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A new type of label-free electrochemical immunosensor for the high-sensitivity determination of parathion was developed based on the oriented immobilization of nanobody (VHH9) on a gold nanoparticle-loaded polyvinyl alcohol/citric acid nanofiber membrane-modified electrode. The morphology characterization and assembly process of the modified materials were investigated using scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). Under the optimum conditions, the label-free electrochemical immunosensor for parathion exhibited a linear range of 0.0015-6400 ng/mL and a low detection limit of 0.48 pg/mL, the signal response of which was 10 times higher than that of the randomly immobilized VHH9. The immunosensor possessed high selectivity, good repeatability and reusability (keeping above 90% of its initial activity after repeating 8 times), and stability (remaining 90% after 9 weeks of storage). Finally, the average recoveries of parathion from food samples were 93.76-105.73% with the coefficient of variation being 2.65-6.85%, showing good correlation with UPLC (R2 = 0.9950). Therefore, our nanobody immobilization protocol is simple and effective and proves the potential to be utilized as a promising candidate for sensing platform.
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Técnicas Biossensoriais , Técnicas Eletroquímicas , Ouro , Limite de Detecção , Nanofibras , Nanofibras/química , Ouro/química , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Anticorpos de Domínio Único/imunologia , Anticorpos de Domínio Único/química , Nanopartículas Metálicas/química , Imunoensaio/métodos , Anticorpos Imobilizados/imunologia , Eletrodos , Álcool de Polivinil/química , Contaminação de Alimentos/análise , Membranas ArtificiaisRESUMO
Artificial spin ices are engineered arrays of dipolarly coupled nanobar magnets. They enable direct investigations of fascinating collective phenomena from their diverse microstates. However, experimental access to ground states in the geometrically frustrated systems has proven difficult, limiting studies and applications of novel properties and functionalities from the low energy states. Here, we introduce a convenient approach to control the competing diploar interactions between the neighboring nanomagnets, allowing us to tailor the vertex degeneracy of the ground states. We achieve this by tuning the length of selected nanobar magnets in the spin ice lattice. We demonstrate the effectiveness of our method by realizing multiple low energy microstates in a kagome artificial spin ice, particularly the hardly accessible long range ordered ground state-the spin crystal state. Our strategy can be directly applied to other artificial spin systems to achieve exotic phases and explore new emergent collective behaviors.
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KEY MESSAGE: A quantitative trait locus GW10 is located on Chromosome 10 by map-based cloning, which encodes a P450 Subfamily protein. The GW10 regulates grain size and grain number in rice involved in the BR pathway. Grain size and grain number play extremely important roles in rice grain yield. Here, we identify GW10, which encodes a P450 subfamily protein and controls grain size and grain number by using Lemont (tropical japonica) as donor parent and HJX74 (indica) as recipient parent. The GW10 locus was mapped into a 14.6 kb region in HJX74 genomic on the long arm of chromosome 10. Lower expression of the gw10 in panicle is contributed to the shorter and narrower rice grain, and the increased number of grains per panicle. In contrast, overexpression of GW10 is contributed to longer and wider rice grain. Furthermore, the higher expression levels of some of the brassinosteroid (BR) biosynthesis and response genes are associated with the NIL-GW10. The sensitivity of the leaf angle to exogenous BR in NIL-GW10 is lower than that in NIL-gw10 and in the KO-GW10, which implied that the GW10 should involve in the brassinosteroid-mediated regulation of rice grain size and grain number.
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Sistema Enzimático do Citocromo P-450/genética , Oryza/genética , Locos de Características Quantitativas , Sementes/crescimento & desenvolvimento , Mapeamento Cromossômico , Cruzamentos Genéticos , Grão Comestível/genética , Genes de Plantas , Oryza/crescimento & desenvolvimentoRESUMO
The ability to control the potential landscape in a medium of interacting particles could lead to intriguing collective behavior and innovative functionalities. Here, we utilize spatially reconfigurable magnetic potentials of a pinwheel artificial-spin-ice (ASI) structure to tailor the motion of superconducting vortices. The reconstituted chain structures of the magnetic charges in the pinwheel ASI and the strong interaction between magnetic charges and superconducting vortices allow significant modification of the transport properties of the underlying superconducting thin film, resulting in a reprogrammable resistance state that enables a reversible and switchable vortex Hall effect. Our results highlight an effective and simple method of using ASI as an in situ reconfigurable nanoscale energy landscape to design reprogrammable superconducting electronics, which could also be applied to the in situ control of properties and functionalities in other magnetic particle systems, such as magnetic skyrmions.
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The motion of Abrikosov vortices in type-II superconductors results in a finite resistance in the presence of an applied electric current. Elimination or reduction of the resistance via immobilization of vortices is the "holy grail" of superconductivity research. Common wisdom dictates that an increase in the magnetic field escalates the loss of energy since the number of vortices increases. Here we show that this is no longer true if the magnetic field and the current are applied parallel to each other. Our experimental studies on the resistive behavior of a superconducting Mo0.79Ge0.21 nanostrip reveal the emergence of a dissipative state with increasing magnetic field, followed by a pronounced resistance drop, signifying a reentrance to the superconducting state. Large-scale simulations of the 3D time-dependent Ginzburg-Landau model indicate that the intermediate resistive state is due to an unwinding of twisted vortices. When the magnetic field increases, this instability is suppressed due to a better accommodation of the vortex lattice to the pinning configuration. Our findings show that magnetic field and geometrical confinement can suppress the dissipation induced by vortex motion and thus radically improve the performance of superconducting materials.
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Tetrabromobisphenol A (TBBPA) is the largest brominated flame retardant which can be released to environment and cause long-term hazard. In this work, we developed a rapid and highly sensitive fluorescence enzyme-linked immunosorbent assay (FELISA) for monitoring of TBBPA in soil samples. TBBPA specific nanobody derived from camelid was fused with alkaline phosphatase to obtain the bi-functional fusion protein, which enable the specific binding of TBBPA and the generation of detection signal simultaneously. The assay showed an IC50 of 0.23â¯ngâ¯g-1, limit detection of 0.05â¯ngâ¯g-1 and linear range from 0.1 to 0.55â¯ngâ¯g-1 for TBBPA in soil samples. Due to the high resistance to organic solvents of the fusion protein, a simple pre-treatment by using 40% dimethyl sulfoxide (DMSO) as extract solvent can eliminate matrix effect and obtain good recoveries (ranging from 93.4% to 112.4%) for spiked soil samples. Good relationship between the results of the proposed FELISA and that of liquid chromatography tandem mass spectrometry (LC-MS/MS) was obtained, which indicated it could be a powerful analytical tool for determination of TBBPA to monitor human and environmental exposure.
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Monitoramento Ambiental/métodos , Ensaio de Imunoadsorção Enzimática , Retardadores de Chama/análise , Bifenil Polibromatos/análise , Poluentes do Solo/análise , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Camelídeos Americanos , Limite de Detecção , Bifenil Polibromatos/metabolismo , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Anticorpos de Domínio Único/genética , Anticorpos de Domínio Único/metabolismoRESUMO
1-naphthol (1-NAP) is the main metabolite of pesticide carbaryl and naphthalene, and is also a genotoxic and carcinogenic intermediate in the synthesis of organic compound, dyes, pigment and pharmaceutical industry. In this work, two novel haptens were designed and synthesized for developing a competitive indirect enzyme-linked immunosorbent assay (ciELISA) method for 1-NAP in urine samples. The assay showed a limit of detection of 2.21 ng/mL and working range from 4.02 ng/mL to 31.25 ng/mL for 1-NAP in optimized working buffer. The matrix effect of samples was eliminated via 15-fold dilution of optimized working buffer. Good average recoveries (102.4%-123.4%) with a coefficient of variation from 11.7% to 14.7% was obtained for spiked urine samples. Subsequent instrument verification test showed good correlation between the results of ciELISA and high-performance liquid chromatography. The developed ciELISA is a high-throughput tool to monitor 1-NAP in urine, which can provide technical support for the establishment of biological exposure level for the exposure to carbaryl, naphthalene and other related pollutants.
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Anticorpos Monoclonais/química , Ensaio de Imunoadsorção Enzimática/métodos , Haptenos/química , Naftóis/urina , Resíduos de Praguicidas/urina , Anticorpos Monoclonais/imunologia , Carbaril/metabolismo , Exposição Ambiental/análise , Limite de Detecção , Naftalenos/metabolismo , Naftóis/imunologia , Resíduos de Praguicidas/imunologia , Resíduos de Praguicidas/metabolismoRESUMO
We demonstrate strong magnon-photon coupling of a thin-film Permalloy device fabricated on a coplanar superconducting resonator. A coupling strength of 0.152 GHz and a cooperativity of 68 are found for a 30-nm-thick Permalloy stripe. The coupling strength is tunable by rotating the biasing magnetic field or changing the volume of Permalloy. We also observe an enhancement of magnon-photon coupling in the nonlinear regime of the superconducting resonator, which is attributed to the nucleation of dynamic flux vortices. Our results demonstrate a critical step towards future integrated hybrid systems for quantum magnonics and on-chip coherent information transfer.
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BACKGROUND: The aim of this study is to evaluate the effects of recombinant human bone morphogenetic protein 2 (rhBMP-2), human amnion mesenchymal stem cells (hAMSCs), and nanohydroxyapatite/collagen/poly(l-lactide) (nHAC/PLA) in tissue engineering to provide potential approaches for periodontal bone regeneration. METHODS: hAMSCs were isolated from discarded amniotic membrane samples and cultured in vitro. Alkaline phosphatase (ALP) staining and alizarin red staining were performed to evaluate the osteoblast (OB) differentiation ability of hAMSCs. Three groups were divided: the experimental group (cells transfected with pcDNA3.1-rhBMP-2), the blank group (cells without gene transfection), and the control group (cells transfected with empty plasmid). RT-PCR and western blot were used to examine whether rhBMP-2 has been successfully expressed. 3-(4,5)-dimethylthiahiazol(-z-y1)-3,5-di-phenytetrazo-liumromide assay (MTT) was done to detect the effect of rhBMP-2 on hAMSCs seeded on nHAC/PLA. ALP activity, mineral formation assay, calcium, phosphate and osteocalcin (OCN) content, and OCN and RUNX2 expression of hAMSCs were detected to evaluate osteogenic differentiation capability of rhBMP-2 on hAMSCs seeded on nHAC/PLA. RESULTS: hAMSCs exhibited intense ALP staining, obvious calcium deposition, and mineralization nodules, and rhBMP-2 were highly expressed in the experimental group. The proliferation of the hAMSCs with rhBMP-2 on nHAC/PLA was significantly higher than the cells without rhBMP-2, and the cells all increased in a time-dependent manner. rhBMP-2 significantly increased the OCN and phosphate content, mineral formation, ALP activity, osteogenic biomarkers OCN, and Runx2, and decreased calcium content in hAMSCs seeded on the nHAC/PLA scaffold. CONCLUSIONS: This finding demonstrated that hAMSCs has an ideal OB differentiation ability. rhBMP-2 facilitates the proliferation and osteogenesis of hAMSCs. The nHAC/PLA could act as a good scaffold for hAMSCs seeding, proliferation, and osteogenic differentiation. The application of rhBMP-2, nHAC/PLA, and hAMSCs in tissue engineering may offer promising possibilities for periodontal bone regeneration.
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Âmnio/citologia , Âmnio/efeitos dos fármacos , Proteína Morfogenética Óssea 2/farmacologia , Diferenciação Celular/efeitos dos fármacos , Colágeno/química , Durapatita/química , Células-Tronco Mesenquimais/citologia , Osteoblastos/citologia , Poliésteres/química , Proteína Morfogenética Óssea 2/genética , Regeneração Óssea/efeitos dos fármacos , Cálcio/metabolismo , Proliferação de Células/efeitos dos fármacos , Humanos , Técnicas In Vitro , Células-Tronco Mesenquimais/química , Osteoblastos/química , Osteocalcina/metabolismo , Osteogênese/efeitos dos fármacos , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Engenharia TecidualRESUMO
Electronic textiles (E-textiles) offer great wearing comfort and unobtrusiveness, thus holding potential for next-generation health monitoring wearables. However, the practical implementation is hampered by challenges associated with poor signal quality, substantial motion artifacts, durability for long-term usage, and non-ideal user experience. Here, we report a cost-effective E-textile system that features 3D microfiber-based electrodes for greatly increasing the surface area. The soft and fluffy conductive microfibers disperse freely and securely adhere to the skin, achieving a low impedance at the electrode-skin interface even in the absence of gel. A superhydrophobic fluorinated self-assembled monolayer was deposited on the E-textile surface to render it waterproof while retaining the electrical conductivity. Equipped with a custom-designed motion-artifact canceling wireless data recording circuit, the E-textile system could be integrated into a variety of smart garments for exercise physiology and health monitoring applications. Real-time multimodal electrophysiological signal monitoring, including electrocardiogram (ECG) and electromyography (EMG), was successfully carried out during strenuous cycling and even underwater swimming activities. Furthermore, a multi-channel E-textile was developed and implemented in clinical patient studies for simultaneous real-time monitoring of maternal ECG and uterine EMG signals, incorporating spatial-temporal potential mapping capabilities.
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N6-methyladenosine (m6A) is a prevalent internal post-transcriptional modification in eukaryotic RNAs executed by m6A-binding proteins known as "readers." Our previous research demonstrated that the Arabidopsis m6A reader ECT2 positively regulates transcript levels of the proteasome regulator PTRE1 and several 20S proteasome subunits, thereby enhancing 26S proteasome activity. However, mechanism underlying the selective recognition of m6A targets by readers, such as ECT2, remains elusive. In this study, we further demonstrate that ECT2 physically interacts with PTRE1 and several 20S proteasome subunits. This interaction, which occurs on the ribosome, involves the N terminus of PTRE1, suggesting that ECT2 might bind to the nascent PTRE1 polypeptide. Deleting ECT2's protein interaction domain impairs its mRNA-binding ability, whereas mutations in the m6A-RNA-binding site do not affect protein-protein interactions. Moreover, introducing a novel protein-binding domain into ECT2 increases transcript levels of proteins interacting with this domain. Our findings indicate that interaction with the PTRE1 protein enhances ECT2's binding to PTRE1 m6A mRNAs during translation, thereby regulating PTRE1 mRNA levels.
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Emerging from the intricate interplay of topology and magnetism, the giant anomalous Hall effect (AHE) is the most known topological property of the recently discovered kagomé ferromagnetic Weyl semimetal Co3Sn2S2 with the magnetic Co atoms arranged on a kagomé lattice. Here it is reported that the AHE in Co3Sn2S2 can be fine-tuned by an applied magnetic field orientated within ≈2° of the kagomé plane, while beyond this regime, it stays unchanged. Particularly, it can vanish in magnetic fields parallel to the kagomé plane and even decrease in magnetic fields collinear with the spin direction. This tunable AHE can be attributed to local spin switching enabled by the geometrical frustration of the magnetic kagomé lattice, revealing that spins in a kagomé ferromagnet change their switching behavior as the magnetic field approaches the kagomé plane. These results also suggest a versatile way to tune the properties of a kagomé magnet.
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With the popularity of herbal tea in China, many food fraudsters have added illegal drugs to herbal tea to enhance its functions, among which aminopyrine is widely abused as an antipyretic and analgesic. Presently, there is no immunoassays for aminopyrine, and it is difficult to achieve real-time detection in the field. Based on a polyclonal antibody of aminopyrine with high specificity and sensitivity, an optimal combination of coating antigen/antibody was obtained by screening different coating antigens. On this basis, a sensitive ic-ELISA method was established to detect aminopyrine in herbal tea. The detection limit of the ic-ELISA was 0.18 ng mL-1, which was much lower than the 100 ng mL-1 required as a standard. The method had good consistency with LC-MS in the detection of actual samples and could be used as a reliable method for the detection of aminopyrine in herbal tea.
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Chás de Ervas , Aminopirina , Imunoensaio/métodos , Ensaio de Imunoadsorção Enzimática/métodos , AnticorposRESUMO
Porcine epidemic diarrhea (PED) is a severe contagious intestinal disease caused by the porcine epidemic diarrhea virus (PEDV), which leads to high mortality in piglets. In this study, by analyzing a total of 53 full-length spike genes and COE domain regions of PEDVs, the conserved COE fragment of the spike protein from the dominant strain SC1402 was chosen as the target protein and expressed successfully in Pichia pastoris (P. pastoris). Furthermore, an indirect enzyme-linked immunosorbent assay (iELISA) based on the recombinant COE protein was developed for the detection of anti-PEDV antibodies in pig sera. The results showed that under the optimized conditions, the cut-off value of COE-based indirect ELISA (COE-iELISA) was determined to be 0.12. Taking the serum neutralization test as standard, the relative sensitivity of the COE-iELISA was 94.4% and specificity 92.6%. Meanwhile, no cross-reactivity to other porcine pathogens was noted with this assay. The intra-assay and inter-assay coefficients of variation were less than 7%. Moreover, 164 vaccinated serum samples test showed that overall agreement between COE-iELISA and the actual diagnosis result was up to 99.4%. More importantly, the developed iELISA exhibited a 95.08% agreement rate with the commercial ELISA kit (Kappa value = 0.88), which suggested that the expressed COE protein was an effective antigen in serologic tests and the established COE-iELISA is reliable for monitoring PEDV infection in pigs or vaccine effectiveness.
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Infecções por Coronavirus , Vírus da Diarreia Epidêmica Suína , Doenças dos Suínos , Animais , Suínos , Epitopos , Vírus da Diarreia Epidêmica Suína/genética , Saccharomyces cerevisiae , Anticorpos Antivirais , Ensaio de Imunoadsorção Enzimática/métodos , Proteínas Recombinantes/genética , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/prevenção & controleRESUMO
Electromyometrial imaging (EMMI) technology has emerged as one of the promising technology that can be used for non-invasive pregnancy risk stratification and for preventing complications due to pre-term birth. Current EMMI systems are bulky and require a tethered connection to desktop instrumentation, as a result, the system cannot be used in non-clinical and ambulatory settings. In this article, we propose an approach for designing a scalable, portable wireless EMMI recording system that can be used for in-home and remote monitoring. The wearable system uses a non-equilibrium differential electrode multiplexing approach to enhance signal acquisition bandwidth and to reduce the artifacts due to electrode drifts, amplifier 1/f noise, and bio-potential amplifier saturation. A combination of active shielding, a passive filter network, and a high-end instrumentation amplifier ensures sufficient input dynamic range ([Formula: see text]) such that the system can simultaneously acquire different bio-potential signals like maternal electrocardiogram (ECG) in addition to the EMMI electromyogram (EMG) signals. We show that the switching artifacts and the channel cross-talk introduced due to non-equilibrium sampling can be reduced using a compensation technique. This enables the system to be potentially scaled to a large number of channels without significantly increasing the system power dissipation. We demonstrate the feasibility of the proposed approach in a clinical setting using an 8-channel battery-powered prototype which dissipates less than 8 µW per channel for a signal bandwidth of 1 KHz.
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Processamento de Sinais Assistido por Computador , Dispositivos Eletrônicos Vestíveis , Eletrocardiografia , Eletromiografia , Eletrodos , Tecnologia sem FioRESUMO
Titanium nitride is a material of interest for many superconducting devices such as nanowire microwave resonators and photon detectors. Thus, controlling the growth of TiN thin films with desirable properties is of high importance. This work aims to explore effects in ion beam-assisted sputtering (IBAS), were an observed increase in nominal critical temperature and upper critical fields are in tandem with previous work on Niobium nitride (NbN). We grow thin films of titanium nitride by both, the conventional method of DC reactive magnetron sputtering and the IBAS method, to compare their superconducting critical temperatures [Formula: see text] as functions of thickness, sheet resistance, and nitrogen flow rate. We perform electrical and structural characterizations by electric transport and x-ray diffraction measurements. Compared to the conventional method of reactive sputtering, the IBAS technique has demonstrated a 10% increase in nominal critical temperature without noticeable variation in the lattice structure. Additionally, we explore the behavior of superconducting [Formula: see text] in ultra-thin films. Trends in films grown at high nitrogen concentrations follow predictions of mean-field theory in disordered films and show suppression of superconducting [Formula: see text] due to geometric effects, while nitride films grown at low nitrogen concentrations strongly deviate from the theoretical models.
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For monitoring of the residual of parathion pesticide in food, herein, a sensitive and reliable electrochemical immunosensor based on cross-linked polyvinyl alcohol/citric acid nanofiber membrane (PVA/CA NFM) and horseradish peroxidase labeled anti-parathion nanobody was constructed. Firstly, the cross-linked PVA/CA NFM with extra-high surface area and uniform morphology was prepared and characterized. Then, the immunosensor was assembled and its analytical performances were evaluated. It exhibited high specificity and sensitivity to parathion with the liner range and limit of detection being 0.01-100 ng/mL and 2.26 pg/mL, respectively. Moreover, the biosensor kept almost 75% of its initial activity after regenerating 4 times, and remained 85% after 9 weeks of storage. Finally, the average recoveries from food samples were 96.20%-114.61% with the coefficient of variation being 1.06%-5.28%, which was correlate well with UPLC (R2 = 0.9964). Therefore, the sensor was demonstrated to be a feasible alternative for sensitive assay of parathion.
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Técnicas Biossensoriais , Nanocompostos , Paration , Técnicas Eletroquímicas , Limite de Detecção , Imunoensaio , Nanocompostos/química , Álcool de Polivinil/química , Ouro/químicaRESUMO
Paclobutrazol (PBZ) is a plant growth inhibitor and fungicide, but it is also carcinogenic and teratogenic, and has potential harm to human health. In this study, two PBZ haptens (PBZ-1, PBZ-2) were synthesized and conjugated with carrier proteins to get artificial antigens. A highly specific monoclonal antibody (mAb) against PBZ was prepared. The antibody subtype was IgG1 and the concentration was 11.03 mg/mL. A sensitive and rapid time-resolved fluorescence microsphere lateral flow immunoassay (TRFMs-LFIA) was established based on the mAb. The activated pH, the mAbs diluents, the mAb reacting concentration and the probe amount were optimized. The visual limit of detection (vLOD) and quantitative limit of detection (qLOD) of the TRFMs-LFIA for PBZ were 50 and 1.72 ng/mL respectively, and the 50% inhibiting concentration (IC50) was 9.38 ng/mL. The pretreatment procedures are simple and rapid, and the detection time of TRFMs-LFIA strip is 6 min. Qualitative and quantitative analysis of PBZ could be achieved under a UV light or with a portable fluorescence immunoassay analyzer. The average recovery rates ranged from 96.2% to 111.9% and the corresponding coefficients of variation (CV) were 4.0%-11.2% in spiked wheat and rice samples. Twenty real wheat and rice samples were measured by the TRFMs-LFIA and compared with Ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS). The measured values showed a good accordance. These results indicated that the proposed assay will provide a novel effective strategy for on-site detection of PBZ.
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In this study, forchlorfenuron (CPPU) was coupled with succinic anhydride to yield a CPPU hapten (CPPU-COOH), and a high-affinity monoclonal antibody (mAb) that can specifically recognize CPPU was produced. Using this mAb as a recognition reagent, a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) for CPPU was optimized, which exhibits an IC50 of 1.04 ng/mL, a limit of detection of 0.16 ng/mL, and a linear range of 0.31-3.43 ng/mL for CPPU. Cross-reactivity percentages with six analogues were all below 6%. The average recovery rates for cucumber and orange samples were from 85.23% to 119.14%. The analysis results of this icELISA showed good consistency with those from liquid chromatography mass spectrometry. These results suggest that the proposed icELISA provides a sensitive, specific, and reliable strategy for CPPU detection in food samples.
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Anticorpos Monoclonais , Compostos de Fenilureia , Anticorpos Monoclonais/análise , Ensaio de Imunoadsorção Enzimática/métodos , PiridinasRESUMO
A novel and efficient immunoaffinity column (IAC) based on bispecific monoclonal antibody (BsMAb) recognizing aflatoxin B1 (AFB1) and ochratoxin A (OTA) was prepared and applied in simultaneous extraction of AFB1 and OTA from food samples and detection of AFB1/OTA combined with ic-ELISA (indirect competitive ELISA). Two deficient cell lines, hypoxanthine guanine phosphoribosyl-transferase (HGPRT) deficient anti-AFB1 hybridoma cell line and thymidine kinase (TK) deficient anti-OTA hybridoma cell line, were fused to generate a hybrid-hybridoma producing BsMAb against AFB1 and OTA. The subtype of the BsMAb was IgG1 via mouse antibody isotyping kit test. The purity and molecular weight of BsMAb were confirmed by SDS-PAGE method. The cross-reaction rate with AFB2 was 37%, with AFG1 15%, with AFM1 48%, with AFM2 10%, and with OTB 36%. Negligible cross-reaction was observed with other tested compounds. The affinity constant (Ka) was determined by ELISA. The Ka (AFB1) and Ka (OTA) was 2.43 × 108 L/mol and 1.57 × 108 L/mol, respectively. Then the anti-AFB1/OTA BsMAb was coupled with CNBr-Sepharose, and an AFB1/OTA IAC was prepared. The coupling time and elution conditions of IAC were optimized. The coupling time was 1 h with 90% coupling rate, the eluent was methanol-water (60:40, v:v, pH 2.3) containing 1 mol/L NaCl, and the eluent volume was 4 mL. The column capacities of AFB1 and OTA were 165.0 ng and 171.3 ng, respectively. After seven times of repeated use, the preservation rates of column capacity for AFB1 and OTA were 69.3% and 68.0%, respectively. The ic-ELISA for AFB1 and OTA were applied combined with IAC. The IC50 (50% inhibiting concentration) of AFB1 was 0.027 ng/mL, the limit of detection (LOD) was 0.004 ng/mL (0.032 µg/kg), and the linear range was 0.006 ng/mL~0.119 ng/mL. The IC50 of OTA was 0.878 ng/mL, the LOD was 0.126 ng/mL (1.008 µg/kg), and the linear range was 0.259 ng/mL~6.178 ng/mL. Under optimum conditions, corn and wheat samples were pretreated with AFB1-OTA IAC. The recovery rates of AFB1 and OTA were 95.4%~105.0% with ic-ELISA, and the correlations between the detection results and LC-MS were above 0.9. The developed IAC combined with ic-ELISA is reliable and could be applied to the detection of AFB1 and OTA in grains.