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1.
Nanotechnology ; 2024 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-39025079

RESUMO

Novel graphene-like nanomaterials with a non-zero bandgap are important for the design of gas sensors. The selectivity toward specific targets can be tuned by introducing appropriate functional groups on their surfaces. In this study, we use first-principles simulations to investigate the covalent functionalization of single-layer graphitized BC6N with azides to yield aziridine-functionalized adducts and explore their possible use to realize ammonia sensors. First, we determine the most favorable sites for physical adsorption and chemical reaction of methylnitrene, arising from the decomposition of methylazide, onto a BC6N monolayer. Then, we examine the thermodynamics of the [1+2]-cycloaddition reaction of various phenylnitrenes and perfluorinated phenylnitrenes para-substituted with (R = CO2H, SO3H) groups, demonstrating a favorable energetics. We also monitor the effect of the functionalization on the electronic properties of the nanosheets via density of states (DOS) and band structure analyses. Finally, we test four dBC6N to gBC6N substrates in the sensing of ammonia. We show that, thanks to their hydrogen bonding capabilities, the functionalized BC6N can selectively detect ammonia, with interaction energies varying from -0.541 eV to -1.371 eV, even in presence of competing gas such as CO2 and H2O, as also confirmed by analyzing the change in the electronic properties and the values of recovery times near ambient temperature. Importantly, we model the conductance of a selected substrate alone and in presence of NH3 to determine its effect on the integrated current, showing that humidity and coverage conditions should be properly tuned to use HO3S-functionalized BC6N-based nanomaterials to develop selective gas sensors for ammonia. .

2.
Inorg Chem ; 63(18): 8059-8069, 2024 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-38655576

RESUMO

Ionic liquids (ILs) have significant potential for eco-friendly extraction of uranium from aqueous solutions, which is critical for nuclear technology, fuel cycle management, and environmental protection. This study examines the impact of the adjustable hydrophobic/hydrophilic properties of ILs on the removal of uranium(VI) (UO22+) from aqueous solutions utilizing both a novel hydrophilic IL (1-butoxyethyl-1-methylmorpholinium butoxyethylphosphite - Mor1-2O4-BOEP) and 1-heptyl-1-methylmorpholinium heptylphosphite (Mor1-7-HP) as an example of a hydrophobic IL with a similar structure. The transfer mechanism of uranyl ions from water to organic or solid phases closely depends on the physicochemical properties of ILs, especially their hydrophobicity. The hydrophobic Mor1-7-HP extracts uranyl via neutral complex formation as UO2(NO3)2-(Mor1-7-HP)2. Conversely, hydrophilic Mor1-2O4-BOEP induced selective precipitation as UO2(NO3)-(BOEP), transferring uranyl to the solid phase. Optimization of the working parameters, in terms of acidity of the aqueous solution and amount of ILs used, allowed the extraction of over 98% of U(VI). The stoichiometry of the organic complex and the precipitate was determined using physicochemical techniques. These tunable H-phosphonate-based ILs have advantages over traditional solvent extraction and conventional ILs, allowing easier handling, improved selectivity, and lower environmental impact. This work advances uranium separation techniques with applications in hydrometallurgy, particularly in the treatment of wastewater and radioactive waste for sustainable uranium recovery.

3.
Anal Chim Acta ; 1305: 342583, 2024 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-38677845

RESUMO

P-element-induced wimpy testis (PIWI)-interacting RNAs (piRNAs/piRs) are a class of small noncoding RNAs that play a crucial role in regulating various biological processes, including carcinogenesis. One specific piRNA, piR-651, has been reported to be overexpressed in both human blood serum and solid cancer tissues, that can be used a viable biomarker in cancer diagnosis. Early diagnosis of cancer can help reduce the burden of the disease and improve survival rates. In the present work, we report for the first time a smartphone-based colorimetric biosensor for highly sensitive and specific detection of piR-651 thanks to an enzymatic signal amplification, which yielded high colorimetric intensities. Indeed, a heteroduplex DNA:RNA was formed in the presence of piR-651 with the capture DNA probe immobilized on the magnetic beads for easy magnetic separation. Then, a HRP tethered to anti-DNA:RNA (S9.6) was used to reveal the DNA-RNA heteroduplex formed by catalyzing the oxidation of TMB substrate into colorimetric TMBox, which absorbs at 630 nm. The absorbance is positively proportional to the piR-651 concentrations. On the other hand, the colorimetric product of the assay can be photographed with a smartphone camera and analyzed using ImageJ software. Using a smartphone and under optimal conditions, the biosensor responded linearly to the logarithm of piRNA-651 from 8 fM to 100 pM with a detection limit of 2.3 fM and discriminates against other piRNAs. It was also successfully applied to the determination of piRNA-651 levels in spiked human serum.


Assuntos
Técnicas Biossensoriais , RNA Interferente Pequeno , Smartphone , Humanos , RNA Interferente Pequeno/química , Técnicas Biossensoriais/métodos , Colorimetria , DNA/química , Limite de Detecção , RNA de Interação com Piwi
4.
RSC Adv ; 14(17): 12071-12080, 2024 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-38628480

RESUMO

The SARS-CoV-2 (COVID-19) pandemic had a strong impact on societies and economies worldwide and tests for high-performance detection of SARS-CoV-2 biomarkers are still needed for potential future outbreaks of the disease. In this paper, we present the different steps for the design of an aptamer-based surface-enhanced Raman scattering (BioSERS) sensing chip capable of detecting the coronavirus nucleocapsid protein (N protein) in spiked phosphate-buffered solutions and real samples of human blood serum. Optimization of the preparation steps in terms of the aptamer concentration used for the functionalization of the silver nanoparticles, time for affixing the aptamer, incubation time with target protein, and insulation of the silver active surface with cysteamine, led to a sensitive BioSERS chip, which was able to detect the N protein in the range from 1 to 75 ng mL-1 in spiked phosphate-buffered solutions with a detection limit of 1 ng mL-1 within 30 min. Furthermore, the BioSERS chip was used to detect the target protein in scarcely spiked human serum. This study demonstrates the possibility of a clinical application that can improve the detection limit and accuracy of the currently commercialized SARS-CoV-2 immunodiagnostic kit. Additionally, the system is modular and can be applied to detect other proteins by only changing the aptamer.

5.
RSC Adv ; 14(12): 8007-8015, 2024 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-38454949

RESUMO

Ensuring food security is crucial for public health, and the presence of mycotoxins, produced by fungi in improperly stored processed or unprocessed food, poses a significant threat. This research introduces a novel approach - a disposable aptasensing platform designed for the detection of ochratoxin A (OTA). The platform employs gold-nanostructured screen-printed carbon electrodes functionalized with a ferrocene derivative, serving as an integrated faradaic transducing system, and an anti-OTA aptamer as a bioreceptor site. Detection relies on the ferrocene electrochemical signal changes induced by the aptamer folding in the presence of the target molecule. Remarkably sensitive, the platform detects OTA within the range of 0.5 to 70 ng mL-1 and a detection limit of 11 pg mL-1. This limit is approximately 200 times below the levels stipulated by the European Commission for agricultural commodities. Notably, the sensing device exhibits efficacy in detecting OTA in complex media, such as roasted coffee beans and wine, without the need for sample pretreatment, yielding accurate recoveries. Furthermore, while label-free electrochemical aptasensors have proliferated, this study addresses a gap in understanding the binding mechanisms of some aptasensors. To enhance the experimental findings, a theoretical study was conducted to underscore the specificity of the anti-OTA aptamer as a donor for OTA detection. The molecular docking technique was employed to unveil the key binding region of the aptamer, providing valuable insights into the aptasensor specificity.

6.
Mikrochim Acta ; 190(11): 434, 2023 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-37821740

RESUMO

An ultrasensitive capacitance-based biosensor has been developed capable of detecting the kanamycin (KAN) antibiotic at sub-femtomolar levels. The biosensor was constructed using a potential-pulse-assisted method, allowing for the layer-by-layer deposition of a melanin-like polymeric film (MLPF) on an electrode surface modified with gold nanoparticles (AuNPs). The MLPF was formed through the electrochemical polymerization of dopamine and the specific kanamycin aptamer. By optimizing the operating parameters, we achieved a label-free detection of kanamycin by monitoring the variation of pseudocapacitive properties of the MLPF-modified electrode using electrochemical impedance spectroscopy. The developed biosensor demonstrated a wide linear response ranging from 1 fM to 100 pM, with a remarkable limit of detection of 0.3 fM (S/N = 3) for kanamycin. Furthermore, the biosensor was successfully applied to detect kanamycin in milk samples, exhibiting good recovery. These findings highlight the promising potential of the aptasensor for determination of antibiotic residues and ensuring food safety. In conclusion, our ultrasensitive capacitance-based biosensor provides a reliable and efficient method for detecting trace amounts of kanamycin in dairy products. This technology can contribute to safeguarding consumer health and maintaining high food safety standards.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Canamicina , Ouro/química , Aptâmeros de Nucleotídeos/química , Nanopartículas Metálicas/química , Oxirredução , Antibacterianos , Eletrodos , Técnicas Biossensoriais/métodos
7.
R Soc Open Sci ; 10(8): 230294, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37538749

RESUMO

The development of cost-effective materials for fabricating electrodes is crucial for drug, pharmaceutical and environmental applications. This paper presents the synthesis and characterization of a novel polyketimine (PKI) membrane obtained by condensing partially of different weight percentages of oxidized polyvinyl alcohol and aminated polyether sulfone. Using the PKI membrane as a scaffold, we introduced laser-induced graphene electrodes (LIGEs) for the efficient electrochemical sensing of paracetamol (PCM), which serves as a model drug. Electrochemical measurements were conducted to assess the physico-chemical properties, including laser-induced porous graphene features, such as the heterogeneous electron transfer (HET) rate and electrochemically active surface area (ECSA). The obtained results demonstrate that the LIGEs exhibit excellent performance in PCM sensing, showing a linear detection range of 50-600 µM with a detection limit (LOD) as low as 14.3 µM and a good selectivity toward uric acid. Furthermore, the functionalization of the electrode surface with AuNPs improved the electrode physico-chemical properties (HET and ECSA) and lowered the detection limit down to 1.1 µM. Consequently, these affordable electrodes hold great potential for analysing other drugs and detecting heavy metal cations in various applications.

9.
Mikrochim Acta ; 190(8): 316, 2023 07 22.
Artigo em Inglês | MEDLINE | ID: mdl-37480385

RESUMO

The level control of biological active molecules in human body fluids is important for the surveillance of several human diseases. Dopamine (DA) and uric acid (UA) are two important biomarkers of neurological and bone diseases, respectively. Design of sensitive and cost-effective sensors for their detection is an effervescent research field. We report on the straightforward design of laser-induced graphene electrodes (LIGEs) from the laser ablation of a polyimide substrate and their modification by electrochemical deposition of gold nanoparticles (AuNPs/LIGE) and their uses as chemosensors. Electrochemical investigations showed that the presence of gold nanoclusters onto the electrode surface improved the electrochemical surface area (ECSA) and the heterogenous electron transfer (HET) rate. Furthermore, the AuNPs/LIGEs can be used to detect simultaneously low concentrations of DA and UA in presence of ascorbic acid (AA) as an potentially interfering substance at redox potentials of 300 mV, 230 mV and 450 mV and 91 mV, respectively, compared with the Ag/AgCl (3 M KCl) reference electrode in cyclic voltametric. The method displayed linear ranges varying from 2 to 20 µM and 5 to 50 µM, led to limits of detection of 0.37 µM and 0.71 µM for DA and UA, respectively. The AuNPs/LIGE was applied to simultaneously detect both analytes in scarcely diluted human serum with good recoveries. The data show that the recovery percentages ranged from 94% ± 2.1 to 102 % ± 0.5 and from 94% ±0.3 to 112% ± 1.4 for dopamine and uric acid, respectively. Thus, the AuNPs/LIGEs are promising candidates for the detection of other biologically active molecules such as drugs, pesticides, and metabolites.


Assuntos
Grafite , Nanopartículas Metálicas , Humanos , Dopamina , Ácido Úrico , Ouro , Lasers , Eletrodos
10.
RSC Adv ; 13(31): 21336-21344, 2023 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-37465569

RESUMO

Regulatory bodies play a crucial role in establishing limits for food additives to ensure food quality and safety of food products, as excessive usage poses risks to consumers. In the context of processed animal-based foodstuffs, nitrite is commonly utilized as a means to slow down bacterial degradation. In this study, we have successfully leveraged the redox activity of an electrochemically deposited polydopamine (pDA) film onto gold nanoparticle (AuNP)-modified screen-printed electrodes (SPCE) to develop a sensitive and versatile methodology for the detection of nitrite using redox capacitance spectroscopy. By exploiting the interaction of the AuNPs/pDA electroactive interface with the target nitrite ions, we observed distinct changes in the redox distribution, subsequently leading to modifications in the associated redox capacitance. This alteration enables the successful detection of nitrite, exhibiting a linear response within the concentration range of 10 to 500 µM, with a limit of detection of 1.98 µM (S/N = 3). Furthermore, we applied the developed sensor to analyze nitrite levels in processed meats, yielding good recoveries. These results demonstrate the potential of our approach as a promising method for routine detection of ions.

11.
RSC Adv ; 13(28): 19607-19616, 2023 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-37388147

RESUMO

Graphene and other 2D materials have gained significant attention in the development of gas sensors. In this study, we employed Density Functional Theory (DFT) to investigate the adsorption properties of diazomethanes (1a-1g) with various functional groups (R = OH (a), OMe (b), OEt (c), OPr (d), CF3 (e), Ph (f)) on pristine graphene. Furthermore, we explored the adsorption behavior of activated carbenes (2a-2g) generated from the decomposition of diazomethanes on graphene, as well as the functionalized graphene derivatives (3a-3g) resulting from [2 + 1] cycloaddition reactions between (2a-2g) and graphene. The interaction between these functionalized derivatives (3a-3g) and toxic gases was also investigated. Our results revealed that carbenes exhibited a stronger affinity for graphene compared to diazomethanes. The adsorption energy of esters (3b, 3c, and 3d) on graphene decreased relative to compound 3a, while 3e exhibited increased adsorption energy due to the electron-withdrawing effect of fluorine atoms. Additionally, the adsorption energy of phenyl and nitrophenyl groups (3f and 3g) decreased due to their π-stacking interaction with graphene. Importantly, all functionalized derivatives (3a-3g) demonstrated favorable interactions with gases. Notably, the derivative 3a, acting as a hydrogen bonding donor, exhibited superior performance. Furthermore, modified graphene derivatives exhibited the highest adsorption energy with NO2 gas, highlighting their potential for selective NO2 sensing applications. These findings contribute to the understanding of gas-sensing mechanisms and the design of novel graphene-based sensor platforms.

12.
J Colloid Interface Sci ; 646: 1-10, 2023 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-37178610

RESUMO

A facile and expandable methodology was successfully developed to fabricate laser-induced graphene from novel pristine aminated polyethersulfone (amPES) membranes. The as-prepared materials were applied as flexible electrodes for microsupercapacitors. The doping of amPES membranes with various weight percentages of carbon black (CB) microparticles was then performed to improve their energy storage performance. The lasing process allowed the formation of sulfur- and nitrogen-codoped graphene electrodes. The effect of electrolyte on the electrochemical performance of as-prepared electrodes was investigated and the specific capacitance was significantly enhanced in 0.5 M HClO4. Remarkably, the highest areal capacitance of 47.3 mF·cm-2 was achieved at a current density of 0.25 mA·cm-2. This capacitance is approximately 12.3 times higher than the average value for commonly used polyimide membranes. Furthermore, the energy and power densities were as high as 9.46 µWh·cm-2 and 0.3 mW·cm-2 at 0.25 mA·cm-2, respectively. The galvanostatic charge-discharge experiments confirmed the excellent performance and stability of amPES membranes during 5,000 cycles, where more than 100% of capacitance retention was achieved and the coulombic efficiency was improved up to 96.67%. Consequently, the fabricated CB-doped PES membranes offer several advantages including low carbon fingerprint, cost-effectiveness, high electrochemical performance and potential applications in wearable electronic systems.

13.
J Comput Chem ; 44(15): 1426-1436, 2023 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-36905299

RESUMO

Recently, halogen bonding (XB) has received increased attention as a new type of non-covalent interaction widely present in nature. In this work, quantum chemical calculations at DFT level have been carried out to investigate halogen bonding interactions between COn (n = 1 or 2) and dihalogen molecules XY (X = F, Cl, Br, I and Y = Cl, Br, I). Highly accurate all-electron data, estimated by CCSD(T) calculations, were used to benchmark the different levels of computational methods with the objective of finding the best accuracy/computational cost. Molecular electrostatic potential, interaction energy values, charge transfer, UV spectra, and natural bond orbital (NBO) analysis were determined to better understand the nature of the XB interaction. Density of states (DOS) and projected DOS were also computed. Hence, according to these results, the magnitude of the halogen bonding is affected by the halogen polarizability and electronegativity, where for the more polarizable and less electronegative halogen atoms, the σ-hole is bigger. Furthermore, for the halogen-bonded complexes involving CO and XY, the OC∙∙∙XY interaction is stronger than the CO∙∙∙XY interaction. Thus, the results presented here can establish fundamental characteristics of halogen bonding in media, which would be very helpful for applying this noncovalent interaction for the sustainable capture of carbon oxides.

14.
Talanta ; 258: 124445, 2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-36924636

RESUMO

In silico evaluation of aptamer/target interactions can facilitate the development of efficient biosensor with high specificity and affinity. In this work, we present in silico, i.e. structural similarity, molecular docking and molecular dynamics selection of the aptamer with sufficient binding properties for acetamiprid (ACE), a nicotine-like pesticide, and its use to design aptamer-modified magnetic beads bearing ferrocene co-immobilized label for capacitive detection of ACE. Taking advantages of the aptamer higher stability and binding affinity, the specific properties of magnetic beads and the redox properties of ferrocene moiety, the developed aptasensor showed promising analytical performances for ACE detection, using electrochemical capacitance spectroscopy, with a linear response ranging from 1 fM to 100 pM and a limit of detection of 0.94 fM (S/N = 3). Furthermore, it was successfully applied to detect ACE in fortified tomatoes samples, proving a promising approach for routine detection of pesticide in real agricultural samples.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Praguicidas , Simulação de Acoplamento Molecular , Metalocenos/química , Aptâmeros de Nucleotídeos/química , Praguicidas/análise , Técnicas Biossensoriais/métodos , Fenômenos Magnéticos , Limite de Detecção , Técnicas Eletroquímicas/métodos
15.
Biosensors (Basel) ; 13(2)2023 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-36832006

RESUMO

Point mutations are common in the human DNA genome and are closely related to higher susceptibility to cancer diseases. Therefore, suitable methods for their sensing are of general interest. In this work, we report on a magnetic electrochemical bioassay using DNA probes tethered to streptavidin magnetic beads (strep-MBs) to detect T > G single nucleotide polymorphism (SNP) within the inteleukin-6 (IL6) gene in human genomic DNA. In the presence of the target DNA fragment and tetramethylbenzidine (TMB), the electrochemical signal related to the oxidation of TMB is observed, which is much higher than the one obtained in the absence of the target. The key parameters affecting the analytical signal, such as the concentration of the biotinylated probe, its incubation time with strep-MBs, DNA hybridization time, and TMB loading, were optimized using the electrochemical signal intensity and signal-to-blank (S/B) ratio as selection criteria. Using spiked buffer solutions, the bioassay can detect the mutated allele in a wide range of concentrations (over six decades) with a low detection limit (7.3 fM). Furthermore, the bioassay displays a high specificity with high concentrations of the major allele (one mismatched), and two mismatched and non-complementary DNA. More importantly, the bioassay can detect the variation in scarcely diluted human DNA, collected from 23 donors, and can reliably distinguish between heterozygous (TG genotype) and homozygous (GG genotype) in respect to the control subjects (TT genotype), where the differences are statistically highly significant (p-value < 0.001). Thus, the bioassay is useful for cohort studies targeting one or more mutations in human DNA.


Assuntos
Técnicas Biossensoriais , Interleucina-6 , Humanos , Mutação Puntual , DNA , Hibridização de Ácido Nucleico/métodos , Sondas de DNA , Estreptavidina , Técnicas Biossensoriais/métodos
16.
Biosensors (Basel) ; 13(2)2023 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-36832065

RESUMO

Rapid and specific detection of pathogenic bacteria in fecal samples is of critical importance for the diagnosis of neonatal diarrhea in veterinary clinics. Nanobodies are a promising tool for the treatment and diagnosis of infectious diseases due to their unique recognition properties. In this study, we report the design of a nanobody-based magnetofluorescent immunoassay for the sensitive detection of pathogenic Escherichia coli F17-positive strains (E. coli F17). For this, a camel was immunized with purified F17A protein from F17 fimbriae and a nanobody library was constructed by phage display. Two specific anti-F17A nanobodies (Nbs) were selected to design the bioassay. The first one (Nb1) was conjugated to magnetic beads (MBs) to form a complex capable of efficiently capturing the target bacteria. A second horseradish peroxidase (HRP)-conjugated nanobody (Nb4) was used for detection by oxidizing o-phenylenediamine (OPD) to fluorescent 2,3-diaminophenazine (DAP). Our results show that the immunoassay recognizes E. coli F17 with high specificity and sensitivity, with a detection limit of 1.8 CFU/mL in only 90 min. Furthermore, we showed that the immunoassay can be applied to fecal samples without pretreatment and remains stable for at least one month when stored at 4 °C.


Assuntos
Escherichia coli , Anticorpos de Domínio Único , Escherichia coli/metabolismo , Anticorpos de Domínio Único/metabolismo , Imunoensaio , Ensaio de Imunoadsorção Enzimática
17.
J Pharm Biomed Anal ; 226: 115251, 2023 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-36657353

RESUMO

Tau protein is a promising biomarker for early diagnosis of Alzheimer's disease. Therefore, there is an urgent need to develop a simple and effective method for its detection. To this end, an innovative sensing device was developed using a carbon screen-printed electrode (C-SPE) decorated with graphene oxide/Prussian Blue nanocubes (GO/PBNCs) for the selective and sensitive determination of Tau-441 protein. The molecular imprinting polymer (MIP) was built on the GO/PBNCs/C-SPE by electropolymerizing 3-aminophenol (3-AMP) in the presence of the target protein using chronoamperometry, and the template was subsequently removed from the polymer matrix with oxalic acid. In parallel, a non-imprinted material (NIP) was also prepared in the absence of the target for comparison purposes. Scanning electron microscopy and transmission electron microscopy, were used to study the morphology of the modified electrode and electrochemical techniques were used to monitor the stepwise assembly of the sensor. Under optimized conditions, the sensing platform exhibited a linear range within 1.09 and 2.18 nmol/L and a detection limit of 0.01 pmol/L in spiked phosphate buffer solution (PBS). The MIP sensor showed minimal interference with uric acid and bovine albumin. The simplicity of production, affordable cost and promising performance make this sensor a potential strategic sensing platform for the detection of chemical and biological molecules.


Assuntos
Técnicas Biossensoriais , Impressão Molecular , Animais , Bovinos , Proteínas tau , Biomimética , Carbono/química , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Impressão Molecular/métodos , Eletrodos , Polímeros/química , Limite de Detecção
18.
Mikrochim Acta ; 190(1): 15, 2022 12 07.
Artigo em Inglês | MEDLINE | ID: mdl-36479645

RESUMO

Two electrochemical bioplatforms were prepared based on thiolated hairpin DNA probes tethered to AuNP-modified screen-printed electrodes to detect T > G and T > C polymorphisms, namely rs1880269 and rs1800469, present the interleukin-6 (IL6) and transforming growth factor ß1 (TGFß1) genes. The electrochemical readout was ensured by the detection of the double-stranded DNA using methylene blue as a redox probe after treatment by EcoRI restrictase. The main parameters influencing the analytical response such as the thiolated DNA probe concentration, incubation time with electrode, DNA hybridization time, EcoRI enzyme load, and its cleavage time were optimized based on the current intensity and signal-to-blank (S/B) ratio as selection criteria. Using spiked buffer solutions, the IL6 and TGFß1 E-bioplatforms display wide ranges of linearity (1 × 102-1 × 108 fM and 5 × 101-1 × 105 fM, respectively) and limits of detection (47.9 fM and 16.6 fM, respectively). The two bioelectrodes have also good discrimination toward 1-mismatched, two mismatched, and non-complementary sequences, when they were used 30-fold higher than the target sequences. More importantly, the two bioplatforms successfully detected the single nucleotide polymorphisms (SNPs) in scarcely diluted genomic DNA, collected from 52 donors, and showed they can reliably distinguish between heterozygous (TG and TC genotypes) and homozygous (GG and CC genotypes) patients with  respect to the control subjects (TT genotype), where the differences are statistically highly significant (p-value < 0.0001). Thus, the designed devices could be used to conduct large cohort studies targeting these mutations or extended to other SNPs.


Assuntos
Interleucina-6 , Neoplasias Ovarianas , Fator de Crescimento Transformador beta1 , Feminino , Humanos , Desoxirribonuclease EcoRI , DNA/genética , Ouro , Interleucina-6/genética , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/genética , Eletrodos , Fator de Crescimento Transformador beta1/genética , Análise Mutacional de DNA , Polimorfismo de Nucleotídeo Único
19.
Anal Chim Acta ; 1231: 340397, 2022 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-36220288

RESUMO

Potassium ion channels are expressed on the cell membranes, implicated in wide variety of cell functions and intimately linked to cancer cell behaviors. This work reports the first bioplatform described to date allowing simple and rapid detection of ion channel activity and the effect of their inhibitors in cancer cells. The methodology involves interrogation of the channel of interest from cells specifically captured on magnetic immunoconjugates using specific detection antibodies that are labeled with horseradish peroxidase enzyme. The channel activity is reflected by an amperometric signal transduction of the resulting magnetic bioconjugates onto screen-printed carbon electrodes. The bioplatform feasibility was proven for the detection of the Kv channels in U87 human glioblastoma cells and their blocking by scorpion venom KAaH1 and KAaH2 peptides. The obtained results confirm the high sensitivity (detection of 5 U87 cells⋅mL-1 and 0.06 µg mL-1 of KAaH2) of the proposed bioplatform and their versatility to detect both potassium channel activity and their potential inhibitors, in a given cancer cell line, with high sensitivity in a simple and fast way. This bioplatform presents potential applications in cancer and theranostic of channelopathies.


Assuntos
Imunoconjugados , Neoplasias , Venenos de Escorpião , Carbono , Peroxidase do Rábano Silvestre , Humanos , Canais Iônicos , Neoplasias/tratamento farmacológico , Peptídeos , Bloqueadores dos Canais de Potássio/farmacologia , Bloqueadores dos Canais de Potássio/uso terapêutico , Canais de Potássio , Venenos de Escorpião/farmacologia
20.
Int J Mol Sci ; 23(11)2022 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-35682707

RESUMO

Bacterial diseases cause tremendous economic losses due to high morbidity and mortality in livestock animals. F17A protein, the major subunit of F17 fimbriae, is one of the most prevalent and crucial virulence factors among the pathogenic Escherichia coli (E. coli) isolated from diarrheic and septicemic animals of various species. Purification and detection of this protein is regarded as an interesting field of investigation due to its important role as a therapeutic target, such as vaccines, and as a diagnostic tool. In this context, polyclonal rabbit antibodies recognizing F17A protein (anti-F17A antibody) were developed and used for its detection. In fact, sandwich biosensor using anti-F17A/gold nanoparticles conjugates as capture probe and anti-F17A antibody labelled with horseradish peroxidase as signal amplification probe was developed for electrochemical and fluorescent detection of purified F17A protein and live F17-positive E. coli bacteria. Good specificity and sensitivity for detection of F17-positive E. coli strains were obtained. The dynamic range for the biosensor varies from 1 × 102 to 1 × 109 CFU·mL-1 (R2 = 0.998) and the detection limit (LOD) and the IC50 value were estimated to be 37 CFU·mL-1 and 75 CFU·mL-1, respectively.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Animais , Escherichia coli/química , Ouro/química , Imunoensaio , Nanopartículas Metálicas/química , Coelhos
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