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1.
Anal Chem ; 2023 Jan 12.
Artigo em Inglês | MEDLINE | ID: mdl-36633481

RESUMO

Owing to the excellent structural rigidity and programmable reaction sites, DNA nanostructures are more and more widely used, but they are limited by high cost, strict sequence requirements, and time-consuming preparation. Herein, a general signal amplifier based on a micelle-supported entropy-driven circuit (MEDC) was designed and prepared for sensitive quantification of biomarkers. By modifying a hydrophobic cholesterol molecule onto a hydrophilic DNA strand, the amphiphilic DNA strand was first prepared and then self-assembled into DNA micelles (DMs) driven by hydrophobic effects. The as-developed DM showed unique advantages of sequence-independence, easy preparation, and low cost. Subsequently, amplifier units DMF and DMTD were successfully fabricated by connecting fuel strands and three-strand duplexes (TDs) to DMs, respectively. Finally, the MEDC was triggered by microRNA-155 (miR-155), which herein acted as a model analyte, resulting in dynamic self-assembly of poly-DNA micelles (PDMs) and causing the recovery of cyanine 3 (Cy3) fluorescence as the DMTD dissociated. Benefiting from the "diffusion effect", the MEDC herein had a nearly 2.9-fold increase in sensitivity and a nearly 97-fold reduction in detection limit compared to conventional EDC. This amplifier exhibited excellent sensitivity of microRNAs, such as miR-155 detection in a dynamic range from 0.05 to 4 nM with a detection limit of 3.1 pM, and demonstrated outstanding selectivity with the distinguishing ability of a single-base mismatched sequence of microRNAs. Overall, the proposed strategy demonstrated that this sequence-independent DNA nanostructure improved the performance of traditional DNA probes and provided a versatile method for the development of DNA nanotechnology in biosensing.

2.
Anal Chem ; 95(19): 7603-7610, 2023 05 16.
Artigo em Inglês | MEDLINE | ID: mdl-37129512

RESUMO

System leakage critically confines the development of cascade DNA systems that need to be implemented in a strict order-by-order manner. In principle, ternary DNA reactants, composed of three single-strand DNA (ssDNA) with a strict equimolar ratio (1:1:1), have been indispensable for successfully cascading upstream entropy-driven DNA circuit (EDC) with downstream circuits, and system leakage will occur with any unbalance of the molar ratio. In this work, we proposed "splitting-reconstruction" and "protection-release" strategies on the potential downstream circuit initiator derived from upstream EDC to guide the construction of EDC-involved cascade systems independent of system leakage derived from unpurified reactants. Both the reconstructed and released downstream circuit initiators were in compliance with the principle of the cascade AND logic gate. Using these two strategies, two cascade systems─EDC2-4WJ-TMSDR and EDC3-HCR─were developed to carry out the designed order, which did not require that the ratio of 1:1:1 be maintained. Furthermore, the inherent property of the upstream EDC could transfer into the downstream circuit, endowing the developed cascade systems with a more powerful signal amplification ability for the sensitive detection of the corresponding initiator strand. These two strategies may provide new insights into the process of constructing EDC-like circuit-involved high-order DNA networks.


Assuntos
DNA de Cadeia Simples , DNA , DNA/genética , DNA de Cadeia Simples/genética , Entropia , Lógica
3.
Anal Chem ; 95(18): 7237-7243, 2023 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-37120835

RESUMO

DNA nanosheets (DNSs) have been utilized effectively as a fluorescence anisotropy (FA) amplifier for biosensing. But, their sensitivity needs to be further improved. Herein, CRISPR-Cas12a with strong trans-cleavage activity was utilized to enhance the FA amplification ability of DNSs for the sensitive detection of miRNA-155 (miR-155) as a proof-of-principle target. In this method, the hybrid of the recognition probe of miR-155 (T1) and a blocker sequence (T2) was immobilized on the surface of magnetic beads (MBs). In the presence of miR-155, T2 was released by a strand displacement reaction, which activated the trans-cleavage activity of CRISPR-Cas12a. The single-stranded DNA (ssDNA) probe modified with a carboxytetramethylrhodamine (TAMRA) fluorophore was cleaved in large quantities and could not bind to the handle chain on DNSs, inducing a low FA value. In contrast, in the absence of miR-155, T2 could not be released and the trans-cleavage activity of CRISPR-Cas12a could not be activated. The TAMRA-modified ssDNA probe remained intact and was complementary to the handle chain on the DNSs, and a high FA value was obtained. Thus, miR-155 was detected through the obviously decreased FA value with a low limit of detection (LOD) of 40 pM. Impressively, the sensitivity of this method was greatly improved about 322 times by CRISPR-Cas12a, confirming the amazing signal amplification ability of CRISPR-Cas12a. At the same time, the SARS-CoV-2 nucleocapsid protein was detected by the strategy successfully, indicating that this method was general. Moreover, this method has been applied in the analysis of miR-155 in human serum and the lysates of cells, which provides a new avenue for the sensitive determination of biomarkers in biochemical research and disease diagnosis.


Assuntos
Técnicas Biossensoriais , COVID-19 , MicroRNAs , Humanos , SARS-CoV-2 , DNA , DNA de Cadeia Simples , Sistemas CRISPR-Cas/genética
4.
Anal Chem ; 95(29): 10992-10998, 2023 07 25.
Artigo em Inglês | MEDLINE | ID: mdl-37436093

RESUMO

Challenges remained in precisely real-time monitoring of apoptotic molecular events at the subcellular level. Herein, we developed a new type of intelligent DNA biocomputing nanodevices (iDBNs) that responded to mitochondrial microRNA-21 (miR-21) and microRNA-10b (miR-10b) simultaneously which were produced during cell apoptosis. By hybridizing two hairpins (H1 and H2) onto DNA nanospheres (DNSs) that had been previously modified with mitochondria-targeted triphenylphosphine (TPP) motifs, iDBNs were assembled in which two localized catalytic hairpins self-assembly (CHA) reactions occurred upon the co-stimulation of mitochondrial miR-21 and miR-10b to perform AND logic operations, outputting fluorescence resonance energy transfer (FRET) signals for sensitive intracellular imaging during cell apoptosis. Owing to the spatial confinement effects of DNSs, it was discovered that iDBNs had a high efficiency and speed of logic operations by high local concentrations of H1 and H2, making the simultaneous real-time responses of mitochondrial miR-21 and miR-10b reliable and sensitive during cell apoptosis. These results demonstrated that iDBNs were simultaneously responsive to multiple biomarkers, which greatly improved the detection accuracy to identify the cell apoptosis, demonstrating that iDBNs are highly effective and reliable for the diagnosis of major disease and screening of anticancer drugs.


Assuntos
MicroRNAs , MicroRNAs/genética , DNA , Apoptose , Biomarcadores
5.
Anal Chem ; 94(39): 13440-13446, 2022 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-36130106

RESUMO

Plasmon-driven catalysis of metal nanostructures has garnered wide interest. Here, a photogenerated plasmonic hot-electron painting strategy was reported to form Au@Pt composite nanoparticles (Au@Pt NPs) with high catalytic reactivity without using reducing agents. Au nanoparticles, including Au nanospheres (Au NSs), Au nanorods (Au NRs), and Au nanobipyramids (Au NBPs), generated hot electrons under localized surface plasmon resonance (LSPR) excitation, which made the platinum precursor reduced as a consequence that Pt(0) atoms were painted on the surface of Au NPs to form an asymmetric Pt shell outside the plasmonic Au core. Compared with bare Au NPs, Au@Pt NPs exhibited significantly enhanced electrocatalytic activity toward reduction of H2O2 due to the bimetallic synergistic effect and great dispersion of Au@Pt NP-modified indium tin oxide (Au@Pt NPs/ITO). It exhibited a linear detection of H2O2 in a wide concentration range from 0.5 to 1000 µM with a low detection limit of 0.11 µM (S/N = 3). Therefore, the plasmonic hot-electron-painted Au@Pt NPs represent a novel and simple method for the design of advanced noble asymmetric metal nanomaterials.


Assuntos
Ouro , Nanopartículas Metálicas , Elétrons , Ouro/química , Peróxido de Hidrogênio/química , Nanopartículas Metálicas/química , Platina/química , Substâncias Redutoras
6.
Chemistry ; 28(54): e202201437, 2022 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-35735906

RESUMO

Metal-organic frameworks (MOFs) with abundant active sites, a class of materials composed of metal nodes and organic ligands, is widely used for photocatalytic degradation of pollutants. However, the rapid recombination of photoinduced carriers of MOFs limits its photocatalytic degradation performance. Herein, Ti3 C2 Tx nanosheets-based NH2 -MIL-101(Fe) hybrids with Schottky-heterojunctions were fabricated by in situ hydrothermal assembly for improved photocatalytic activity. The photodegradation efficiencies of the NH2 -MIL-101(Fe)/Ti3 C2 Tx (N-M/T) hybrids for phenol and chlorophenol were 96.36 % and 99.83 % within 60 minutes, respectively. The N-M/T Schottky-heterojunction duly transferred electrons to the Ti3 C2 Tx nanosheets surface via built-in electric fields, effectively suppressing the recombination of photogenerated carriers, thereby improving the photocatalytic performance of NH2 -MIL-101(Fe). Moreover, the Fe-mixed-valence in the N-M/T led to improvement in the efficiency of the in situ generated photo-Fenton reactions, further enhancing the photocatalytic activity with more generated reactive oxygen species (ROS). The study proposes a highly effective removal of phenolic pollutants in wastewater.


Assuntos
Clorofenóis , Poluentes Ambientais , Estruturas Metalorgânicas , Ligantes , Estruturas Metalorgânicas/química , Fenóis , Espécies Reativas de Oxigênio , Titânio , Águas Residuárias
7.
Mikrochim Acta ; 189(5): 181, 2022 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-35394213

RESUMO

A catalyst-free co-reaction luminol-H2O2-K2S2O8 chemiluminescence (CL) system was developed, with long-life and high-intensity emission, and CL emission lasting for 6 h. A possible mechanism of persistent and intense emission in this CL system was discussed in the context of CL spectra, cyclic voltammetry, electron spin resonance (ESR), and the effects of radical scavengers on luminol-H2O2-K2S2O8 system. H2O2 and K2S2O8 co-reactants can promote each other to continuously generate corresponding radicals (OH•, 1O2, O2•-, SO4•-) that trigger the CL emission of luminol. H2O2 can also be constantly produced by the reaction of K2S2O8 and H2O to further extend the persistence of this CL system. CL emission can be quenched via ascorbic acid (AA), which can be generated through hydrolysis reaction of L-ascorbic acid 2-phosphate trisodium salt (AAP) and alkaline phosphatase (ALP). Next, a CL-based method was established for the detection of ALP with good linearity from 0.08 to 5 U·L-1 and a limit of detection of 0.049 U·L-1. The proposed method was used to detect ALP in human serum samples.


Assuntos
Fosfatase Alcalina/análise , Luminescência , Luminol , Humanos , Peróxido de Hidrogênio , Limite de Detecção , Medições Luminescentes/métodos
8.
Angew Chem Int Ed Engl ; 61(11): e202115561, 2022 03 07.
Artigo em Inglês | MEDLINE | ID: mdl-34989066

RESUMO

Sensitive sensing is critical when developing new calculation systems with weak input signals (ISs). In this work, a "weak-inputs-strong-outputs" strategy was proposed to guide the construction of sensitive logic nanodevices by coupling an input-induced reversible DNA computing platform with a hybridization chain reaction-based signal amplifier. By rational design of the sequence of computing elements (CEs) so as to avoid cross-talking between ISs and signal amplifier, the newly formed logic nanodevices have good sensitivity to the weak ISs even at low concentrations of CEs, and are able to perform YES, OR, NAND, NOR, INHIBIT, INHIBIT-OR and number classifier operation, showing that the DNA calculation proceeds in dilute solution medium that greatly improves the calculation proficiency of logic nanodevices without the confinement of the lithography process in nanotechnology.

9.
Anal Chem ; 93(7): 3411-3417, 2021 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-33565859

RESUMO

In this work, we propose a three-layer hierarchical hybridization chain reaction (3L hHCR) composed of 1stHCR, 2ndHCR, and 3rdHCR to achieve robust signal amplification efficiency and broaden the applied range of HCR-based systems. In principle, the execution of superior HCR generates the formation of the initiator (named as 2ndI or 3rdI) of the subordinate HCR that relies on the introduction of the target sequence (1stI). To avoid the high background signal of the 3L hHCR system, a strategy of "splitting reconstruction" was adopted. The initiator of the subordinate HCR was designed as two separate fragments (splitting) that are obtained together (reconstruction) for the motivation of the subordinate HCR after the completion of the superior HCR. The implementation of the entire 3L hHCR system generates significant fluorescence recovery that derives from the impediment of Förster resonance energy transfer between fluorophore and quencher; thus, ultrasensitive detection of 1stI in the range of 50 pM to 10 nM can be achieved. Surprisingly, when the concentration of 1stI is lower than 1 nM, the 3L hHCR shows excellent ability to discriminate against various concentrations of 1stI, which is better than that of the 2L hHCR I system. Due to the hierarchical self-assembly mechanism, the 3L hHCR can also be reliably operated as a cascade AND logic gate with a high specificity and molecular keypad lock with a prompt error-reporting function. Furthermore, the 3L hHCR-based molecular keypad lock also shows potential application in the accurate diagnosis of cancer. The 3 L hHCR shows visionary prospects in biosensing and the fabrication of advanced biocomputing networks.

10.
Anal Chem ; 93(7): 3526-3534, 2021 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-33562958

RESUMO

Precise drug delivery holds great promise in cancer treatment but still faces challenges in controllable drug release in tumor cells specifically. Herein, a nucleolin-targeted and telomerase-responsive DNA nanotube for drug release was developed. First, a DNA nanosheet with four capture strands on its surface was prepared, which could bind and load ricin A chain (RTA). The RTA-loaded nanosheet was further converted into a DNA nanotube with a high Förster resonance energy transfer (FRET) efficiency in the presence of a Cy3-modified DNA fastener by hybridizing with the Cy5-modified DNA and another DNA-containing telomerase primer sequence along the long sides. Moreover, the aptamer of nucleolin was assembled on the DNA nanotube by combining with the hybrid chain at the terminal. The aptamer-functionalized and RTA-loaded DNA nanotube displayed enhanced tumor permeability and precise drug release in response to the telomerase in tumor cells, following the change of the FRET signal and RTA-induced cell death. Moreover, the DNA nanotube was applied successfully in vivo, and there was an obvious inhibition of tumor growth on xenograft-bearing mice following systemic administration, indicating that the constructed DNA nanotube represents a promising platform for precise RTA delivery in target cancer therapy.


Assuntos
Nanotubos , Neoplasias , Telomerase , Animais , DNA , Transferência Ressonante de Energia de Fluorescência , Camundongos , Neoplasias/tratamento farmacológico , Fosfoproteínas , Proteínas de Ligação a RNA , Telomerase/genética , Telomerase/metabolismo , Nucleolina
11.
Anal Chem ; 92(17): 11565-11572, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-32786463

RESUMO

Fluorescent labeled single-stranded DNA (ssDNA) molecules physisorbed on graphene oxide (GO) have been extensively explored as a useful sensing platform. However, this approach faces challenges when applied to complex biological samples due to heavy nonspecific desorption of nontarget molecules from GO. To overcome this problem, we introduced a capture DNA (cDNA) fragment with a poly adenine (poly-A) extension into the physisorption system that greatly reduces nonspecific desorption and false positive signal due to strong binding between poly-A and GO. Fluorescence from the dye can be effectively quenched by BHQ, which thus provides a second guarantee of anti-interference to avoid possible nonspecific poly-A DNA displacement. As a proof of concept, we have successfully developed a novel DNA-adsorbing GO nanocomplex probe (DNA-GO nanocomplex probe). This probe has a high anti-interference capability and low background due to the presence of both GO and black hole quencher (BHQ) as a dual-quencher that reduces the background in live cell imaging due to resonance energy transfer (RET). We then employed the DNA-GO nanocomplex probe for simultaneous detection of miR-630 and miR-21 and also for simultaneous in situ dynamic monitoring of intracellular miR-630 and miR-21 in apoptotic cells. We discovered that miR-630 expression was up-regulated during the first 120 min. This simple but powerful protocol has great potential in precise detection and imaging of various substances in complex biological samples with improved accuracy.


Assuntos
Sondas de DNA/química , DNA de Cadeia Simples/química , Grafite/química , MicroRNAs/análise , Nanoestruturas/química , Adsorção , Apoptose , Técnicas Biossensoriais , Transferência Ressonante de Energia de Fluorescência , Corantes Fluorescentes/química , Humanos , Imagem Óptica , Propriedades de Superfície
12.
Angew Chem Int Ed Engl ; 59(8): 3300-3306, 2020 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-31825124

RESUMO

Synthesizing 2D metal-organic frameworks (2D MOFs) in high yields and rational tailoring of the properties in a predictable manner for specific applications is extremely challenging. Now, a series of porphyrin-based 2D lanthanide MOFs (Ln-TCPP, Ln=Ce, Sm, Eu, Tb, Yb, TCPP=tetrakis(4-carboxyphenyl) porphyrin) with different thickness were successfully prepared in a household microwave oven. The as-prepared 2D Ln-TCPP nanosheets showed thickness-dependent photocatalytic performances towards photooxidation of 1,5-dihydroxynaphthalene (1,5-DHN) to synthesize juglone. Particularly, the Yb-TCPP displayed outstanding photodynamic activity to generate O2 - and 1 O2 . This work not only provides fundamental insights into structure designing and property tailoring of 2D MOFs nanosheets, but also pave a new way to improve the photocatalytic performance.

13.
Anal Chem ; 91(17): 11185-11191, 2019 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-31412694

RESUMO

Traditional fingerprints are usually obtained by pressing an inked finger on a paper. The inks would contaminate fingers and more importantly, these fingerprints are visible and able to be photocopied. In order to develop a smart membrane for fingerprint recording and document security, microrod assemblies of carbon quantum dots (CQDs)-Eu (III) are embedded in a electrospun nanofibrous (NFs) membrane which has strong red emission under UV irradiation owing to aggregation induced Dexter energy transfer from CQDs to Eu (III) ions. A clear blue emission fingerprint could be recorded on the membrane after a finger touch because the phosphate (Pi) secreted through sweat glands blocks the solid-state Dexter energy transfer, recovering the UV-irradiated blue emissions of CQDs. The Pi-based fingerprint on the membrane, which is invisible under daylight and could not be photocopied, greatly improves the security of the fingerprint and, furthermore, has the capability to identify the people who touched the secret document through the fingerprint analysis, showing that the intelligent NFs membrane can be applied for both fingerprint security and document counterspy.


Assuntos
Carbono/química , Európio/química , Tinta , Nanofibras/química , Papel , Pontos Quânticos/química , Transferência de Energia
14.
Mikrochim Acta ; 186(3): 168, 2019 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-30739202

RESUMO

A bimetallic organic gel was prepared by mixing the bridging ligand 2,4,6-tri(4-carboxyphenyl)-1,3,5-triazine with Cu(II) and Co(II) ions at room temperature. The resulting metal-organic gel (MOG) shows enhanced peroxidase-like activity, most likely due to the synergetic redox cycling between Co(III)/Co(II) and Cu(II)/Cu(I) pairs. These accelerate interfacial electron transfer and generation of hydroxy radicals. The MOG can catalyze the reaction of H2O2 with terephthalic acid (TPA), producing a blue fluorescence product with the maximum excitation/emission at 315/446 nm. The enzyme mimic was used to design a fluorometric method for H2O2 that has a 81 nM detection limit. H2O2 is also formed by glucose oxidase-assisted oxidation of glucose by oxygen, and an assay for glucose was worked out based on the above method. It has a 0.33 µM detection limit. This study may open up a new avenue to design and synthesize nanomaterial-based biomimetic catalysts with multiple metal synergistically enhanced catalytic activity for potential applications in biocatalysis, bioassays and nano-biomedicine. Graphical abstracts Schematic presentation of the synergic catalytic effect of Cu(II)/Co(II) bimetallic organic gel promoted by the redox cycle between Co(III)/Co(II) and Cu(II)/Cu(I) pairs. The bimetallic organic gel can catalyze the reaction of H2O2 with terephthalic acid, thereby producing a blue-fluorescent product.

15.
Analyst ; 143(23): 5764-5770, 2018 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-30334036

RESUMO

Ricin is an extremely potent ribosome-inactivating protein and serves as a likely food biocontaminant or biological weapon. Thus, simple, sensitive and accurate analytical assays capable of detecting ricin are urgently needed to be established. Herein, we present a novel method for ricin B-chain (RTB) detection by using two materials: (a) a highly efficient hybrid probe that was formed by linking a glucose oxidase (GOD)-encapsulated liposome (GOD-L) to magnetic beads (MBs) through hybridization between an aptamer and a blocker and (b) a new low-background g-C3N4-MnO2 sandwich nanocomposite that exhibits fluorescence resonance energy transfer (FRET) between the g-C3N4 nanosheet and MnO2. In the presence of RTB, the strong binding between RTB and the aptamer can release the blocker-linked liposome from the surface of the MBs. After magnetic separation, the decomposed liposome can release GOD to catalyze the oxidation of glucose, generating a certain amount of H2O2. Then, H2O2 can reduce MnO2 of the g-C3N4-MnO2 nanocomposite to Mn2+, which leads to the elimination of FRET. Thus, the fluorescence of the g-C3N4 nanosheet will be turned on. Because of the excellent signal amplification ability of liposome and the characteristic highly sensitive response of the g-C3N4-MnO2 nanocomposite toward H2O2, RTB could be detected sensitively based on the significantly enhanced fluorescent intensity. The linear range of detection was from 0.25 µg mL-1 to 50 µg mL-1 and the limit of detection (LOD) was 190 ng mL-1. Moreover, the proposed assay was successfully applied in the detection of the entire ricin toxin content in a castor seed.


Assuntos
Compostos de Manganês/química , Nanocompostos/química , Óxidos/química , Ricina/análise , Aptâmeros de Nucleotídeos/química , Ricinus communis/química , Fluorescência , Transferência Ressonante de Energia de Fluorescência/métodos , Contaminação de Alimentos/análise , Glucose Oxidase/química , Peróxido de Hidrogênio/química , Limite de Detecção , Lipossomos , Oligodesoxirribonucleotídeos/química , Sementes/química
16.
Anal Chem ; 89(17): 8766-8771, 2017 09 05.
Artigo em Inglês | MEDLINE | ID: mdl-28737379

RESUMO

Graphene oxide (GO) has been proven as an outstanding fluorescence anisotropy (FA) amplifier. Yet the traditional GO amplified FA assays lack high sensitivity because of the 1:1 binding ratio between target and dye-modified probe. Herein, we report a new target-catalyzed hairpin assembly (CHA), an enzyme-free DNA circuit, assisted GO amplified FA strategy for microRNA-21 (miRNA-21) detection. In the presence of miRNA-21, the CHA was initiated and plenty of H1-H2 duplexes were produced continuously. The obtained H1-H2 duplex could induce the formation of a H1-H2-probe DNA (pDNA) complex by the toehold-mediated strand exchange reaction, which led the dye-modified pDNA to leave away from the GO surface, resulting in a decreased FA of the system. By monitoring the decrease of FA, miRNA-21 could be detected in the range of 0-16 nM. The limit of detection (LOD, 3σ) was 47 pM, which was 194 times lower than that without CHA. In addition, the selectivity of this method has also been enhanced greatly as compared to that without CHA. Our method has great potential to be applied for detecting different types of targets and monitoring diverse molecular interactions by adapting the corresponding nucleotide sequence.


Assuntos
Técnicas Biossensoriais/métodos , DNA/química , Grafite/química , MicroRNAs/análise , Espectrometria de Fluorescência/métodos , Linhagem Celular Tumoral , DNA/genética , Polarização de Fluorescência , Células Endoteliais da Veia Umbilical Humana , Humanos , Sequências Repetidas Invertidas , Limite de Detecção , MicroRNAs/genética , Hibridização de Ácido Nucleico , Óxidos/química
17.
Anal Chem ; 88(4): 2250-7, 2016 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-26750592

RESUMO

During the past decade, aptasensors have largely been designed on the basis of the notion that ligand-modulated equilibration between aptamer conformations could be exploited for sensing. One implementation of this strategy has been to denature the aptamer with an antisense oligonucleotide, wait for dissociation of the antisense oligonucleotide, and stabilize the folded, signaling conformer with a ligand. However, there is a large kinetic barrier associated with releasing the oligonucleotide from the aptamer to again obtain an active, binding conformation. If the length of the antisense oligonucleotide is decreased to make dissociation from the aptamer more favorable, higher background signals are observed. To improve the general methodology for developing aptasensors, we have developed a novel and robust strategy for aptasensor design in which an oligonucleotide kinetically competes with the ligand for binding rather than having to be released from a stable duplex. While the oligonucleotide can induce conformational change, it initially chooses between the aptamer and a molecular beacon (MB), a process that does not require a lengthy pre-equilibration. Using an anti-ricin aptamer as a starting point, we developed a "competitive" aptasensor with a measured limit of detection (LOD) of 30 nM with an optical readout and as low as 3 nM for ricin toxin A-chain (RTA) detection on an electrochemical platform.


Assuntos
Aptâmeros de Nucleotídeos/análise , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais , Oligonucleotídeos Antissenso/química , Ligação Competitiva , Técnicas Eletroquímicas , Cinética , Ligantes , Limite de Detecção , Sondas Moleculares/análise , Sondas Moleculares/química , Ricina/análise , Ricina/química , Termodinâmica
18.
Analyst ; 140(1): 353-7, 2015 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-25375831

RESUMO

Fluorescence anisotropy (FA) has attracted considerable attention, but it has been rarely applied for the detection of small molecules and metal ions because they are too small to induce evident FA changes. Although some mass amplifying strategies have been developed, the recognition probes need to be covalently modified with the fluorescent dyes, which is complex and expensive. To overcome this limitation, a new simple, label-free and cost-effective method for the sensitive detection of potassium ion (K(+)), by using graphene oxide (GO) as FA enhancer, a G-rich single stranded DNA (ssDNA) as recognition probe and acridine orange (AO) as FA reporting fluorophore, was established in this paper. In the absence of K(+), both ssDNA and AO are adsorbed on the surface of GO, and the FA of AO is enhanced greatly because the rotation of AO is coupled with the entire formation. After the addition of K(+), the ssDNA self-associates into the G-quadruplex structure. Then, AO can bind with the formed G-quadruplex strongly, keeping away from the surface of GO, and the FA of AO decreases significantly because of the relatively small size of the complex of AO and G-quadruplex. Thus K(+) can be detected sensitively in the range of 10 µM-2 mM based on the evidently decreased FA. This method is a further improvement of the previous reported mass amplifying strategies because it does not require any covalent labelling of the recognition probe, and it can be potentially applied for detection of a variety of other targets.


Assuntos
Polarização de Fluorescência , Grafite/química , Óxidos/química , Potássio/análise , Íons/análise
19.
Anal Sci ; 40(3): 511-518, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38151695

RESUMO

Real-time monitoring of hypochlorous acid (HClO) in biological systems is of great significance for exploring and regulating its pathological functions because abnormal production of HClO is closely related with many diseases, such as atherosclerosis, rheumatoid arthritis, and cancer. Herein, we developed a reliable fluorescent Fe-doped carbon dots (Fe-CDs) for the sensitive and selective detection of biological HClO using ferrocenecarboxylic acid and m-phenylenediamine as precursors through a one-step solvothermal procedure. The Fe-CDs exhibited excellent a wide HClO detection range from 20 nmol/L to 1000 nmol/L with corresponding limits of detection at 7.8 nmol/L. The sensing mechanism is based on the chemical oxidation of the hydroxyl groups on the surface of Fe-CDs by HClO. In addition, Fe-CDs also displayed high photoluminescence yield, excitation-independence emission, as well as good biocompatibility, enabling the successful imaging of endogenous and exogenous HClO in HeLa cells. These results revealed that Fe-CDs holds great promise as a robust fluorescent probe for investigating HClO-mediated biological events.


Assuntos
Ácido Hipocloroso , Pontos Quânticos , Humanos , Células HeLa , Carbono , Corantes Fluorescentes
20.
Analyst ; 138(10): 3075-81, 2013 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-23586069

RESUMO

In this contribution, graphene oxide/gold nanoparticle (GO/AuNPs) hybrids were in situ fabricated through a green one-pot procedure by using tyrosine as an environment friendly and biocompatible reducing agent, which can be used as highly efficient surface enhanced Raman scattering (SERS) substrates with the enhancement factor at 3.8 × 10(3). The as-prepared GO/AuNPs hybrids have good biocompatibility, providing the prospect of applications for biomedicine determinations. In addition, taking the advantages of the electromagnetic and chemical enhancement mechanism and the high affinity of GO and AuNPs towards positive dyes, a sensitive, selective and label-free malachite green (MG) detection method was demonstrated. The SERS measurement showed that the minimum detection concentration of MG in water was as low as 2.5 µmol L(-1) with a linear response range from 2.5 to 100 µmol L(-1) (R(2) = 0.996). Moreover, this method can be applied to detect MG in a fishery water sample with satisfactory results.


Assuntos
Ouro/química , Grafite/química , Nanopartículas Metálicas/química , Óxidos/química , Corantes de Rosanilina/análise , Modelos Moleculares , Tamanho da Partícula , Análise Espectral Raman , Propriedades de Superfície , Água/química
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