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1.
J Chem Phys ; 160(6)2024 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-38341797

RESUMO

Diffusion of electrons over distances on the order of 100 µm has been observed in crystals of a small tetraheme cytochrome (STC) from Shewanella oneidensis [J. Huang et al. J. Am. Chem. Soc. 142, 10459-10467 (2020)]. Electron transfer between hemes in adjacent subunits of the crystal is slower and more strongly dependent on temperature than had been expected based on semiclassical electron-transfer theory. We here explore explanations for these findings by molecular-dynamics simulations of crystalline and monomeric STC. New procedures are developed for including time-dependent quantum mechanical energy differences in the gap between the energies of the reactant and product states and for evaluating fluctuations of the electronic-interaction matrix element that couples the two hemes. Rate constants for electron transfer are calculated from the time- and temperature-dependent energy gaps, coupling factors, and Franck-Condon-weighted densities of states using an expression with no freely adjustable parameters. Back reactions are considered, as are the effects of various protonation states of the carboxyl groups on the heme side chains. Interactions with water are found to dominate the fluctuations of the energy gap between the reactant and product states. The calculated rate constant for electron transfer from heme IV to heme Ib in a neighboring subunit at 300 K agrees well with the measured value. However, the calculated activation energy of the reaction in the crystal is considerably smaller than observed. We suggest two possible explanations for this discrepancy. The calculated rate constant for transfer from heme I to II within the same subunit of the crystal is about one-third that for monomeric STC in solution.


Assuntos
Citocromos , Elétrons , Transporte de Elétrons , Citocromos/química , Citocromos/metabolismo , Simulação de Dinâmica Molecular , Heme/química , Oxirredução
2.
Small ; 19(52): e2304013, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37653599

RESUMO

The ability to redirect electron transport to new reactions in living systems opens possibilities to store energy, generate new products, or probe physiological processes. Recent work by Huang et al. showed that 3D crystals of small tetraheme cytochromes (STC) can transport electrons over nanoscopic to mesoscopic distances by an electron hopping mechanism, making them promising materials for nanowires. However, fluctuations at room temperature may distort the nanostructure, hindering efficient electron transport. Classical molecular dynamics simulations of these fluctuations at the nano- and mesoscopic scales allowed us to develop a graph network representation to estimate maximum electron flow that can be driven through STC wires. In longer nanowires, transient structural fluctuations at protein-protein interfaces tended to obstruct efficient electron transfer, but these blockages are ameliorated in thicker crystals where alternative electron transfer pathways become more efficient. The model implies that more flexible proteinprotein interfaces limit the required minimum diameter to carry currents commensurate with conventional electronics.


Assuntos
Nanofios , Transporte de Elétrons , Citocromos/química , Citocromos/metabolismo , Simulação de Dinâmica Molecular , Proteínas/metabolismo
3.
Sensors (Basel) ; 23(3)2023 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-36772386

RESUMO

In view of the great threat of chloramphenicol (CAP) to human health and the fact that a few producers have illegally used CAP in the food production process to seek economic benefits in disregard of laws and regulations and consumer health, we urgently need a detection method with convenient operation, rapid response, and high sensitivity capabilities to detect CAP in food to ensure people's health. Herein, a molecularly imprinted polymer (MIP) electrochemical sensor based on a dual-signal strategy was designed for the highly sensitive analysis of CAP in milk. The NiFe Prussian blue analog (NiFe-PBA) and SnS2 nanoflowers were modified successively on the electrode surface to obtain dual signals from [Fe(CN)6]3-/4- at 0.2 V and NiFe-PBA at 0.5 V. SiO2-COOH@MIPs that could specifically recognize CAP were synthesized via thermal polymerization using carboxylated silica microspheres (SiO2-COOH) as carriers. When the CAP was adsorbed by SiO2-COOH@MIPs, the above two oxidation peak currents decreased at the same time, allowing the double-signal analysis. The SiO2-COOH@MIPs/SnS2/NiFe-PBA/GCE sensor used for determining CAP was successfully prepared. The sensor utilized the interactions of various nanomaterials to achieve high-sensitivity dual-signal detection, which had certain innovative significance. At the same time, the MIPs were synthesized using a surface molecular imprinting technology, which could omit the time of polymerization and elution and met the requirements for rapid detection. After optimizing the experimental conditions, the detection range of the sensor was 10-8 g/L-10-2 g/L and the limit of detection reached 3.3 × 10-9 g/L (S/N = 3). The sensor had satisfactory specificity, reproducibility, and stability, and was successfully applied to the detection of real milk samples.


Assuntos
Impressão Molecular , Dióxido de Silício , Humanos , Animais , Dióxido de Silício/química , Polímeros/química , Cloranfenicol , Leite , Reprodutibilidade dos Testes , Impressão Molecular/métodos , Técnicas Eletroquímicas/métodos , Eletrodos , Limite de Detecção
4.
Analyst ; 147(4): 712-721, 2022 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-35080213

RESUMO

In this work, a new type of Au-tetrahedral DNA nanostructure (Au-TDN) was originally proposed and successfully applied in an electrochemiluminescence aptasensor to detect organophosphorus pesticides (Ops). The aptamers modified with -SH could be covalently bonded with gold nanoparticles (AuNPs) to form a tetrahedron structure, and there were independent probes at each vertex of the tetrahedron, which could increase the probability of specific binding with Ops. The originally designed structure could not only maintain a stable tetrahedral configuration, but also combined with the target to improve the sensitivity of the sensor. Meanwhile, silver nanoparticles (AgNPs) could catalyze the chemical reaction between luminol and H2O2 to generate a variety of intermediates called reactive oxygen species (ROS) for signal enhancement. Factors that had important influences on the aptasensor, such as the concentration of Au-TDN, the incubation time, and the pH value of the buffer, were optimized in this trial. According to the final results, the limit of detection (LOD) of 3 pg mL-1 (S/N = 3) for methyl parathion, the LOD of 0.3 pg mL-1 (S/N = 3) for parathion and the LOD of 0.03 pg mL-1 (S/N = 3) for phoxim were obtained, respectively. Moreover, the novel tetrahedral structure could be replaced by different types of aptamers to expand its application range and lay a foundation for the development of portable rapid detection devices for pesticide residues.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Nanoestruturas , Praguicidas , DNA , Técnicas Eletroquímicas , Ouro , Peróxido de Hidrogênio , Limite de Detecção , Luminol , Compostos Organofosforados , Prata
5.
Sensors (Basel) ; 22(15)2022 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-35957269

RESUMO

In this work, we reported a rapid and sensitive fluorescence assay in homogenous solution for detecting organophosphorus pesticides by using tetramethylrhodamine (TAMRA)-labeled aptamer and its complementary DNA (cDNA) with extended guanine (G) bases. The hybridization of cDNA and aptamer drew TAMRA close to repeated G bases, then the fluorescence of TAMRA was quenched by G bases due to the photoinduced electron transfer (PET). Upon introducing the pesticide target, the aptamer bound to pesticide instead of cDNA because of the competition between pesticide and cDNA. Thus, the TAMRA departed from G bases, resulting in fluorescence recovery of TAMRA. Under optimal conditions, the limits of detection for phorate, profenofos, isocarbophos, and omethoate were 0.333, 0.167, 0.267, and 0.333 µg/L, respectively. The method was also used in the analysis of profenofos in vegetables. Our fluorescence design was simple, rapid, and highly sensitive, which provided a means for monitoring the safety of agricultural products.


Assuntos
Aptâmeros de Nucleotídeos , Praguicidas , Aptâmeros de Nucleotídeos/genética , DNA Complementar , Fluorescência , Compostos Organofosforados/análise , Praguicidas/análise
6.
Nano Lett ; 20(1): 208-217, 2020 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-31747755

RESUMO

Enzymes of natural biochemical pathways are routinely subcellularly organized in space and time in order to improve pathway efficacy and control. Designer scaffolding platforms are under development to confer similar benefits upon engineered pathways. Herein, we evaluate bacterial microcompartment shell (pfam0936-domain) proteins as modules for constructing well-defined nanometer scale scaffolds in vivo. We use a suite of visualization techniques to evaluate scaffold assembly and dynamics. We demonstrate recruitment of target cargo molecules onto assembled scaffolds by appending reciprocally interacting adaptor domains. These interactions can be refined by fine-tuning the scaffold expression level. Real-time observation of this system reveals a nucleation-limited step where multiple scaffolds initially form within a cell. Over time, nucleated scaffolds reorganize into a single intracellular assembly, likely due to interscaffold competition for protein subunits. Our results suggest design considerations for using self-assembling proteins as building blocks to construct nanoscaffolds, while also providing a platform to visualize scaffold-cargo dynamics in vivo.


Assuntos
Bactérias/química , Nanoestruturas/química , Bactérias/ultraestrutura , Nanoestruturas/ultraestrutura
7.
J Am Chem Soc ; 142(23): 10459-10467, 2020 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-32406683

RESUMO

Rapid and directed electron transfer (ET) is essential for biological processes. While the rates of ET over 1-2 nm in proteins can largely be described by simplified nonadiabatic theory, it is not known how these processes scale to microscopic distances. We generated crystalline lattices of Small Tetraheme Cytochromes (STC) forming well-defined, three-dimensional networks of closely spaced redox centers that appear to be nearly ideal for multistep ET. Electrons were injected into specific locations in the STC crystals by direct photoreduction, and their redistribution was monitored by imaging. The results demonstrate ET over mesoscopic to microscopic (∼100 µm) distances through sequential hopping in a biologically based heme network. We estimate that a hypothetical "nanowire" composed of crystalline STC with a cross-section of about 100 cytochromes could support the anaerobic respiration of a Shewanella cell. The crystalline lattice insulates mobile electrons from oxidation by O2, as compared to those in cytochromes in solution, potentially allowing for efficient delivery of current without production of reactive oxygen species. The platform allows direct tests of whether the assumptions based on short-range ET hold for sequential ET over mesoscopic distances. We estimate that the interprotein ET across 6 Å between hemes in adjacent proteins was about 105 s-1, about 100-fold slower than expectations based on simplified theory. More detailed analyses implied that additional factors, possibly contributed by the crystal lattice, may strongly impact mesoscale ET mainly by increasing the reorganizational energy of interprotein ET, which suggests design strategies for engineering improved nanowires suitable for future bioelectronic materials.


Assuntos
Citocromos/metabolismo , Cristalografia por Raios X , Citocromos/química , Transporte de Elétrons , Modelos Moleculares , Shewanella/química , Shewanella/citologia
8.
Appl Environ Microbiol ; 83(9)2017 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-28235875

RESUMO

Cyanobacteria are emerging as alternative crop species for the production of fuels, chemicals, and biomass. Yet, the success of these microbes depends on the development of cost-effective technologies that permit scaled cultivation and cell harvesting. Here, we investigate the feasibility of engineering cell morphology to improve biomass recovery and decrease energetic costs associated with lysing cyanobacterial cells. Specifically, we modify the levels of Min system proteins in Synechococcus elongatus PCC 7942. The Min system has established functions in controlling cell division by regulating the assembly of FtsZ, a tubulin-like protein required for defining the bacterial division plane. We show that altering the expression of two FtsZ-regulatory proteins, MinC and Cdv3, enables control over cell morphology by disrupting FtsZ localization and cell division without preventing continued cell growth. By varying the expression of these proteins, we can tune the lengths of cyanobacterial cells across a broad dynamic range, anywhere from an ∼20% increased length (relative to the wild type) to near-millimeter lengths. Highly elongated cells exhibit increased rates of sedimentation under low centrifugal forces or by gravity-assisted settling. Furthermore, hyperelongated cells are also more susceptible to lysis through the application of mild physical stress. Collectively, these results demonstrate a novel approach toward decreasing harvesting and processing costs associated with mass cyanobacterial cultivation by altering morphology at the cellular level.IMPORTANCE We show that the cell length of a model cyanobacterial species can be programmed by rationally manipulating the expression of protein factors that suppress cell division. In some instances, we can increase the size of these cells to near-millimeter lengths with this approach. The resulting elongated cells have favorable properties with regard to cell harvesting and lysis. Furthermore, cells treated in this manner continue to grow rapidly at time scales similar to those of uninduced controls. To our knowledge, this is the first reported example of engineering the cell morphology of cyanobacteria or algae to make them more compatible with downstream processing steps that present economic barriers to their use as alternative crop species. Therefore, our results are a promising proof-of-principle for the use of morphology engineering to increase the cost-effectiveness of the mass cultivation of cyanobacteria for various sustainability initiatives.


Assuntos
Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Engenharia Genética , Synechococcus/citologia , Synechococcus/crescimento & desenvolvimento , Bacteriólise , Biomassa , Centrifugação , Synechococcus/genética
9.
J Am Chem Soc ; 138(16): 5262-70, 2016 04 27.
Artigo em Inglês | MEDLINE | ID: mdl-26704697

RESUMO

Bacterial microcompartments (BMCs) are self-assembling organelles composed of a selectively permeable protein shell and encapsulated enzymes. They are considered promising templates for the engineering of designed bionanoreactors for biotechnology. In particular, encapsulation of oxidoreductive reactions requiring electron transfer between the lumen of the BMC and the cytosol relies on the ability to conduct electrons across the shell. We determined the crystal structure of a component protein of a synthetic BMC shell, which informed the rational design of a [4Fe-4S] cluster-binding site in its pore. We also solved the structure of the [4Fe-4S] cluster-bound, engineered protein to 1.8 Å resolution, providing the first structure of a BMC shell protein containing a metal center. The [4Fe-4S] cluster was characterized by optical and EPR spectroscopies; it has a reduction potential of -370 mV vs the standard hydrogen electrode (SHE) and is stable through redox cycling. This remarkable stability may be attributable to the hydrogen-bonding network provided by the main chain of the protein scaffold. The properties of the [4Fe-4S] cluster resemble those in low-potential bacterial ferredoxins, while its ligation to three cysteine residues is reminiscent of enzymes such as aconitase and radical S-adenosymethionine (SAM) enzymes. This engineered shell protein provides the foundation for conferring electron-transfer functionality to BMC shells.


Assuntos
Proteínas Ferro-Enxofre/metabolismo , Engenharia de Proteínas/métodos , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Cristalografia por Raios X , Cisteína/química , Espectroscopia de Ressonância de Spin Eletrônica , Proteínas Ferro-Enxofre/química , Oxirredução
10.
Sci Rep ; 14(1): 13823, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38879676

RESUMO

Exoplanet atmospheres are expected to vary significantly in thickness and chemical composition, leading to a continuum of differences in surface pressure and atmospheric density. This variability is exemplified within our Solar System, where the four rocky planets exhibit surface pressures ranging from 1 nPa on Mercury to 9.2 MPa on Venus. The direct effects and potential challenges of atmospheric pressure and density on life have rarely been discussed. For instance, atmospheric density directly affects the possibility of active flight in organisms, a critical factor since without it, dispersing across extensive and inhospitable terrains becomes a major limitation for the expansion of complex life. In this paper, we propose the existence of a critical atmospheric density threshold below which active flight is unfeasible, significantly impacting biosphere development. To qualitatively assess this threshold and differentiate it from energy availability constraints, we analyze the limits of active flight on Earth, using the common fruit fly, Drosophila melanogaster, as a model organism. We subjected Drosophila melanogaster to various atmospheric density scenarios and reviewed previous data on flight limitations. Our observations show that flies in an N2-enriched environment recover active flying abilities more efficiently than those in a helium-enriched environment, highlighting behavioral differences attributable to atmospheric density vs. oxygen deprivation.

11.
Sci Total Environ ; 912: 168832, 2024 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-38036131

RESUMO

The aptamer (Apt) and the molecularly imprinted polymer (MIP), as effective substitutes for antibodies, have received widespread attention from researchers because of their creation. However, the low stability of Apt in harsh detection environment and the poor specificity of MIP have hindered their development. Therefore, some researchers have attempted to combine MIP with Apt to explore whether the effect of "1 + 1 > 2" can be achieved. Since its first report in 2013, MIP-Apt dual recognition elements have become a highly focused research direction in the fields of biology and chemistry. MIP-Apt dual recognition elements not only possess the high specificity of Apt and the high stability of MIP in harsh detection environment, but also have high sensitivity and affinity. They have been successfully applied in medical diagnosis, food safety, and environmental monitoring fields. This article provides a systematic overview of three preparation methods for MIP-Apt dual recognition elements and their application in eight different types of sensors. It also provides effective insights into the problems and development directions faced by MIP-Apt dual recognition elements.


Assuntos
Impressão Molecular , Polímeros Molecularmente Impressos , Inocuidade dos Alimentos , Impressão Molecular/métodos
12.
Anal Chim Acta ; 1318: 342922, 2024 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-39067914

RESUMO

BACKGROUND: Due to its wide application, procymidone has become one of the pesticides with high detection rates in supervision and sampling. Therefore, it is necessary to establish a rapid and efficient method for the detection of procymidone. However, an important bottleneck restricting the development of rapid detection methods of procymidone is that its specific recognition elements are rarely reported. In this work, Capture-SELEX and post-SELEX were used in aptamer screening, and the obtained aptamers were used to construct an aptamer-based lateral flow assay (LFA). RESULTS: Firstly, a specific aptamer Seq15 was obtained for procymidone by Capture-SELEX, and its dissociation constant (Kd) was 24.22 nM. Secondly, post-SELEX was used to analyze and modify Seq15 to improve its performance, and the Kd of the truncated sequence Seq15-2 was 21.28 nM. In addition to this, the broad-specificity aptamer Seq17-1 was obtained via post-SELEX. Seq17-1 could broadly recognize dicarboximide fungicides (procymidone, iprodione, chlozolinate, dimethachlon and vinclozolin) and their metabolic derivative (3,5-dichloroaniline). Finally, the specific aptamer-based LFA of procymidone was constructed, and the limit of detection (LOD) was 0.79 ng/mL. Meanwhile, the LODs of dicarboximide fungicides and their metabolic derivative were 0.62, 0.64, 0.71, 0.69, 0.64 and 0.66 ng/mL, respectively. The above LFAs were highly specific and stable, and had been successfully used for the detection of vegetable samples. SIGNIFICANCE: Under the combination of Capture-SELEX and Post-SELEX, this study not only provides specific recognition elements for rapid detection of procymidone, but also provides new ideas for the discovery of broad-specificity aptamers. Combining broad-specificity primary detection and single-specificity quantification, a composite aptamer-based LFA detection platform has been developed, which significantly improves detection efficiency.


Assuntos
Aptâmeros de Nucleotídeos , Técnica de Seleção de Aptâmeros , Aptâmeros de Nucleotídeos/química , Técnica de Seleção de Aptâmeros/métodos , Fungicidas Industriais/análise , Limite de Detecção
13.
Talanta ; 277: 126443, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38897007

RESUMO

Stable detection of diazinon (DZN) residues in vegetables is important for food safety. In this work, an electrochemiluminescence (ECL) aptasensor with dual-catalytic glucose in-situ production of H2O2 was constructed for the stable detection of DZN in vegetables. Firstly, MWCNTs@MB was prepared using π-π stacking interactions between methylene blue (MB) and multi-walled carbon nanotubes (MWCNTs) to enhance the loading of MB on an electrode and thus catalyze the generation of H2O2 from glucose. Secondly, Cu2O@AuNPs was formed by loading AuNPs on the surface of Cu2O through spontaneous reduction reaction, which improved the interfacial charge transfer, Cu2O nano-enzyme had glucose oxidase mimicking activity and could further catalyze the production of more H2O2 from glucose. MWCNTs@MB and Cu2O@AuNPs played a key role in the in-situ generation of co-reacting reagent H2O2, which solved the problem of unstable detection caused by the easy decomposition of the H2O2 solution added to the luminescence system. In addition, the aptamer was immobilized on the electrode surface by forming Au-S bonds with Cu2O@AuNPs. As a result, the ECL aptasensor performed good linearity in 1.00 pg mL-1-1.00 µg mL-1 and a low limit of detection (LOD) to 0.39 pg mL-1 (S/N = 3). This work provided an effective method for the accurate and stable detection of DZN residues in vegetables, which was of great significance in ensuring food safety and assessing the environmental risk of DZN.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Diazinon , Técnicas Eletroquímicas , Glucose , Ouro , Peróxido de Hidrogênio , Medições Luminescentes , Nanotubos de Carbono , Verduras , Peróxido de Hidrogênio/química , Verduras/química , Glucose/análise , Glucose/química , Técnicas Eletroquímicas/métodos , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Medições Luminescentes/métodos , Ouro/química , Nanotubos de Carbono/química , Diazinon/análise , Diazinon/química , Nanopartículas Metálicas/química , Cobre/química , Catálise , Eletrodos , Glucose Oxidase/química , Glucose Oxidase/metabolismo , Resíduos de Praguicidas/análise , Resíduos de Praguicidas/química , Limite de Detecção , Contaminação de Alimentos/análise , Azul de Metileno/química
14.
Int J Food Microbiol ; 423: 110831, 2024 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-39083880

RESUMO

In this study, a multi-scale attention transformer (MSAT) was coupled with hyperspectral imaging for classifying peanut kernels contaminated with diverse Aspergillus flavus fungi. The results underscored that the MSAT significantly outperformed classic deep learning models, due to its sophisticated multi-scale attention mechanism which enhanced its classification capabilities. The multi-scale attention mechanism was utilized by employing several multi-head attention layers to focus on both fine-scale and broad-scale features. It also integrated a series of scale processing layers to capture features at different resolutions and incorporated a self-attention mechanism to integrate information across different levels. The MSAT model achieved outstanding performance in different classification tasks, particularly in distinguishing healthy peanut kernels from those contaminated with aflatoxigenic fungi, with test accuracy achieving 98.42±0.22%. However, it faced challenges in differentiating peanut kernels contaminated with aflatoxigenic fungi from those with non-aflatoxigenic contamination. Visualization of attention weights explicitly revealed that the MSAT model's multi-scale attention mechanism progressively refined its focus from broad spatial-spectral features to more specialized signatures. Overall, the MSAT model's advanced processing capabilities marked a notable advancement in the field of food quality safety, offering a robust and reliable tool for the rapid and accurate detection of Aspergillus flavus contaminations in food.


Assuntos
Arachis , Aspergillus flavus , Contaminação de Alimentos , Microbiologia de Alimentos , Aspergillus flavus/isolamento & purificação , Arachis/microbiologia , Contaminação de Alimentos/análise , Inocuidade dos Alimentos , Aflatoxinas/análise , Imageamento Hiperespectral/métodos
15.
J Hazard Mater ; 477: 135296, 2024 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-39059293

RESUMO

A lateral flow immunoassay strip (LFIAS) is one of the most frequently rapid test technologies for carbofuran (CAR). Nevertheless, the LFIAS has a poor quantitative capability and low sensitivity. And, it also requires often complex sample handling steps, making testing time longer. In this study, Fe3O4 nanoparticles were successively modified with MIL-100(Fe)-based metal-organic framework (MOF) and chloroplatinic acid hexahydrate to obtain a core-shell complex of Fe3O4-MOF-Pt. The complex had a peroxidase-mimicking activity catalytic function that enabled signal amplification and sensitivity enhancement. Upon coupling with carbofuran monoclonal antibody (CAR-mAb), the magnetic separation properties of the probe enabled target-specific enrichment. The LFIAS based on Fe3O4-MOF-Pt nanocomposites could detect CAR in the range of 0.25-50 ng mL-1 with a limit of detection (LOD) of 0.15 ng mL-1, enabling colorimetric and catalytic analysis. In addition, the method showed high specificity and stability for detecting CAR in various vegetables, and recovery rates of the spiked samples were 91.40%-102.40%. In conclusion, this study provided one-stop detection of "target enrichment-visual inspection". While lowering the LOD, it reduced the detection time and improved the detection efficiency. The multifunctional Fe3O4-MOF-Pt nanocomposite provides an idea for the construction of novel multifunctional probes to improve the detection performance of conventional LFIAS.


Assuntos
Carbofurano , Limite de Detecção , Verduras , Carbofurano/análise , Verduras/química , Imunoensaio/métodos , Contaminação de Alimentos/análise , Estruturas Metalorgânicas/química , Platina/química , Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Inseticidas/análise , Nanocompostos/química , Nanopartículas de Magnetita/química
16.
J Hazard Mater ; 476: 134935, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-38905980

RESUMO

Time-resolved fluorescent lateral immunoassay strip (TRFLIS) is a reliable and rapid method for detecting acetamiprid. However, its sensitivity is often affected by the structural patterns and stability of the fluorescent probe. Researchers have shown significant interests in using goat anti-mouse IgG (GaMIgG) which is indirectly bound to time-resolved fluorescent microsphere (TRFM) and antibody. This allowed for oriented modification of the antibody. However, the stability of fluorescent probe in this binding mode remained unexplored. Herein, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride was innovatively used as a cross-linking agent to enhance the binding of antibody to GaMIgG, which improved the stability of the fluorescent probe. Under optimal working conditions, this strategy exhibited a wide linear response range of 5-700 ng/mL. Its limit of detection (LOD) was 0.62 ng/mL, the visual LOD was 5 ng/mL, and the limit of quantification (LOQ) of 2.06 ng/mL. Additionally, under tomato matrix, leek matrix and Chinese cabbage matrix, the linear response ranges were 5-400, 5-300, and 5-700 ng/mL, with LODs of 0.16, 0.60, and 0.41 ng/mL, with LOQs of 0.53, 2.01 and 1.37 ng/mL, respectively. In conclusion, this strategy effectively reduced the dosage of acetamiprid antibody compared with TRFM directly linking acetamiprid antibody, and greatly increased the sensitivity of TRFLIS. Meanwhile, it demonstrated outstanding specificity and accuracy in acetamiprid detection and had been successfully applied to vegetable samples. This method enables rapid and accurate detection of large-volume samples by combining qualitative and quantitative methods. As such, it has great potential in the development of low-cost and high-performance immunochromatographic platforms.


Assuntos
Corantes Fluorescentes , Limite de Detecção , Neonicotinoides , Neonicotinoides/análise , Corantes Fluorescentes/química , Imunoensaio/métodos , Animais , Reagentes de Ligações Cruzadas/química , Contaminação de Alimentos/análise , Inseticidas/análise , Anticorpos/química , Anticorpos/imunologia , Brassica/química , Camundongos , Solanum lycopersicum/química
17.
Food Chem ; 458: 140330, 2024 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-38970953

RESUMO

Food safety is an important cornerstone of protecting human health and life. Therefore, it is of great significance to detect possible pollutants in food sensitively and efficiently. Molecularly imprinted polymers (MIPs) and metal-organic frameworks (MOFs) have been widely used in the adsorption and detection of food pollutants. However, traditional MIPs have problems such as uneven loading of the imprinted cavity and slow mass transfer efficiency. While the adsorption of MOFs has low specificity and cannot accurately identify target molecules. Therefore, some researchers have taken advantage of the high specific recognition abilities of MIPs and the large specific surface areas, high porosity and easy functionalization of MOFs to combine MOFs with MIPs, and have achieved a series of important results in the field of food safety detection. This paper reviews the research progress of the application of MOFs-MIPs in the field of food safety detection from 2019 to 2024. It furnishes researchers interested in this domain with a rapid and comprehensive grasp of the latest research status, it also offers them a chance to anticipate future development trends, thereby supporting the continuous advances of MOFs-MIPs in food safety detection.


Assuntos
Contaminação de Alimentos , Inocuidade dos Alimentos , Estruturas Metalorgânicas , Polímeros Molecularmente Impressos , Estruturas Metalorgânicas/química , Polímeros Molecularmente Impressos/química , Contaminação de Alimentos/análise , Adsorção , Impressão Molecular , Humanos , Polímeros/química
18.
J Hazard Mater ; 476: 135112, 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-38981234

RESUMO

In light of the significant risks that mycotoxins posed to public health and environmental safety, this research developed an adsorbent MIPs/Apt/AuNPs@ZIF-67 (MA-AZ) utilizing a dual-recognition approach combining molecularly imprinted polymers (MIPs) and aptamer (Apt). This innovative method enabled the effective and highly selective recognition and enrichment of ochratoxin A (OTA). ZIF-67 was utilized as a carrier with a substantial specific surface area, and gold nanoparticles (AuNPs) were loaded on its surface to fix the thiol-modified Apt on the surface of the carrier. Then, an initiator was used to initiate a polymerization reaction, and the generated MIPs coated Apt/AuNPs@ZIF-67, thereby synthesizing the MA-AZ with a "synergistic recognition" effect. The Apt significantly increased the number of recognition sites within the imprinted cavities, and MIPs played roles in identifying targets, fixing and protecting Apt. The combination of the both produced the effect of "1+1>2". The study on the adsorption performance of MA-AZ found that the adsorption capacity of MA-AZ could reach 65.1 mg/g, and the imprinted factor was 5.48. In addition, MA-AZ exhibited excellent stability, specificity, reusability and recovery rate. Thus, this study offers valuable insights for the recognition and enrichment of hazardous substances, and helps to promote the rapid development of safety detection.


Assuntos
Aptâmeros de Nucleotídeos , Ouro , Nanopartículas Metálicas , Polímeros Molecularmente Impressos , Ocratoxinas , Ocratoxinas/química , Ocratoxinas/análise , Aptâmeros de Nucleotídeos/química , Adsorção , Polímeros Molecularmente Impressos/química , Nanopartículas Metálicas/química , Ouro/química , Impressão Molecular , Limite de Detecção , Extração em Fase Sólida/métodos
19.
Anal Chim Acta ; 1287: 341938, 2024 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-38182333

RESUMO

Pathogenic bacteria are primarily kinds of food hazards that provoke serious harm to human health via contaminated or spoiled food. Given that pathogenic bacteria continue to reproduce and expand once they contaminate food, pathogenic bacteria of high concentration triggers more serious losses and detriments. Hence, it is essential to detect low-dose pollution at an early stage with high sensitivity. Aptamers, also known as "chemical antibodies", are oligonucleotide sequences that have attracted much attention owing to their merits of non-toxicity, small size, variable structure as well as easy modification of functional group. Aptamer-based bioanalysis has occupied a critical position in the field of rapid detection of pathogenic bacteria. This is attributed to the unique advantage of using aptamers as recognition elements in signal amplification strategies. The signal amplification strategy is an effective means to improve the detection sensitivity. Some diverse signal amplification strategies emphasize the synthesis and assembly of nanomaterials with signal amplification capabilities, while others introduce various nucleic acid amplification techniques into the detection system. This review focuses on a variety of signal amplification strategies employed in aptamer-based detection approaches to pathogenic bacteria. Meanwhile, we provided a detailed introduction to the design principles and characteristics of signal amplification strategies, as well as the improvement of sensor sensitivity. Ultimately, the existing issues and development trends of applying signal amplification strategies in apta-sensing analysis of pathogenic bacteria are critically proposed and prospected. Overall, this review discusses from a new perspective and is expected to contribute to the further development of this field.


Assuntos
Anticorpos , Nanoestruturas , Humanos , Bactérias/genética , Poluição Ambiental , Técnicas de Amplificação de Ácido Nucleico , Oligonucleotídeos
20.
Talanta ; 269: 125471, 2024 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-38061203

RESUMO

Gold nanoparticles (AuNPs)@N-(4-aminobutyl)-N-ethylisoluminol (ABEI)@Titanium dioxide nanorods (TiO2NRs) were used as sensing materials to produce a unique encapsulated nanostructure aptasensor for the detection of acetamiprid residues in this work. ABEI, an analog of luminol, was extensively used as an electrochemiluminescence (ECL) reagent. The ECL mechanism of ABEI- hydrogen peroxide (H2O2) system had connections to a number of oxygen-centered free radicals. TiO2NRs improved ECL response with high electron transfer and a specific surface area. AuNPs were easy to biolabel and could catalyze H2O2 to enhance ECL signal. AuNPs were wrapped around TiO2NRs by utilizing the reduction property of ABEI to form wrapped modified nanomaterials. The sulfhydryl-modified aptamer bound to the nanomaterial by forming gold-sulfur (Au-S) bonds. The aptamer selectively bound to its target with the addition of acetamiprid, which caused a considerable decrease in ECL intensity and enabled quantitative detection of acetamiprid. The aptasensor showed good stability, repeatability and specificity with a broad detection range (1×10-2-1×103 nM) and a lower limit of detection (3 pM) for acetamiprid residues in vegetables. Overall, this aptasensor presents a simple and highly sensitive method for ECL detecting acetamiprid, with potential applications in vegetable safety monitoring.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Nanotubos , Ouro/química , Verduras , Nanopartículas Metálicas/química , Limite de Detecção , Peróxido de Hidrogênio/química , Medições Luminescentes/métodos , Técnicas Biossensoriais/métodos , Luminol/química , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos
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