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An ultrasensitive competitive-type electrochemiluminescence immunoassay for the mycotoxin zearalenone is described. The method is based on the use of (a) a mimotope peptide that was selected from a phage displayed peptide library and used to substitute ZEN for designing the competitive assay; (b) NiFe2O4 nanotubes with large specific surface area loaded with the ECL probe Ru(bpy)32+; and (c) poly(vinylpyrrolidone) (PVP)-assisted synthesis of TiO2 mesocrystals that acts as the sensing platform and support for antibody immobilization. Under the optimized conditions and at an ECL working potential of 1.1 V, a linear response is found for ZEN in the 0.1 to 1.0 × 10-5 ng·mL-1 concentration range with a detection limit as low as 3.3 fg·mL-1. Graphical abstract An ultrasensitive competitive-type electrochemiluminescence (ECL) immunosensor based on mimotope peptide was constructed for the detection of Zearalenone.
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The Middle East respiratory syndrome corona virus (MERS-CoV) is highly pathogenic. An immunosensor for the determination of MERS-CoV is described here. It is based on a competitive assay carried out on an array of carbon electrodes (DEP) modified with gold nanoparticles. Recombinant spike protein S1 was used as a biomarker for MERS CoV. The electrode array enables multiplexed detection of different CoVs. The biosensor is based on indirect competition between free virus in the sample and immobilized MERS-CoV protein for a fixed concentration of antibody added to the sample. Voltammetric response is detected by monitoring the change in the peak current (typically acquired at a working potential of -0.05 V vs. Ag/AgCl) after addition of different concentrations of antigen against MERS-CoV. Electrochemical measurements using ferrocyanide/ferricyanide as a probe were recorded using square wave voltammetry (SWV). Good linear response between the sensor response and the concentrations from 0.001 to 100 ng.mL-1 and 0.01 to 10,000 ng.mL-1 were observed for MERS-CoV and HCoV, respectively. The assay was performed in 20 min with detection limit as low as 0.4 and 1.0 pg.mL-1 for HCoV and MERS-CoV, respectively. The method is highly selective over non-specific proteins such as Influenza A and B. The method is single-step, sensitive and accurate. It was successfully applied to spiked nasal samples. Graphical abstract An electrochemical immunoassay is described for the Middle East Respiratory Syndrome Corona Virus (MERS-CoV). The method is based on a competitive assay carried out on a carbon array electrodes (DEP) nanostructured with gold nanoparticles. The array electrodes enable the multiplexed detection of different types of Corona Virus.
Assuntos
Infecções por Coronavirus/diagnóstico , Ouro/química , Nanopartículas Metálicas/química , Coronavírus da Síndrome Respiratória do Oriente Médio/química , Anticorpos Antivirais/metabolismo , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Eletrodos , Ferrocianetos/química , Imunoensaio/métodos , Limite de Detecção , Oxirredução , Tamanho da Partícula , Sensibilidade e Especificidade , Propriedades de SuperfícieRESUMO
A flower-like Au/Cu alloy nanocomposite (Au/Cu NFs) was synthesized and used in an electrochemiluminescence (ECL) based method for sensitive determination of the dye Sudan I. The Au-g-C3N4 nanosheets as an ECL emitter were prepared by electrostatic adsorption between gold nanoparticles and g-C3N4. They form a film on a glassy carbon electrode (GCE) and then can be connected with Sudan I antigen via gold-nitrogen bond and amidation reactions. The Au/Cu NFs combined with Sudan I antibody also via the Au-N bond and was introduced into the modified GCE by specific recognition between the antibody and the antigen. The overlap between emission spectra of the Au-g-C3N4 nanosheets and absorption spectra of Au/Cu NFs enabled the appearance of ECL resonance energy transfer process. That is, when the Sudan I analyte not present, the ECL was weakened due to absorption by the gray Au/Cu NFs on applying voltages from -1.7 V to 0 V. Conversely, the Au/Cu NFs on the GCE are reduced due to the competition for the antibody between the analyte and the antigen. A strong green ECL emission was obtained. The ECL response is linear in the 0.5 pg mL-1 to 100 ng mL-1 Sudan I concentration range, and the detection limit is 0.17 pg mL-1. Graphical abstract An Au/Cu alloy flower-like nanocomposite (Au/Cu NFs) is firstly synthesized as an acceptor to constitute an electrochemiluminescence-resonance energy transfer (ECL-RET) system for sensitive measurement of Sudan I, while Au nanoparticles (Au NPs) functionalized graphitic carbon nitride (g-C3N4) acted as a donor.
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Ligas/química , Ouro/química , Grafite/química , Imunoensaio/métodos , Nanoestruturas/química , Naftóis/análise , Nitrilas/química , Eletroquímica , Análise de Alimentos , Limite de Detecção , Luminescência , Naftóis/químicaRESUMO
Immunosensors have experienced a very significant growth in recent years, driven by the need for fast, sensitive, portable and easy-to-use devices to detect biomarkers for clinical diagnosis or to monitor organic pollutants in natural or industrial environments. Advances in the field of signal amplification using enzymatic reactions, nanomaterials such as carbon nanotubes, graphene and graphene derivatives, metallic nanoparticles (gold, silver, various oxides or metal complexes), or magnetic beads show how it is possible to improve collection, binding or transduction performances and reach the requirements for realistic clinical diagnostic or environmental control. This review presents these most recent advances; it focuses first on classical electrode substrates, then moves to carbon-based nanostructured ones including carbon nanotubes, graphene and other carbon materials, metal or metal-oxide nanoparticles, magnetic nanoparticles, dendrimers and, to finish, explore the use of ionic liquids. Analytical performances are systematically covered and compared, depending on the detection principle, but also from a chronological perspective, from 2012 to 2016 and early 2017.
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Técnicas Eletroquímicas , Técnicas Biossensoriais , Ouro , Grafite , Imunoensaio , Nanopartículas Metálicas , Nanotubos de CarbonoRESUMO
The first competitive disposable amperometric immunosensor based on gliadin-functionalized carbon/nanogold screen-printed electrodes was developed for rapid determination of celiotoxic prolamins. To date, no competitive spectrophotometric or electrochemical immunoassays have yet been successfully applied to gluten detection in processed food samples, which require the use of complex prolamin extraction solutions containing additives with denaturing, reducing and disaggregating functions. Thus, in this work, great effort was put into the optimization and performance evaluation of the immunosensor in terms of suitability as a screening tool for analysis of cereal-based food samples. For this purpose, aqueous ethanol or complex extraction mixtures, as the patented Cocktail Solution®, were proved effective in the extraction of gliadin. Good sensitivity was achieved after optimization of the immunocompetitive assay, giving limit of detection and limit of quantitation of 8 and 22 ng/ml of gliadin, respectively, for ethanol extracts. The immunosensor was proved to be suitable also for samples extracted with Cocktail Solution® after a proper dilution. Analysis of real samples of different flours proved the suitability of the immunosensing device as a powerful tool for safety assessment of raw materials used for the formulation of dietary products for celiac disease patients. This immunosensor combines good analytical performance using a very simplified set-up protocol with suitability for rapid screening analysis performed using inexpensive and portable instrumentation. Graphical abstract Depiction of the development and working principle of the competitive immunosensor.
Assuntos
Técnicas Biossensoriais/métodos , Grão Comestível/química , Análise de Alimentos/métodos , Gliadina/análise , Técnicas Biossensoriais/economia , Técnicas Biossensoriais/instrumentação , Carbono/química , Doença Celíaca/etiologia , Doença Celíaca/prevenção & controle , Eletrodos , Desenho de Equipamento , Análise de Alimentos/economia , Análise de Alimentos/instrumentação , Gliadina/química , Ouro/química , Humanos , Proteínas Imobilizadas/análise , Proteínas Imobilizadas/química , Imunoensaio/economia , Imunoensaio/instrumentação , Imunoensaio/métodos , Limite de Detecção , Nanopartículas Metálicas/químicaRESUMO
The development of new electrochemiluminescence (ECL) luminophores has become a hot research topic in the field of ECL. Metal-organic frameworks (MOFs) are widely used in ECL sensors due to their excellent ECL performance, high porosity, and abundant surface functional groups. In the work, we developed a cerium-based organic backbone as an ECL luminophor using 5,10,15,20-tetrakis(4-carboxyphenyl)-porphyrin (TCPP) as organic ligand for the detection of trenbolone (TRE). Importantly, the Ce-MOFs can effectively prevent the self-aggregation and self-quenching of TCPP, so that the ECL signal is amplified. To further improve the conductivity and antigen loading capacity of the Ce-MOFs, platinum nanosheets (Pt NPs) were modified at its surface (Pt NPs@MOFs). The Au-ZnO with good biocompatibility is used as the substrate material to load that antibody through the Au-NH2. Based on the above strategy, we constructed a competitive immunosensor to achieve a highly sensitive detection of TRE. Under suitable circumstances, it was discovered that the ECL sensor had a linear relationship with the logarithm of the TRE concentration, with a limit of detection (LOD) of 3.61 fg/mL (S/N = 3). This work provides direction for the application of organic luminescent porphyrins and their derivatives in ECL sensors.
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Herein, a sensitive immunosensor for the rapid detection of Dibutyl phthalate (DBP) small molecules was developed. Its principle is based on Au@Pt/Polyethyleneimine (PEI)-rGO and hybridization chain reaction signal amplification system. First, Au@Pt/PEI-rGO modified electrode is used as a signal amplification platform and the antibody is fixed. In the presence of target DBP, it competes with DBP-bovine serum albumin-S0 for binding antibody. Finally, the DNA concatemer carrying a large number of signal molecules methylene blue combines with its S0 to produce a reduced electrochemical signal. Under the most reasonable conditions, this immunosensor showed a good linear range from1 pg mL-1 to 0.1 µg mL-1with a low detection limit of 0.276 pg mL-1. It has been verified that this immunosensor has good selectivity, reproducibility and stability. The designed biosensing solution will open up a new path for DBP monitoring and other small molecule targets in the food field.
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Técnicas Biossensoriais , Nanopartículas Metálicas , Dibutilftalato , Técnicas Eletroquímicas , Ouro/química , Grafite , Imunoensaio , Limite de Detecção , Nanopartículas Metálicas/química , Polietilenoimina/química , Reprodutibilidade dos TestesRESUMO
A simple, rapid and robust method to quantify SARS-CoV-2 neutralizing antibodies (nAbs) is urgently needed for determining COVID-19 serodiagnosis, vaccine development and evaluation of vaccine efficacy. In this study, we report sandwich/competitive immuno-sensors based on lateral chromatography micro-interface for accurate quantification of SARS-CoV-2 nAbs. Fluorescent microspheres (FMS) labeled receptor binding domain (RBD) antigen was prepared for detection of nAbs with high sensitivity. Sandwich and competitive immunoassay were conducted on the microfluidic-based sensor within 10 min and the fluorescent signal of immunoassay was analyzed by a portable microfluidic immunoassay instrument. The nAbs detection range of sandwich immuno-sensor and competitive immuno-sensor was 4.0 ng/mL to 400 ng/mL and 2.13 ng/mL to 213 ng/mL, respectively. Furthermore, the sandwich immuno-sensor was demonstrated to be comparable with existing methods and used to detect 182 clinical serum samples from vaccinated individuals. Sandwich immuno-sensor based on lateral chromatography micro-interface allowed reliable, fast, and low-cost detection of nAbs, which holds considerable potential for nAbs testing.
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Anticorpos Neutralizantes , COVID-19 , Anticorpos Antivirais , Humanos , Imunoensaio , SARS-CoV-2RESUMO
A competitive-type PEC immunosensor for 17ß-estradiol (E2) detection was successfully fabricated using ZnIn2S4@NH2-MIL-125(Ti) composite as matrix. The excellent PEC behavior of ZnIn2S4@NH2-MIL-125(Ti) composite could be attributed to that the Ti4+-Ti3+ intervalence cycles in the titanium oxo-cluster of NH2-MIL-125(Ti) as well as the matching energy level between ZnIn2S4 and NH2-MIL-125(Ti) promote the migration and separation of photocarrier. Besides, polydopamine (PDA) with abundant amino- and quinone-groups was selected to further improve the PEC signals and capture antibody, which implement through the covalent bonding of PDA and BSA-E2 or carboxyl-group functionalized Mn:ZnCdS QDs in the competitive-type strategy. Concretely, the quinone functional groups in PDA film was applied to immobilize BSA-E2 through Michael reactions, and the PDA nanosphere loaded Mn:ZnCdS quantum dot (PDA NS/Mn:ZnCdS QDs) was used as antibodies' labels to amplify PEC signals. After PDA NS/Mn:ZnCdS-anti-E2 immobilized on the modified electrode, a remarkable increase of photocurrent signal was observed owing to the specific bonding of antigen and antibody. Based on the competitive binding of PDA NS/Mn:ZnCdS-anti-E2 with either free E2 or bovine serum albumin (BSA)-E2 causing the change of the photocurrent signal, the standard sample free E2 could be accuracy detect. Under optimal conditions, the competitive-type PEC immunosensor exhibited the linear range from 0.0005â¯ng/mL to 20â¯ng/mL and a limit detection of 0.3â¯pg/mL (S/Nâ¯=â¯3). Meanwhile, the acceptable stability, selectivity and reproducibility of the proposed PEC immunosensing platform indicating the promising detection of small molecular environmental pollutants.
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Técnicas Biossensoriais/métodos , Estradiol/análise , Indóis/química , Nanocompostos/química , Polímeros/química , Titânio/química , Poluentes Químicos da Água/análise , Anticorpos Imobilizados/química , Compostos de Cádmio/química , Técnicas Eletroquímicas/métodos , Imunoensaio/métodos , Limite de Detecção , Dietilamida do Ácido Lisérgico/análogos & derivados , Dietilamida do Ácido Lisérgico/química , Nanocompostos/ultraestrutura , Pontos Quânticos/química , Pontos Quânticos/ultraestrutura , Sulfetos/química , Água/análise , Zinco/químicaRESUMO
A competitive immunosensor was established using an electrochemical amperometric strategy for sensitive detection of tetrabromobisphenol A bis(2-hydroxyethyl) ether (TBBPA-DHEE), an important derivative of tetrabromobisphenol A (TBBPA). In this system, the amplified electrochemical signal towards the reduction of hydrogen peroxide (H2O2) was recorded by amperometric method. Meanwhile, the synthetized catalase functionalized AuNPs-loaded self-assembled polymer nanospheres showed an excellent electrocatalytic ability to catalyse H2O2, which was beneficial for strengthening the electrochemical signals. Under the optimized conditions, this method displayed: (i) low detection limits (0.12â¯ng/mL, 7 times lower than the traditional ELISA with the same antibody); (ii) satisfactory accuracy (recoveries, 78-124%; RSD, 2.1-8.3%) and good agreement with the corresponding ELISA; (iii) low sample consumption (6⯵L) and low cost. The proposed approach was applied for investigation of TBBPA-DHEE from environmental waters, and our results indicated that this immunosensor has great potential to detect the trace pollutants in aquatic environments.
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Compostos Benzidrílicos/análise , Técnicas Eletroquímicas/métodos , Éteres/análise , Retardadores de Chama/análise , Hidrocarbonetos Bromados/análise , Imunoensaio/métodos , Fenóis/análise , Anticorpos Imobilizados/imunologia , Compostos Benzidrílicos/imunologia , Catalase/química , Éteres/imunologia , Água Doce/análise , Ouro/química , Hidrocarbonetos Bromados/imunologia , Peróxido de Hidrogênio/química , Limite de Detecção , Nanopartículas Metálicas/química , Fenóis/imunologia , Poluentes Químicos da Água/análiseRESUMO
A novel, rapid and convenient competitive immunoassay for ultrasensitive detection of chloramphenicol residues in shrimp and honey was established combined with flow injection chemiluminescence. The carboxylic resin beads were used as solid phase carriers to load with more coating antigen due to their larger specific surface area and good biocompatibility. The surface of the silica dioxide nanoparticles was modified with aldehyde group to combine with more horseradish peroxidase and the chloramphenicol antibody. There was a competitive process between the chloramphenicol in solution and the immobilized coating antigen to combine with the limited binding site of antibody to form the immunocomplex. Silica dioxide nanoparticles played an important role in enhancing chemiluminescence signal, because the horseradish peroxidase on SiO2 effectively catalyzed the system of luminol-PIP-H2O2. Under optimal conditions, the chemiluminescence intensity decreased linearly with the logarithm of the chloramphenicol concentration in the range of 0.0001 to 100â¯ngâ¯mL-1 and the detection limit (3σ) was 0.033â¯pgâ¯mL-1. This immunosensor demonstrated acceptable stability, high specificity and reproducibility. The horseradish peroxidase-silica dioxide nanoparticle-chloramphenicol antibody complex successfully prepared in this article was firstly applied to the detection of chloramphenicol, and had extremely important meanings for the application of nanoparticles and enzymatic catalysis in the field of chemiluminescence.
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Antibacterianos/análise , Cloranfenicol/análise , Análise de Alimentos/instrumentação , Medições Luminescentes/instrumentação , Nanopartículas/química , Dióxido de Silício/química , Animais , Anticorpos Monoclonais/química , Técnicas Biossensoriais/economia , Técnicas Biossensoriais/instrumentação , Desenho de Equipamento , Análise de Injeção de Fluxo/economia , Análise de Injeção de Fluxo/instrumentação , Análise de Alimentos/economia , Mel/análise , Imunoensaio/economia , Imunoensaio/instrumentação , Limite de Detecção , Medições Luminescentes/economia , Penaeidae/química , Reprodutibilidade dos Testes , Resinas Sintéticas/química , Frutos do Mar/análiseRESUMO
Since p53 protein has become recognized biomarker for both diagnostic and therapeutic purposes in oncological diseases with particular relevance for bladder cancer, it is highly desirable to search for a novel sensing tool for detecting the patient's p53 level at the early stage. Here we report the first study on the development and validation of a novel disposable competitive amperometric immunosensor for determination of p53 protein at subnanomolar levels, based on p53 immobilization on gold nanoparticles/carbon nanotubes modified screen-printed carbon electrodes. The assay protocol requires the use of single anti-p53 mouse monoclonal antibody (DO-7 clone), able to recognize both wild-type and mutant p53. The developed immunosensor as well as the protocol of the electrochemical immunoassay were optimized by means of an experimental design procedure to assess the suitability of the device to be validated and applied for the determination of p53 in untreated and undiluted urine samples. It was found that the developed competitive immunodevice was able to achieve wide linear range detection of wild-type p53 from 20 pM to 10 nM with a low detection limit of 14 pM in synthetic urine samples, suggesting the sensor's capability of working in a complex sample matrix. The excellent performance results also in terms of selectivity, trueness and precision, coupled with the advantages of an easy preparation and low-cost assay in contrast to other methods which require very complex, time-consuming and costly nanostructured architectures, makes the developed competitive immunosensor an analytically robust diagnostic tool, valuable for implementation of screening and follow-up programs in patients with urologic malignancies.