presence of target OMT, the restoration of the cDNA hairpin structure and the exonuclease I-assisted cyclic catalytic hydrolysis led to the generation and amplification of measurement photocurrent signals, and allowed the aptasensor to have an ideal quantitative range of 0.01-10.0 nM and low detection limit of 0.0027 nM. Moreover, the aptasensor has been applied for selective determination of OMT in real samples with good precision of the relative standard deviation less than 6.2 % and good accuracy of the recoveries from 93 % to 108 %. What's more, the aptasensor can be used for other target determination only by replacing the captured DNA and corresponding aptamer.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Dimetoato/análogos & derivados , Glucose , DNA Complementar , Compostos de Manganês , Óxidos , DNA/química , Técnicas Biossensoriais/métodos , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Limite de Detecção
3.
Mikrochim Acta ; 190(12): 470, 2023 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-37971689

RESUMO

A portable colorimetric aptasensor was constructed based on the dual catalytic performance of CeO2 nanozyme to determine carbohydrate antigen 125 (CA125). Firstly, CeO2 nanozyme was synthesized by calcination and ultrasonically dispersed in a macroporous silica foam (MSF) to form CeO2@MSF. Then the aptamer of CA125 (apt) and complementary DNA (c-DNA) were successively assembled on the CeO2@MSF to construct a CeO2@MSF/apt/c-DNA colorimetric aptasensor, which exhibited excellent oxidase-mimic performance and phosphatase-mimic activity simultaneously. In the presence of CA125, the apt specifically binds to target CA125, and the single-strand c-DNA leaves the CeO2@MSF/apt surface, which is catalytically hydrolyzed by exonuclease I. The produced phosphate ions inhibit the phosphatase-mimic activity of CeO2 nanozyme. Thus, the absorbance at 652 nm of 3,3',5,5'-tetramethylbenzidine solution containing ascorbic acid-2-phosphate increases with the concentration of CA125. The response is linearly related to the logarithm of CA125 concentration from 1.0 to 10.0 U/mL under optimal experimental conditions. Based on this, the constructed colorimetric aptasensor has a high sensitivity, good selectivity, and high accuracy for CA125 determination in real human serum sample.


Assuntos
Colorimetria , Dióxido de Silício , Humanos , DNA de Cadeia Simples , Monoéster Fosfórico Hidrolases , Fosfatos
4.
Talanta ; 252: 123830, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36030738

RESUMO

Understanding the catalytic performance of nanozymes assembled in confined environment is an interesting topic. Herein, a three-dimensional nanozyme-catalytic nanoreactor was constructed by confining MOF-818 nanozyme in the pore of macroporous tungsten trioxide (p-WO3). The catalytic activity of MOF-818 assembled in-situ for the oxidation of 3,5-Di-tert-butylcatechol (3,5-DTBC) could be regulated by changing the pore size of p-WO3. Only when being confined in the pores of p-WO3 with an appropriate pore size, MOF-818 could exhibit high affinity towards 3,5-DTBC, and excellent catalytic activity for 3,5-DTBC oxidation, the catalytic rate constant kcat and Michaelis constant Km were determined to be 31.47 s-1 and 1.42 mM, respectively, and the maximum yield of 3,5-DTBC oxidation reached 95.2%. Furthermore, the as-constructed nanozyme-catalytic nanoreactor could be designed to construct a colorimetric aptasensor for selective determining cardiac troponin I based on the enzymatic inhibition effect and the exonuclease I-assisted target recycling signal amplification, which exhibited a good linear range of 50 fg mL-1 - 100 ng mL-1, low detection limit of 18 fg mL-1, and was applied for human serum analysis with RSD less than 5.2% and the recoveries ranged from 95% to 107%.


Assuntos
Técnicas Biossensoriais , Colorimetria , Humanos , Colorimetria/métodos , Troponina I , Técnicas Biossensoriais/métodos , Catálise , Oxirredução
5.
Talanta ; 233: 122577, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34215069

RESUMO

In the present work, a capture DNA (c-DNA) was immobilized on the TNA/g-C3N4 to develop a sensitive and selective TNA/g-C3N4/c-DNA photoelectrochemical aptasensor for determining thrombin. With the aid of the specific recognition of anti-thrombin aptamer towards thrombin, ingenious design of hairpin DNA, and exonuclease III-assisted recycling signal amplification, more nanoceria could be assembled on the TNA/g-C3N4/c-DNA to form TNA/g-C3N4/nanoceria in the presence of thrombin. Due to the oxidase-mimic catalytic efficiency of nanoceria and the oxygen consumption for glucose oxidation, the photoexcited electrons at the conduction band of g-C3N4 could be well transferred to that of TNA under visible light irradiation, resulting in the increase of the photocurrent of TNA/g-C3N4/nanoceria, and the increase value of photocurrent had a linear relationship with the concentration of thrombin under the optimal conditions. So, the constructed TNA/g-C3N4/c-DNA photoelectrochemical aptasensor exhibited a satisfactory quantitative range from 0.01 pM to 0.5 nM, low detection limit with 3.4 fM for thrombin determination, and was applied for the human serum analysis successfully with RSD of less than 4.8% and the recovery between 95% and 113%.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Cério , Técnicas Eletroquímicas , Exodesoxirribonucleases , Humanos , Limite de Detecção , Trombina
6.
Biosens Bioelectron ; 165: 112357, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32729490

RESUMO

A self-powered aptasensor for prostate specific antigen (PSA) based on a membraneless photoelectrochemical fuel cell (PEFC) with double photoelectrodes was constructed, in which, PSA-binding aptamer was electrostatically immobilized on the KOH-doped g-C3N4 modified TiO2 nanotube arrays (TNA/A-g-C3N4/aptamer), which was used as a photoanode, and Fe3+-doped CuBi2O4 modified indium doped tin oxide (ITO) substrate (ITO/CBFeO) was used as a photocathode. Under visible light irradiation, glucose was photocatalytically oxidized by A-g-C3N4 and generated H2O2 in situ, which was used as the electron acceptor for ITO/CBFeO photocathode, thus producing a high cell output response with a maximum output power of 133.5 µW cm-2 and an open circuit potential of 0.98 V. Due to the specific recognition of PSA by the aptamer and the output power decrease of the PEFC caused by the steric hindrance of the captured PSA on the TNA/A-g-C3N4, the PEFC could be used as a self-powered aptasensor for PSA with a quantitative range of 0.005-50 ng mL-1, a low detection limit of 1.3 pg mL-1 and good selectivity, and has been successfully applied for the analysis of real human serum samples with good precision of the relative standard deviation (RSD) less than 5.6% and good accuracy of the recoveries ranged from 91% to 108%.


Assuntos
Técnicas Biossensoriais , Calicreínas , Nanotubos , Antígeno Prostático Específico , Técnicas Eletroquímicas , Humanos , Peróxido de Hidrogênio , Masculino
7.
Biosens Bioelectron ; 135: 160-165, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-31009883

RESUMO

In the present work, a double photoelectrode system has been constructed for photoelectrochemically driven enzymatic bioconversion and determination of nifedipine. In which, the TiO2 nanotube arrays in-situ assembled with g-C3N4 (TNA/g-C3N4) was used as a photoanode, and a cytochrome P450 3A4 (CYP3A4) enzyme was immobilized in the porous ITO/CuO films to fabricate an ITO/CuO/CYP3A4 photocathode. The constructed double photoelectrode system had a significant photocurrent response compared to the single ITO/CuO/CYP3A4 or TNA/g-C3N4 under visible light irradiation. Under optimal conditions, the photocurrent of the double photoelectrode system had a high catalytic activity toward substrate nifedipine with kcat of 5.62 s-1 and catalytic efficiency with kcat/kmapp of 0.94 µM-1 s-1, and the bioconversion yield of nifedipine reached 22.1%. Furthermore, the constructed double photoelectrode system could be used to determine the nifedipine concentration with a high sensitivity of 2.46 µA µM-1 and a low detection limit of 0.015 µM. Therefore, the proposed double photoelectrode system can be used well for study enzyme biocatalysis for target bioconversion, and also has a potential application for toxicity analysis.


Assuntos
Nifedipino/análise , Vasodilatadores/análise , Biocatálise , Técnicas Biossensoriais/instrumentação , Citocromo P-450 CYP3A/química , Técnicas Eletroquímicas/instrumentação , Eletrodos , Enzimas Imobilizadas/química , Desenho de Equipamento , Luz , Nanotubos/química , Nanotubos/ultraestrutura , Titânio/química
8.
Talanta ; 196: 293-299, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30683366

RESUMO

A sensitive photoelectrochemical (PEC) biosensor for determination of alkaline phosphatase (ALP) activity was constructed based on a photo-excited electron transfer strategy. Immobilization of CdTe quantum dots (QDs) on TiO2 nanotube arrays (TNAs), addition of iron (III) and adenosine triphosphate (ATP) in turn can effectively adjust the photocurrent response of TNAs under visible light irradiation due to a photo-excited electron transfer process, and alkaline phosphatase (ALP) activity can be determined for its catalysis toward dephosphorylation of ATP. The preparation of CdTe QDs, construction of TNA/QD PEC biosensor and the mechanism of photo-excited electron transfer are investigated in the present work. Under the optimal experimental conditions, the TNA/QD PEC biosensor shows a low limits of detection (LODs) (0.05 U L-1) and limits of quantification detection (LOQs) (0.15 U L-1), wide linear range from 0.2 to 15 U L-1, and good selectivity towards ALP determination, which has been successfully applied for human serum analysis with good precision (RSD ≤ 5.4%) and high accuracy (recovery rate, 91-112%).


Assuntos
Fosfatase Alcalina/química , Técnicas Biossensoriais , Trifosfato de Adenosina/química , Compostos de Cádmio/química , Técnicas Eletroquímicas , Transporte de Elétrons , Ferro/química , Luz , Nanotubos/química , Nanotubos/efeitos da radiação , Pontos Quânticos/química , Telúrio/química , Titânio/química , Titânio/efeitos da radiação
9.
Biosens Bioelectron ; 110: 1-7, 2018 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-29573621

RESUMO

Inspired by the photoelectrochemical (PEC) properties of TiO2 nanotubes arrays (TNA) and their application as a super vessel for immobilizing biomolecules, we constructed an inhibition-effect PEC biosensor for determination of asulam based on the in-situ generation of CdS quantum dots (QDs) on TNA using an enzymatic reaction. Horseradish peroxidase (HRP) enzyme was covalently assembled on the inner-wall of TNAs, which exhibited good electrochemical and catalytic properties. In the mixture solution containing H2O2, CdY and S2O32-, HRP enzyme in TNAs catalyzed H2O2 reduce S2O32- to S2-. The generated S2- reacted with CdY to form CdS QDs in situ on the TNAs, improving the PEC performance of TNA under visible light irradiation. The photocurrent would decrease after addition of asulam due to its inhibitory effect towards HRP enzyme activity. Under the optimal experimental conditions, the constructed PEC TNA/HRP biosensor exhibited a satisfying linear range (0.02-2.0 ng mL-1), low limit of detection (4.1 pg mL-1) and good selectivity towards asulam determination, and has been successfully applied for the analysis of real environmental water samples with good accuracy of the recoveries ranged from 90% to 114%.


Assuntos
Técnicas Biossensoriais/métodos , Carbamatos/análise , Nanotubos/química , Pontos Quânticos/química , Titânio/química , Poluentes Químicos da Água/análise , Água Potável/análise , Técnicas Eletroquímicas/métodos , Monitoramento Ambiental/métodos , Água Doce/análise , Peroxidase do Rábano Silvestre/química , Peróxido de Hidrogênio/química , Luz , Limite de Detecção , Nanotubos/ultraestrutura , Pontos Quânticos/ultraestrutura
10.
Anal Chim Acta ; 1000: 265-272, 2018 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-29289319

RESUMO

In the present work, colloidal cerium oxide nanoparticles (nanoceria) and graphene quantum dots (GQDs) were firstly synthesized by sol-gel method and pyrolysis respectively, which all have a uniform nano-size and significant fluorescence emission. Due to the fluorescence emission spectrum of nanoceria overlapped the absorption spectrum of GQDs, fluorescence resonance energy transfer (FRET) between nanoceria and GQDs could occur effectively by the electrostatic interaction. Based on it, a sensitive ratiometric fluorescence aptasensor for the determination of ochratoxin A (OTA), a small molecular mycotoxin produced by Aspergillus and Penicillium strains, has been successfully constructed. In which, probe DNA1@nanoceria and DNA2@GQD were designed to complementary with OTA aptamer, both could adsorb each other, leading to the occur of FRET. After adding of OTA aptamer and then introducing of OTA, the FRET would be interrupted/recovered due to the specific affinity of OTA and its aptamer, the fluorescence recovery value would increase with the addition of OTA. Under the optimal experimental conditions (pH 7, mGQD/nanoceria 2, captamer 100 nM, incubation time 30 min), the constructed ratiometric fluorescence aptasensor exhibited a satisfying linear range (0.01-20 ng mL-1), low limit of detection (2.5 pg mL-1) and good selectivity towards OTA, and has been successfully applied for the analysis of real sample peanuts with good accuracy of the recoveries ranged from 90 to 110%.


Assuntos
Aptâmeros de Nucleotídeos/química , Transferência Ressonante de Energia de Fluorescência , Grafite/química , Nanopartículas/química , Ocratoxinas/análise , Pontos Quânticos
11.
Analyst ; 142(5): 691-699, 2017 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-28180217

RESUMO

Electrode-electrolyte interface (EEI) analysis is of great significance in the investigation of electrochemical (EC) processes. Current ex situ surface techniques provided rich information of the structural/chemical composition of the EEI. However, real-time and in situ monitoring of the EEI is in great demand for a better understanding of the dynamic changes of the redox systems. Mass spectrometry (MS) is a useful technique for mass-resolved analysis of various analytes with the capability of on-line measurement. In this review, we give a brief overview of the development of online EC-MS techniques. Recent advances of well-established EC-MS techniques, such as differential electrochemical mass spectrometry and EC-electrospray ionization mass spectrometry, and their applications are summarized. Specifically, some newly designed EC reactors coupled with state-of-the-art mass spectrometry for the detection of short-lived intermediate species are highlighted.

12.
Chem Commun (Camb) ; 52(49): 7703-6, 2016 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-27230023

RESUMO

A light-driven approach combined with a macroporous reactor for the enzymatic biocatalytic reaction has been developed by confining the enzyme/photosensitizer nanohybrids in a macroporous material, which exhibits high bio-conversion efficiency due to the fast diffusion and collision between the enzyme/photosensitizer nanohybrid and the substrate in the reactor.

13.
Anal Chim Acta ; 902: 107-114, 2016 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-26703259

RESUMO

Cystatin C (CysC) is a sensitive marker for the estimation of the glomerular filtration rate and the clinical diagnosis of different diseases. In this paper, CysC-specific nanobodies (Nbs) were isolated from a phage display nanobody library. A simple and sensitive photoelectrochemical immunosensor based on TiO2 nanotube arrays (TNAs) was proposed for the sensitive detection of CysC. The TiO2 nanotube arrays deposited by electrochemical anodization displayed a high and stable photocurrent response under irradiation. After coupling CysC-specific nanobody to TNA (Nb/TNA), the proposed immunosensor for CysC can be utilized for tracking the photocurrent change of Nb/TNA caused by immunoreactions between CysC and the immobilized CysC-specific Nb. This allowed for the determination of CysC with a calibration range from 0.72 pM to 7.19 nM. The variation of the photocurrent was in a linear relationship with the logarithm of the CysC concentration in the range of 0.72 pM-3.6 nM. The immunosensor had a correlation coefficient of 0.97 and a detection limit of 0.14 pM at a signal-to-noise ratio of 3. The proposed immunosensor showed satisfactory intra- and inter-assay accuracy, high selectivity and good stability. As a result, this proposed strategy would offer a novel and simple approach for the detection of immunoreactions, provide new insights in popularizing the diagnosis of CysC, and extend the application of TiO2 nanotubes.


Assuntos
Técnicas Biossensoriais , Cistatina C/sangue , Nanotubos , Anticorpos de Domínio Único , Titânio/química , Eletroquímica , Humanos , Limite de Detecção , Microscopia Eletrônica de Varredura , Fotoquímica , Ressonância de Plasmônio de Superfície , Temperatura
14.
Anal Bioanal Chem ; 407(20): 6117-26, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26105511

RESUMO

Novel chemiluminescent (CL) imaging microtiter plates with high-throughput, low-cost, and simple operation for detection of four biomarkers related to Down's syndrome screening were developed and evaluated. To enhance the sensitivity of CL immunosensing, soybean peroxidase (SBP) was used instead of horseradish peroxide (HRP) as a label enzyme. The microtiter plates were fabricated by simultaneously immobilizing four capture monoclonal antibodies, anti-inhibin-A, anti-unconjugated oestriol (anti-uE3), anti-alpha-fetoprotein (anti-AFP), and beta anti-HCG (anti-ß-HCG), on nitrocellulose (NC) membrane to form immunosensing microtiter wells. Under a sandwiched immunoassay, the CL signals on each sensing site of the microtiter plates were collected by a charge-coupled device (CCD), presenting an array-based chemiluminescence imaging method for detection of four target antigens in a well at the same time. The linear response to the analyte concentration ranged from 0.1 to 40 ng/mL for inhibin-A, 0.075 to 40 ng/mL for uE3, 0.2 to 400 ng/mL for AFP, and 0.4 to 220 ng/mL for ß-HCG. The proposed microtiter plates possessed high-throughput, good stability, and acceptable accuracy for detection of four antigens in clinical serum samples and demonstrated potential for practical applicability of the proposed method to Down's syndrome screening. Graphical Abstract Schematic evaluation of the microtiter plater for simultaneous detection of the four biomarkers.


Assuntos
Biomarcadores/sangue , Síndrome de Down/diagnóstico , Glycine max/enzimologia , Ensaios de Triagem em Larga Escala/instrumentação , Medições Luminescentes/instrumentação , Peroxidase/química , Anticorpos Imobilizados/química , Biomarcadores/análise , Gonadotropina Coriônica Humana Subunidade beta/análise , Gonadotropina Coriônica Humana Subunidade beta/sangue , Síndrome de Down/sangue , Desenho de Equipamento , Estriol/análise , Estriol/sangue , Feminino , Humanos , Imunoensaio/métodos , Inibinas/análise , Inibinas/sangue , Gravidez , Diagnóstico Pré-Natal/instrumentação , Sensibilidade e Especificidade , alfa-Fetoproteínas/análise
15.
Analyst ; 140(2): 567-73, 2015 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-25417796

RESUMO

MicroRNA (miRNA) is found to be up-regulated in many kinds of cancer and therefore is classified as an oncomiR. Herein, we design a multifunctional fluorescent nanoprobe (FSiNP-AS/MB) with the AS1411 aptamer and a molecular beacon (MB) co-immobilized on the surface of the fluorescent dye-doped silica nanoparticles (FSiNPs) for target-cell-specific delivery and intracellular miRNA imaging. The FSiNPs were prepared by a facile reverse microemulsion method from tetraethoxysilane and silane derivatized coumarin that was previously synthesized by click chemistry. The as-prepared FSiNPs possess uniform size distribution, good optical stability and biocompatibility. In addition, there is a remarkable affinity interaction between the AS1411 aptamer and the nucleolin protein on the cancer cell surface. Thus, a target-cell-specific delivery system by the FSiNP-AS/MB is proposed for effectively transferring a MB into the cancer cells to recognize the target miRNA. Using miRNA-21 in MCF-7 cells (a human breast cancer cell line) as a model, the proposed multifunctional nanosystems not only allow target-cell-specific delivery with the binding affinity of AS1411, but also can track simultaneously the transfected cells and detect intracellular miRNA in situ. The proposed multifunctional nanosystems are a promising platform for a highly sensitive luminescent nonviral vector in biomedical and clinical research.


Assuntos
MicroRNAs/análise , MicroRNAs/genética , Nanopartículas , Neoplasias/diagnóstico , Aptâmeros de Nucleotídeos/metabolismo , Linhagem Celular Tumoral , Cumarínicos/química , Diagnóstico por Imagem/métodos , Corantes Fluorescentes/química , Humanos , Células MCF-7 , Neoplasias/metabolismo , Oligodesoxirribonucleotídeos/metabolismo , Fosfoproteínas/metabolismo , Proteínas de Ligação a RNA/metabolismo , Silanos/química , Nucleolina
16.
Langmuir ; 30(39): 11833-40, 2014 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-25222611

RESUMO

Cytochrome P450 enzymes (cyt P450s) with an active center of iron protoheme are involved in most clinical drugs metabolism process. Herein, an electrochemical platform for the investigation of drug metabolism in vitro was constructed by immobilizing cytochrome P450 2D6 (CYP2D6) with cyt P450 reductase (CPR) on graphene modified glass carbon electrode. Direct and reversible electron transfer of the immobilized CYP2D6 with the direct electron transfer constant of 0.47 s(-1) and midpoint potential of -0.483 V was obtained. In the presence of substrate tramadol, the electrochemical-driven CYP2D6 mediated catalytic behavior toward the conversion of tramadol to o-demethyl-tramadol was confirmed. The Michaelis-Menten constant (Km(app)) and heterogeneous reaction rate constant during the metabolism of tramadol were calculated to be 23.85 µM and 1.96 cm s(-1), respectively. The inhibition effect of quinidine on CYP2D6 catalyze-cycle was also investigated. Furthermore, this system was applied to studying the metabolism of other drugs.


Assuntos
Citocromo P-450 CYP2D6/química , Citocromo P-450 CYP2D6/metabolismo , Grafite/química , Nanocompostos/química , Tramadol/química , Tramadol/metabolismo , Biocatálise/efeitos dos fármacos , Inibidores do Citocromo P-450 CYP2D6/farmacologia , Eletroquímica , Eletrodos , Transporte de Elétrons , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Isoenzimas/antagonistas & inibidores , Isoenzimas/química , Isoenzimas/metabolismo , Modelos Moleculares , Polietilenoimina/química , Conformação Proteica , Quinidina/farmacologia
17.
Chem Commun (Camb) ; 50(90): 13896-9, 2014 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-25264962

RESUMO

A graphene nano-cage with regulatable space for the assembly of a cytochrome P450 1A2-UDP-glucuronosyltransferase 1A10 bienzyme complex has been constructed via a click reaction, and successfully used to study drug sequential metabolism using an electrochemically-driven method.


Assuntos
Citocromo P-450 CYP1A2/metabolismo , Técnicas Eletroquímicas , Glucuronosiltransferase/metabolismo , Grafite/metabolismo , Nanoestruturas/química , Preparações Farmacêuticas/metabolismo , Citocromo P-450 CYP1A2/química , Glucuronosiltransferase/química , Grafite/química , Humanos , Preparações Farmacêuticas/química
18.
Anal Chem ; 86(15): 8003-9, 2014 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-25014006

RESUMO

Understanding the enzymatic reaction kinetics that occur within a confined space or interface is a significant challenge. Herein, a nanotube array enzymatic reactor (CYP2C9/Au/TNA) was constructed by electrostatically adsorbing enzyme on the inner wall of TiO2 nanotube arrays (TNAs). TNAs with different dimensions could be fabricated by the anodization of titanium foil through varying the anodization potential or time. The electrical conductivity of TNAs was improved by electrodepositing Au nanoparticles on the inner wall of TNAs. The cytochrome P450 2C9 enzyme (CYP2C9) was confined inside TNAs as a model. The enzymatic activity of CYP2C9 and tolbutamide metabolic yield could be effectively regulated by changing the nanotube diameter and length of TNAs. The enzymatic rate constant k(cat) and apparent Michaelis constant K(m)(app) were determined to be 9.89 s(-1) and 4.8 µM at the tube inner diameter of about 64 nm and length of 1.08 µm. The highest metabolic yield of tolbutamide reached 14.6%. Furthermore, the designed nanotube array enzymatic reactor could be also used in situ to monitor the tolbutamide concentration with sensitivity of 28.8 µA mM(-1) and detection limit of 0.52 µM. Therefore, the proposed nanotube array enzymatic reactor was a good vessel for studying enzyme biocatalysis and drug metabolism, and has potential applications including efficient biosensors and bioreactors for chemical synthesis.


Assuntos
Citocromo P-450 CYP2C9/metabolismo , Hipoglicemiantes/farmacocinética , Nanotubos , Titânio/química , Tolbutamida/farmacocinética , Técnicas Eletroquímicas , Microscopia Eletrônica de Varredura
19.
Anal Chem ; 86(7): 3602-9, 2014 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-24576151

RESUMO

MicroRNAs (miRNAs) has been identified as diagnostic and prognostic biomarkers and predictors of drug response for many diseases, including a broad range of cancers, heart disease, and neurological diseases. The noninvasive theranostics system for miRNAs is very important for diagnosis and therapy of the cellular disease. Herein, a target-cell-specific theranostics nanoprobe for target-cell-specific delivery, cancer cells and intracellular miRNA-21 imaging, and cancer cell growth inhibition was proposed. The nanoprobe (FS-AS/MB) was prepared by simultaneously coupling of the AS1411 aptamer and miRNA-21 molecular beacon (miR-21-MB) onto the surface of Ru(bpy)3²âº-encapsulated silica (FS) nanoparticles. The FS nanoparticles synthesized by a facile reverse microemulsion method showed nearly monodisperse spherical shape with a smooth surface, good colloidal stability, a fluorescence quantum yield of ~21%, and low cytotoxicity. The antibiofouling polymer PEG grafted onto a silica shell reduced nonspecific uptake of cells. The ability of FS-AS/MB for target-specific cells delivery, simultaneous cancer cells, intracellular miRNA-21 imaging, and inhibition of miRNA-21 function and suppression of cell growth in vitro, were also demonstrated. The results of the present study suggested that the proposed nanoprobes would be a promising theranostics for different cancers by imaging and inhibiting other intracellular genes.


Assuntos
Sondas Moleculares , Nanopartículas , Neoplasias/patologia , Dióxido de Silício/química , Sequência de Bases , Primers do DNA , Fluorescência , Humanos , Células MCF-7
20.
Analyst ; 138(5): 1483-9, 2013 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-23330150

RESUMO

A rapid sandwiched immunoassay of microcystin-LR (MC-LR) in water is proposed with flow injection chemiluminescence detection. The magnetic beads (MBs) were first modified with polyethyleneimine (PEI) by acylamide bond between the carboxyl group on the surface of MBs and the primary amine group in PEI, followed by immobilizing of anti-MC-LR (Ab1) onto PEI with glutaraldehyde as linkage. The resulting Ab1 modified MBs captured the target MC-LR in water, reacted with the horseradish peroxidase and anti-MC-LR co-immobilized silica nanoparticles, and were detected with flow injection chemiluminescence. When using PEI/MBs as the carrier of anti-MC-LR, the CL signal was greatly enhanced up to 9-fold compared to that using MBs without PEI modification. The CL signal was further amplified 13-fold when Si/Ab2 was used as the signal probe. Under the optimal conditions, the present immunoassay exhibited a wide quantitative range from 0.02 to 200 µg L(-1) with a detection limit of 0.006 µg L(-1), which was much lower than the WHO provisional guideline limit of 1.0 µg L(-1) for MC-LR in drinking water. The relative standard deviation was 4.8% and the recoveries for the spiked samples ranged from 84% to 115%, which indicated acceptable precision and accuracy for MC-LR. The present method is easier to perform and less time-consuming (the entire analysis process lasted about 40 minutes) and has been applied to the detection of MC-LR in different water samples successfully.


Assuntos
Imunoensaio/métodos , Medições Luminescentes/métodos , Imãs/química , Microcistinas/análise , Polietilenoimina/química , Dióxido de Silício/química , Anticorpos Imobilizados/química , Água Potável/microbiologia , Enzimas Imobilizadas/química , Análise de Injeção de Fluxo/métodos , Peroxidase do Rábano Silvestre/química , Lagos/microbiologia , Limite de Detecção , Toxinas Marinhas , Sensibilidade e Especificidade
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA