A novel and ultrasensitive sandwich-type electrochemical aptasensor has been developed for the detection of thrombin, based on dual signal-amplification using HRP and apoferritin. Core/shell Fe3O4/Au magnetic nanoparticles (AuMNPs) loading aptamer1 (Apt1) was used as recognition elements, and apoferritin dually labeled with Aptamer2 (Apt2) and HRP was used as a detection probe. Sandwich-type complex, Apt1/thrombin/Apt2–apoferritin NPs–HRP was formed by the affinity reactions between AuMNPs–Apt1, thrombin, and Apt2–apoferritin–HRP. The complex was anchored on a screen-printed carbon electrode (SPCE). Differential pulse voltammetry (DPV) was used to monitor the electrode response. The proposed aptasensor yielded a linear current response to thrombin concentrations over a broad range of 0.5–100 pM with a detection limit of 0.07 pM (S/N = 3). The detection signal was amplified by using apoferritin and HRP. This nanoparticle-based aptasensor offers a new method for rapid, sensitive, selective, and inexpensive quantification of thrombin, and offers a promising potential in protein detection and disease diagnosis. 相似文献
We propose a separation/concentration‐signal‐amplification in‐one method based on electrochemical conversion (ECC) of magnetic nanoparticles (MNPs) to develop a facile and sensitive electrochemical biosensor for chloramphenicol (CAP) detection. Briefly, aptamer‐modified magnetic nanoparticles (MNPs‐Apt) was designed to capture CAP in sample, then the MNPs‐Apt composite was conjugated to Au electrode through the DNA hybridization between the unoccupied aptamer and a strand of complementary DNA. The ECC method was applied to transfer MNPs labels to electrochemically active Prussian blue (PB). The anodic and cathodic currents of PB were taken for signal readout. Comparing with conventional methods that require electrochemically active labels and related sophisticated labelling procedures, this method explored and integrated the magnetic and electrochemical properties of MNPs into one system, in turn realized magnetic capturing of CAP and signal generation without any additional conventional labels. Taking advantages of the high abundance of iron content in MNPs and the refreshing effect deriving from ECC process, the method significantly promoted the signal amplification. Therefore, the proposed biosensors exhibited linear detection range from 1 to 1000 ng mL−1 and a limit of detection down to 1 ng mL−1, which was better than or comparable with those of most analogues, as well as satisfactory specificity, storage stability and feasibility for real samples. The developed method may lead to new concept for rapid and facile biosensing in food safety, clinic diagnose/therapy and environmental monitoring fields. 相似文献
A simple and label-free electrochemical aptasensor was developed for ultra-sensitive determination of chloramphenicol (CAP) based on a 2D transition of metal carbides (MXene) loaded with gold nanoparticles (AuNPs). The embedded AuNPs not only inhibit the aggregation of MXene sheets, but also improve the quantity of active sites and electronic conductivity. The aptamers (Apts) were able to immobilize on the MXene–AuNP modified electrode surface through Au–S interaction. Upon specifically binding with CAP with high affinity, the CAP–Apt complexes produced low conductivity on the aptasensor surface, leading to a decreased electrochemical signal. The resulting current change was quantitatively correlated with CAP concentration. Under optimized experimental conditions, the constructed aptasensor exhibited a good linear relationship within a wide range of 0.0001–10 nM and with a low detection limit of 0.03 pM for CAP. Moreover, the developed aptasensor has been applied to the determination of CAP concentration in honey samples with satisfactory results. 相似文献
The authors report on an aptamer-based electrochemical assay for the insecticide acetamiprid. It is based on the target-induced release of the redox probe Methylene Blue (MB) from the dsDNA formed between aptamer and complementary strand (Apt/CS), exonuclease I (Exo I) and silica nanoparticles coated with streptavidin (SiNP-Streptavidin). MB is detected electrochemically using an unmodified gold electrode. In the presence of acetamiprid, MB is released from the Apt/CS dsDNA and accumulated in the close environment of the gold electrode. This results in a strong electrochemical signal for MB at fairly low working voltage of typically ?0.27 V. In the absence of target, however, the SiNP-streptavidin conjugate modified MB-dsDNA remains intact. Hence, the electrochemical signal remains weak. The method displays high selectivity for acetamiprid and a limit of detection as low as 153 pM. The assay was successfully applied to the determination of acetamiprid in (spiked) water and serum samples, with LODs of 161 and 209 pM, respectively.
Graphical abstract Schematic illustration of acetamiprid detection based on electrochemical assay. In the absence of acetamiprid, MB-dsDNA-modified SiNP-Streptavidin complex is intact and redox probe (MB) does not exist in the environment of electrode, resulting in a weak electrochemical signal (a). In the presence of target, Apt binds to acetamiprid and CS and MB leave the SiNP-Streptavidin. Exo I digests CS. So, a huge amount of MB is present in the environment of electrode and a strong electrochemical signal is observed (b).
In this paper, the design of a novel sandwich-type electrochemical aptasensor was reported for an ultrasensitive mercury ion (Hg2+) detection in water samples, which labeled with two-labeled aptamer (Apt) sequences. The used Apts were Apt1 and Apt2 as the capture and signal probe, respectively. The Apt1 probe was immobilized on the poly(4-aminobenzoic acid) (p-ABA) and quantum dots (QDs) film as the platform, as well as the Apt2 reporter was labeled with ferrocene. In the presence of Hg2+, the strong coordination complex has been formed between the specific thymine of the Apt1, Hg2+, as well as the thymine of the Apt as T–Hg2+–T adduct. The QDs and p-ABA were applied for increasing the conductivity of platform and suitable binding of the recognition elements. Under the optimized conditions, the constructed aptasensor illustrated either a wide linear relationship between the logarithm of Hg2+ concentration and current, from 0.05 to 100 nM and also an excellent low limit of detection of 0.01 nM. The quality of carefully choosing, an excellent stability and specificity sensitivity of the designed aptasensor, was investigated by spiked tap water samples as real sample. Moreover, the aptasensor exhibits the good reproducibility as well as has high selectivity for the other cations. The recoveries of the Hg2+ assay of the tap water samples were acquired satisfactorily which imply the generated aptasensor can use Hg2+ measurement in the real laboratories.
An electrochemical nanoaptasensor is described that is based on the use of a glassy carbon electrode (GCE) modified with electrodeposited silver nanoparticles (AgNPs). An aptamer (Apt) against trinitrotoluene (TNT) was then immobilized on the AgNPs. The addition of TNT to the modified GCE leads to decrease in peak current (typically measured at a potential of ?0.45 V vs. Ag/AgCl) of riboflavin which acts as an electrochemical probe. Even small changes in the surface (as induced by binding of Apt to TNT) alter the interfacial properties. As a result, the LOD is lowered to 33 aM, and the dynamic range extends from 0.1 fM to 10 μM without sacrificing specificity.
Graphical abstract Schematic presentation of a nanoaptasensor which is based on a glassy carbon electrode (GCE) modified with electrodeposited silver nanoparticles (AgNPs) and aptamer (Apt). It was applied to the detection of 2,4,6-trinitrotoluene (TNT) with the help of riboflavin (RF) as a redox probe.
A nanocomposite prepared from reduced graphene oxide (rGO) and silver nanoparticles (AgNPs) is used in an electrochemical aptasensor for the sensitive and selective determination of the antibiotic chloramphenicol (CAP). The nanocomposite was obtained by electrostatic assembly of AgNPs on the surface of polyelectrolyte-functionalized rGO and then used to modify a glassy carbon electrode. The biosensor is then obtained by immobilizing the aptamer against CAP. When incubated with solutions of CAP, the sensor surface is loaded with CAP due to aptamer recognition. The captured CAP can be electrochemically reduced to yield a current that is strongly enhanced as a result of the excellent electrocatalysis property of the graphene/AgNP-nanocomposite. Under optimum conditions, the calibration plot is linear in the 0.01 to 35 μM concentration range, with a 2 nM detection limit (at 3σ). The sensor is reproducible, stable, selective over homologous interferents, and performs excellently when analyzing CAP in milk samples.
Graphical Abstract A graphene/silver nanoparticle-based electrochemical aptasensor is designed for the selective determination of the antibiotic chloramphenicol (CAP). The excellent electrocatalytic reduction of CAP specifically captured onto the electrode surface enables the sensitive electrochemical signal transduction of the biosensor by linear sweep voltammetry (LSV).
We report on a protocol for a simultaneous competitive immunoassay for tetracycline (TC) and chloramphenicol (CAP) on the same sensing interface. Conjugates of TC and of CAP with bovine serum albumin were first co-immobilized on a glassy carbon electrode modified with gold nanoparticles. In parallel, monoclonal anti-TC and anti-CAP antibodies were conjugated onto CdS and PbS nanoclusters, respectively. In a typical assay, the immobilized haptens and the added target analytes competed for binding to the corresponding antibodies on the nanoclusters. Subsequently, Cd(II) and Pb(II) ions are released from the surface of the corresponding nanoclusters by treatment with acid and then were detected by square wave anodic stripping voltammetry. The currents at the peak potentials for Cd(II) and Pb(II) were used as the sensor signal for TC and CAP, respectively. This multiplex immunoassay enables the simultaneous determination of TC and CAP in a single run with dynamic ranges from 0.01 to 50 ng mL?1 for both analytes. The detection limits for TC and for CAP are 7.5 pg mL?1 and 5.4 pg mL?1, respectively. No obvious nonspecific adsorption and cross-reactivity was observed in a series of analyses. Intra-assay and inter-assay coefficients of variation were less than 10 %. The method was evaluated by analyzing TC and CAP in spiked samples of milk and honey. The recoveries range from 88 % to 107 % for TC, and from 91 % to 119 % for CAP.
Figure
We developed a new multiplexed electrochemical immunoassay for simultaneous determination of tetracycline and chloramphenicol, using metal sulfide nanoclusters as recognition elements. 相似文献
Electrochemical sensing has established a strong presence in diverse areas. The conventional electrochemical sensing approach consumes large sample volumes and reagents and requires bulky potentiostat, macro-electrodes, and other equipment. The synergistic integration of electrochemical sensing systems with miniaturized or microfluidic electrochemical devices and microelectrodes in a single platform provides rapid analysis with a disposable, reusable, and cost-effective platform for multiplexed point-of-care detections. Such microdevices have created scope for using several materials as electrodes and sensing platforms by using appropriate fabrication techniques. One of the most recent advancements in miniaturized devices includes the integration of automation and Internet of Things to realize fully automated and robust electrochemical microdevices. The review summarizes the emerging trends in fabrication methods of miniaturized and microfluidic devices, their multiple applications in real-time, integration of Internet of Things, automation, identifying research gaps with strategies for bridging these gaps, future outlook, and recent approaches to intelligent electrochemical sensing. 相似文献
A novel strategy for selective collection and detection of breast cancer cells (MCF-7) based on aptamer–cell interaction was developed. Mucin 1 protein (MUC1) aptamer (Apt1) was covalently conjugated to magnetic beads to capture MCF-7 cell through affinity interaction between Apt1 and MUC1 protein that overexpressed on the surface of MCF-7 cells. Meanwhile, a nano-bio-probe was constructed by coupling of nucleolin aptamer AS1411 (Apt2) to CdTe quantum dots (QDs) which were homogeneously coated on the surfaces of monodispersed silica nanoparticles (SiO2 NPs). The nano-bio-probe displayed similar optical and electrochemical performances to free CdTe QDs, and remained high affinity to nucleolin overexpressed cells through the interaction between AS1411 and nucleolin protein. Photoluminescence (PL) and square-wave voltammetric (SWV) assays were used to quantitatively detect MCF-7 cells. Improved selectivity was obtained by using these two aptamers together as recognition elements simultaneously, compared to using any single aptamer. Based on the signal amplification of QDs coated silica nanoparticles (QDs/SiO2), the detection sensitivity was enhanced and a detection limit of 201 and 85 cells mL−1 by PL and SWV method were achieved, respectively. The proposed strategy could be extended to detect other cells, and showed potential applications in cell imaging and drug delivery. 相似文献
Nanowires of MnO2 were prepared by a simple method in which the commercial granular -MnO2 powders were hydrothermally treated in water or ammonia solution at 150 °C. These 1D nanostructured manganese oxides were characterized physically by X-ray diffraction, scanning electron microscopy and transmission electron microscopy tests. Cyclic voltammetry and constant current discharge experiments were employed to explore the diversity of electrochemical performances; and the reasons for the difference are discussed. The experimental results indicate that the existence of NH4+ in the preparation solution has depressed the electrochemical performances of the final product; This is further confirmed by the electrochemical impedance spectra of the electrodes. 相似文献
The article describes a colorimetric assay for the determination of thrombin. It is based on the application of a triple enzyme-mimetic activity and a dual aptamer binding strategy. The triple signal amplification relies on oxidation of the chromogenic enzyme substrate 3,3,5,5-tetramethylbenzidine (TMB) that is catalyzed by composites consisting of graphene oxide (GO), gold/platinum nanoparticles (AuPtNP), and aptamer (Apt15), a G-quadruplex/hemin conjugate. The dual-aptamer target binding strategy is based on the fact that thrombin has two active sites to be recognized by its aptamers (Apt15 and Apt29). Magnetic beads (MBs) were modified with Apt29 (Apt29-MB) and then are bound by the GO-AuPtNP-Apt15/G-quadruplex/hemin composites. In the presence of thrombin, Apt29-MB and the GO-AuPtNP-Apt15/G-quadruplex/hemin composites form a sandwich-like superstructure. Thus, the absorbance increases due to the formation of TMB oxide produced by catalysis of the composites. Under optimized conditions, the absorbance at 450 nm increases linearly in the 0.30 to 100 nM thrombin concentration range, and the limit of detection is 0.15 nM. The method is simple, rapid, and does not require complicated instrumentation. Bovine serum albumin, human serum albumin and other proteins were found not to interfere.
Graphical abstract Schematic presentation of the photometric thrombin assay based on a triple enzyme-mimetic activity of combined nanomaterials (consisting of GO, AuPtNPs and the G-quadruplex/hemin DNAzyme) and two aptamers TMB: 3,3,5,5-tetramethylbenzidine, TMBox: 3,3,5,5-tetramethylbenzidine oxide, AuPtNP: gold/platinum nanoparticles).
We show here that the disubstituted ferrocenyl chalcones 1 and 2 are good electrochemical sensors for calcium and barium in CH3CN. However, these two triflate salts are detected in a different way by both ligands. To clarify this point, a thorough and informative NMR study of the ligand-salt interactions is presented. The unusual shapes of the titration curves obtained depend on both the ligand and cation used. For example, they illustrate that ligand 1 mainly interacts with the metal by its CO functions, while ligand 2 also interacts by its azacrown groups. These curves also reflect complex equilibriums in solution involving several ligand-salt adducts detected by mass spectrometry. To evaluate the strength of these interactions, the association constants of all the species formed have been determined by fitting the NMR data. It is noteworthy that changing the diethylamino groups in molecule 1 by the azacrown residue enhances the selectivity for the calcium salt, as pointed out by the value of the association constant of the 2Ca2+ species. The synthesis of the protonated counterparts 3 and 4 was useful to clarify the electrochemical behaviour of 1 and 2. Although the two ligand-salt interactions present several common points, the whole results obtained allow us to propose an original explanation for the difference observed between the Ca2+ and Ba2+ electrochemical sensing. 相似文献
Microemulsions have become a widely employed technique for the control of biodiesel quality but are still poorly understood as regard to their electrochemical behavior. In this work, we report the fundamental importance associated with the knowledge of electrochemical behavior of microemulsions composed of water in the presence of a supporting electrolyte, soybean biodiesel and propan‐1‐ol as consolute, along with the ferrocyanide–ferricyanide redox system applied as probe. The voltammetric results showed that for different compositions of microemulsions, variation in peak currents and change in system reversibility as well as in the electron transfer process were clearly noted. Furthermore, through the study of the diffusion coefficient, three different types of microemulsions including Oil/Water, Bicontinuous, and Water/Oil were successfully identified. Electrochemical impedance spectroscopy studies were also carried out aiming at obtaining more information regarding the electrode/solution interface. All the studies performed demonstrated that different types of microemulsions were formed upon exerting a direct influence on the electrochemical behavior of the redox probe. These results, in essence, point to the possibility of choosing a more suitable and advantageous microemulsion type for the development of an analytical method, as in the case, for example, of the microemulsions ME‐2 and ME‐3 which presented high voltammetric response in redox probe oxidation. 相似文献