RESUMO
Acute lymphoblastic leukemia (ALL) is one of the most prevalent cancers in children and microRNA-128 is amongst the most useful biomarkers not only for diagnosis of ALL, but also for discriminating ALL from acute myeloid leukemia (AML). In this study, a novel electrochemical nanobiosensor based on reduced graphene oxide (RGO) and gold nanoparticles (AuNPs) has been fabricated to detect miRNA-128. Cyclic Voltametery (CV), Square Wave Voltametery (SWV) and Electrochemical Impedance Spectroscopy (EIS) have been applied to characterize the nanobiosensor. Hexacyanoferrate as a label-free and methylene blue as a labeling material were used in the design of the nanobiosensors. It was found that the modified electrode has excellent selectivity and sensitivity to miR-128, with a limit of detection of 0.08761 fM in label-free and 0.00956 fM in labeling assay. Additionally, the examination of real serum samples of ALL and AML patients and control cases confirms that the designed nanobiosensor has the potential to detect and discriminate these two cancers and the control samples.
Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , MicroRNAs , Criança , Humanos , Ouro/química , Técnicas Eletroquímicas/métodos , Limite de Detecção , Nanopartículas Metálicas/química , Técnicas Biossensoriais/métodosRESUMO
Due to their high sensitivity, simplicity, portability, self-contained, and low cost, the development of electrochemical biosensors is a beneficial way to diagnose and anticipate many types of cancers. An electrochemical nanocomposite-based aptasensor is fabricated for the determination of miRNA-128 concentration as the acute lymphoblastic leukemia (ALL) biomarker for the first time. The aptamer chains were immobilized on the surface of the glassy carbon electrode (GCE) through gold nanoparticles/magnetite/reduced graphene oxide (AuNPs/Fe3O4/RGO). Fast Fourier transform infrared (FTIR), X-ray diffraction (XRD), vibrating sample magnetometer (VSM), and transmission electron microscopy (TEM) were used to characterize synthesized nanomaterials. Cyclic voltammetry (CV), square wave voltammetry (SWV), and electrochemical impedance spectroscopy (EIS) were used to characterize the modified GCE in both label-free and labeled methods. The results indicate that the modified working electrode has high selectivity and for miRNA-128 over other biomolecules. The hexacyanoferrate redox system typically operated at around 0.3 V (vs. Ag/AgCl), and the methylene blue redox system ran at about 0 V, were used as an electrochemical probe. The detection limit and linear detection range for hexacyanoferrate and methylene blue are 0.05346 fM, 0.1-0.9 fM, and 0.005483 fM, 0.01-0.09 fM, respectively. The stability and diffusion control analyses were performed as well. In both label-free and labeled methods, the modified electron showed high selectivity for miRNA-128. The use of methylene blue as a safer redox mediator caused miRNA-128 to be detected with greater accuracy at low potentials in PBS media. The findings also show the substantial improvement in detection limit and linearity by using reduced graphene oxide-magnetite-gold nanoparticles that can be verified by comparing with previous studies on the detection of other miRNAs.
RESUMO
In this work, graphite/gold nanoparticles (G/AuNPs) were synthesized through a facile chemical method, and its potential application for direct protein attachment for electrochemical detection of carbon monoxide (CO) was investigated. The preparation of G/AuNPs electrodes was optimized by synthesizing the nanoparticles in different concentration of HAuCl4.3H2O at various temperatures. The G/AuNPs electrode was subsequently modified by four types of mercaptopropionic acid, including 1-mercaptopropionic, 3-mercaptopropionic, 6-mercaptopropionic, and 11-mercaptopropionic acid, to achieve the best structure for protein attachment. Visible absorption and electrochemical studies showed that 3-mercaptopropionic acid possesses the best performance regarding the electrical conductivity between electrode and protein redox center. The cyclic voltammetry results revealed that the modified electrode has an appropriate performance for CO detection at very low concentrations while keeping a linear response. The limit of detection for the modified electrode was calculated to be about 0.2 ppb. Finally, the interactions of cytochrome C and carbon monoxides were simulated using molecular dynamics (MD), and the effect of protein conformation changes on the electrochemical signal was thoroughly examined. The simulation results suggested that the proposed electrochemical sensor has an acceptable performance for the detection of CO due to less fluctuation of amino acids near the protein chain in the presence of CO molecules.
Assuntos
Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Técnicas Eletroquímicas , Eletrodos , Ouro , Limite de DetecçãoRESUMO
Herein, an electrochemical label-free biosensor designed for the detection of glycated albumin (GA) using reduced graphene oxide/Au nanoparticles (RGO/AuNPs) modified by anti-GA aptamer. For fast and simple modification of the electrode, the aptamer chain was thiolated. Transition electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) techniques were used to the characterization of synthesized materials. Structural analysis of nanomaterials shows that graphene sheets were synthesized very fine by average thickness of 2.5 nm and Au nanoparticles distributed on the surface of graphene sheets uniformly. Cyclic voltammetry (CV) square wave voltammetry (SWV) and impedance spectroscopy (EIS) were used to electrochemical study of the decorated electrode. Electrochemical studies described the potential of fabricated rGO/AuNPs-aptamer electrode to selectively determine GA properly in buffer solution at the range of 2-10 µg mL-1 by the detection limit of 0.07 µg. mL-1 for GA.
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An electrochemical aptasensor is described for the voltammetric determination of lipopolysaccharide (LPS) from Escherichia coli 055:B5. Aptamer chains were immobilized on the surface of a glassy carbon electrode (GCE) via reduced graphene oxide and gold nanoparticles (RGO/AuNPs). Fast Fourier transform infrared, X-ray diffraction and transmission electron microscopy were used to characterize the nanomaterials. Cyclic voltammetry, square wave voltammetry and electrochemical impedance spectroscopy were used to characterize the modified GCE. The results show that the modified electrode has a good selectivity for LPS over other biomolecules. The hexacyanoferrate redox system, typically operated at around 0.3 V (vs. Ag/AgCl) is used as an electrochemical probe. The detection limit is 30 fg·mL-1. To decrease the electrochemical potential for detection of LPS, Mg/carbon quantum dots were used as redox active media. They decrease the detection potentialto 0 V and the detection of limit (LOD) to 1 fg·mL-1. The electrode was successfully used to analyze serum of patients and healthy persons. Graphical abstractSchematic representation of the modification of reduced graphene oxide gold nanoparticles with aptamer chains to immobilize on the glassy carbon electrode surface for electrochemical detection of lipopolysaccharides.
Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Escherichia coli/química , Lipopolissacarídeos/sangue , Nanopartículas Metálicas/química , Sequência de Bases , Eletrodos , Ferrocianetos/química , Ouro/química , Grafite/química , Humanos , Limite de Detecção , Magnésio/química , Oxirredução , Pontos Quânticos/químicaRESUMO
In this paper, a most sensitive electrochemical biosensor for detection of prostate-specific antigen (PSA) was designed. To reach the goal, a sandwich type electrode composed of reduced graphene oxide/ gold nanoparticles (GO/AuNPs), Anti-Total PSA monoclonal antibody, and anti-Free PSA antibody was assembled. The functionalized materials were thoroughly characterized by atomic force microscope spectroscopy, transmission electron microscopy, and X-ray diffraction techniques. The electrochemical properties of each of the modification step were evaluated by cyclic voltammetry and electrochemical impedance spectroscopy. The results presented that the proposed biosensor possesses high sensitivity toward total and free PSA. Furthermore, the fabricated biosensor revealed an excellent selectivity for PSA in comparison to the other tumor markers such as BHCG, Alb, CEA, CA125, and CA19-9. The limit of detection for the proposed electrochemical biosensor was estimated to be around 0.2 and 0.07 ng/mL for total and free PSA antigen, respectively.
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Cheese production is relied upon the action of Rennet on the casein micelles of milk. Chymosin assay methods are usually time consuming and offline. Herein, we report a new electrochemical technique for studying the proteolysis of K-casein. The interaction of rennet and its substrate was studied by fast Fourier transform continuous cyclic voltammetry (FFTCCV) based on a determination of k-casein in aqueous solution. FFTCCV technique is a very useful method for studying the enzymatic procedures. Fast response, no need of modified electrodes or complex equipment is some of FFTCCV advantages. Various concentrations of enzyme and substrate were selected and the increase in the appearance of charged species in solution as a result of the addition of rennet was studied. Data obtained using FFTCCV technique were also confirmed by turbidity analysis. The results show that rennet proteolysis activity occurs in much shorter time scales compare with its aggregation. Hence, following the appearance of charged segments as a result of proteolysis could be under consideration as a rapid and online method.
Assuntos
Caseínas/metabolismo , Análise de Fourier , Leite/química , Proteólise , Animais , Caseínas/química , EletroquímicaRESUMO
In this work, supercapacitive performance of polypyrrole copper chromate nano particles (Ppy/CuCr2O4 NPs) was studied. CuCr2O4 NPs with the average size of 20nm were synthesized simply by hydrothermal method and the composite electrodes were then electropolymerized on the surface of glassy carbon electrode. Common surface analysis techniques such as scanning electron microscopy (SEM), transmission electron microscopy(TEM) and Fourier transform infrared (FTIR) were used to study the morphology and structure of the composite. Furthermore, for electrochemical evaluation of composite electrodes, techniques including cyclic voltammetry (CV), galvanostatic charge discharge (CD) and impedance spectroscopy (EIS) were used. Using cyclic voltammetry, the specific capacitance values of Ppy and Ppy/CuCr2O4 NPs were calculated to be 109 and 508 F g-1, respectively. Results show that using CuCr2O4 NPs in the structure of polymeric films led to increased specific capacitance of composite electrodes more than four times that of poly pyrrole. Increasing the conductivity and stability of composite electrodes through continuous cycles are the other advantages of using CuCr2O4 NPs as active materials in a polymeric structure.