RESUMO
Atherosclerosis cardiovascular disease (ASCVD) has become one of the leading death causes in humans. Low-density lipoprotein (LDL) is an important biomarker for assessing ASCVD risk level. Thus, monitoring LDL levels can be an important means for early diagnosis of ASCVD. Herein, a novel electrochemical aptasensor for determination LDL was designed based on nitrogen-doped reduced graphene oxide-hemin-manganese oxide nanoparticles (NrGO-H-Mn3O4 NPs) integrated with clustered regularly interspaced short palindromic repeats and associated proteins (CRISPR/Cas12a) system. NrGO-H-Mn3O4 NPs not only have a large surface area and remarkable enhanced electrical conductivity but also the interconversion of different valence states of iron in hemin can provide an electrical signal. Nonspecific single-stranded DNA (ssDNA) was bound to NrGO-H-Mn3O4 NPs to form a signaling probe and was immobilized on the electrode surface. The CRISPR/Cas12a system has excellent trans-cleavage activity, which can be used to cleave ssDNA, thus detaching the NrGO-H-Mn3O4 NPs from the sensing interface and attenuating the electrical signal. Significant signal change triggered by the target was ultimately obtained, thus achieving sensitive detection of the LDL in range from 0.005 to 1000.0 nM with the detection limit of 0.005 nM. The proposed sensor exhibited good stability, selectivity, and stability and achieved reliable detection of LDL in serum samples, demonstrating its promising application prospects for the diagnostic application of LDL.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Sistemas CRISPR-Cas , Técnicas Eletroquímicas , Grafite , Hemina , Limite de Detecção , Lipoproteínas LDL , Compostos de Manganês , Óxidos , Compostos de Manganês/química , Lipoproteínas LDL/sangue , Lipoproteínas LDL/química , Humanos , Técnicas Eletroquímicas/métodos , Óxidos/química , Grafite/química , Aptâmeros de Nucleotídeos/química , Hemina/química , Técnicas Biossensoriais/métodos , DNA de Cadeia Simples/química , Nanopartículas/químicaRESUMO
Golgi protein 73 (GP73) is a novel tumor marker in the early diagnosis and prognosis of hepatocellular carcinoma (HCC). Herein, a competitive electrochemical aptasensor for detecting GP73 was constructed using reduced graphene oxide-ferrocene-polyaniline nanocomposite (rGO-Fc-PANi) as the biosensing platform. The rGO-Fc-PANi had larger specific surface area, excellent conductivity and outstanding electroactive performance, which served as nanocarrier for GP73 aptamer (GP73Apt) binding and as redox nanoprobe for record electrical signal. Then, a complementary chain (cDNA) was fixed to the electrode by hybridization with GP73Apt. When GP73 was present, a competitive process happened among cDNA, GP73Apt and GP73, formed the GP73-GP73Apt stable chemical structure and made cDNA detach from the sensing electrode, resulting in enhancement of electrical signal. The difference in the corresponding peak current before and after the competition can be used to indicate the quantitative of GP73. Under optimal conditions, the DPV current response showed a good log-linear relationship with GP73 concentrations (0.001 â¼ 100.0 ng/mL) with a detection limit of 0.15 pg/mL (S/N = 3). It was successfully used for GP73 detection in human serum with RSDs ranging from 1.08 % to 6.96 %. Therefore, the aptasensor could provide an innovative technology platform and hold a great potential in clinical application.
RESUMO
A new sandwich-type electrochemical biosensing platform was developed by gold @polyphthalenediamine nanohybrids (AuNP@PoPD) as the sensing platform and phosphorus doped reduced graphene oxide-hemin-palladium nanoparticles (PrGO-Hemin-PdNP) as the signal amplifier for phosphatidylinositol proteoglycan 3 (GPC3). AuNP@PoPD, co-electrodeposited into the screen printed electrode with high conductivity and stability, is dedicated to assembling the primary GPC3 aptamer (GPC3Apt). The second GPC3Apt immobilized on the high conductivity and large surface area of PrGO-Hemin-PdNP was utilized as an electrochemical signal reporter by hemin oxidation (PrGO-Hemin-PdNP-GPC3Apt). In the range 0.001-10.0 ng/mL, the hemin oxidation current signal of the electrochemical aptasensor increased log-linearly with the concentration of GPC3, the lowest detection limit was 0.13 pg/mL, and the sensitivity was 2.073 µA/µM/cm2. The aptasensor exhibited good sensing performance in a human serum sample with the relative error of 4.31-8.07%. The sandwich sensor showed good selectivity and stability for detection GPC3 in human serum samples, providing a new efficient and sensitive method for detecting HCC markers.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Técnicas Eletroquímicas , Glipicanas , Ouro , Grafite , Hemina , Limite de Detecção , Nanopartículas Metálicas , Paládio , Glipicanas/sangue , Humanos , Técnicas Eletroquímicas/métodos , Técnicas Eletroquímicas/instrumentação , Aptâmeros de Nucleotídeos/química , Hemina/química , Grafite/química , Paládio/química , Ouro/química , Técnicas Biossensoriais/métodos , Nanopartículas Metálicas/química , EletrodosRESUMO
Glypican-3 (GPC3) is an essential reference target for hepatocellular carcinoma detection, follow-up and prediction. Herein, a dual-signal electrochemical aptasensor based on reduced graphene oxide-cuprous oxide (RGO-Cu2O) nanozyme was developed for GPC3 detection. The RGO-Cu2O nanoenzyme displayed excellent electron transport effect, large specific surface area and outstanding peroxidase-like ability. The differential pulse voltammetry (DPV) signal of Cu2O oxidation fraction and the chronoamperometry (i-t) signal of H2O2 decomposition catalyzed by RGO-Cu2O nanozyme were used as dual-signal detection. Under optimal conditions, the log-linear response ranges were 0.1 to 500.0 ng/mL with the limit of detection 0.064 ng/mL for DPV technique, and 0.1-50.0 ng/mL for i-t technique (detection limit of 0.0177 ng/mL). The electrochemical aptasensor has remarkably analytical performance with wide response range, low detection limit, excellent repeatability and specificity, good recovery in human serum samples. The two output signals of one sample achieve self-calibration of the results, effectively avoiding the occurrence of possible leakage and misdiagnosis of a single detection signal, suggesting that it will be a promising method in the early biomarker detection.
Assuntos
Técnicas Biossensoriais , Cobre , Técnicas Eletroquímicas , Glipicanas , Grafite , Limite de Detecção , Grafite/química , Glipicanas/sangue , Glipicanas/análise , Humanos , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Cobre/química , Aptâmeros de Nucleotídeos/química , Catálise , Oxirredução , Peróxido de Hidrogênio/químicaRESUMO
Golgi protein 73 (GP73) is a new serum marker associated with early diagnosis and postoperative assessment of hepatocellular carcinoma (HCC). Herein, an electrochemical/fluorescence dual-signal biosensor was designed for determination of GP73 based on molybdenum disulfide/ferrocene/palladium nanoparticles (MoS2-Fc-PdNPs) and nitrogen-doped graphene quantum dots (NGQDs). GP73 aptamer (Apt) was labeled with NGQDs to form the NGQDs-Apt fluorescence probe. MoS2-Fc-PdNPs served not only as the fluorescence quencher but also as electrochemical enhancer. The sensing platform (NGQDs-Apt/MoS2-Fc-PdNPs) was formed based on the fluorescence resonance energy transfer (FRET) mechanism. In the presence of GP73, the specific binding of NGQDs-Apt to GP73 interrupted FRET, restoring the fluorescence of NGQDs-Apt at λex/em = 348/438 nm and enhancing the oxidation current of Fc in MoS2-Fc-PdNPs at 0.04 V through differential pulse voltammetry (DPV). Under the optimal conditions, the DPV current change and fluorescence recovery have a good linear relationship with GP73 concentration from 1.00 to 10.0 ng/mL. The calibration equation for the fluorescence mode was Y1 = (0.0213 ± 0.00127)X + (0.0641 ± 0.00448) and LOD was 0.812 ng/mL (S/N = 3). The calibration equation of the electrochemical mode was Y2 = (3.41 ± 0.111)X + (1.62 ± 0.731), and LOD of 0.0425 ng/mL (S/N = 3). The RSDs of fluorescence mode and electrochemical mode after serum detection were 1.62 to 5.21% and 0.180 to 6.62%, respectively. By combining the electrochemical and fluorescence assay, more comprehensive and valuable information for GP73 was provided. Such dual-mode detection platform shows excellent reproducibility, stability, and selectivity and has great application potential.
Assuntos
Carcinoma Hepatocelular , Dissulfetos , Grafite , Neoplasias Hepáticas , Nanopartículas Metálicas , Pontos Quânticos , Humanos , Molibdênio , Paládio , Nitrogênio , Reprodutibilidade dos Testes , MetalocenosRESUMO
1,5-anhydroglucitol (1,5-AG) is of considerable clinical relevance as a biochemical marker of glucose metabolism in the assessment and monitoring of diabetes. Herein, a simple colorimetric biosensor was constructed for the identification and detection of 1,5-AG by using pyranose oxidase (PROD) enzyme cascaded with reduced graphene oxide/persimmon tannin/Pt@Pd (RGO-PT/Pt@Pd NPs) nanozyme. The as-prepared RGO-PT/Pt@Pd NPs had excellent peroxidase-like activity and can be applied as a nanozyme. First, PROD enzyme reacts with the target 1,5-AG, decomposing 1,5-AG into 1,5-anhydrofuctose (1,5-AF) and H2O2. At this point, the highly catalytic RGO-PT/Pt@Pd NPs nanozyme produces a cascade with PROD enzyme which catalyzes the decomposition of H2O2 to produce O2. This in turn oxidizes the substrate 3,3',5,5'-tetramethylbenzidine (TMB) and produces a color change in the solution. Finally, the detection of 1,5-AG was achieved by measuring the absorption peak at 652 nm with an ultraviolet visible (UV-vis) spectrophotometer. Under optimal conditions, the linear operating range of the 1,5-AG enzyme cascade colorimetric sensor was 1.0-100.0 µg/mL, and the limit of detection (LOD) was 0.81 µg/mL. The proposed colorimetric biosensor was successfully applied to detect 1,5-AG in spiked human serum samples with the recoveries of 97.2-103.9% and RSDs of 1.94-4.48%. It provides a promising developmental assay for clinical detection of 1,5-AG.
Assuntos
Diospyros , Peróxido de Hidrogênio , Humanos , Peróxido de Hidrogênio/química , Diospyros/metabolismo , Colorimetria , Taninos , Citocromo P-450 CYP2B1 , Peroxidase/químicaRESUMO
Breeze energy is a widely distributed renewable energy source in the natural world, but its efficient exploitation is very difficult. The conventional harvester with fixed arm length (HFA) has a relatively high start-up wind speed owing to its high and constant rotational inertia. Therefore, this paper proposes a harvester with a helix s-type vertical axis (HSVA) for achieving random energy capture in the natural breeze environment. The HSVA is constructed with two semi-circular buckets driven by the difference of the drag exerted, and the wind energy is transferred into mechanical energy. Firstly, as the wind speed changes, the HSVA harvester can match the random breeze to obtain highly efficient power. Compared with the HFA harvester, the power coefficient is significantly improved from 0.15 to 0.2 without additional equipment. Furthermore, it has more time for energy attenuation as the wind speeds dropped from strong to moderate. Moreover, the starting torque is also better than that of HFA harvester. Experiments showed that the HSVA harvester can improve power performance on the grounds of the wind speed ranging in 0.8-10.1 m/s, and that the star-up wind speed is 0.8 m/s and output peak power can reach 17.1 mW. In comparison with the HFA harvester, the HSVA harvester can obtain higher efficient power, requires lower startup speed and keeps energy longer under the same time. Additionally, as a distributed energy source, the HSVA harvester can provide a self-generating power supply to electronic sensors for monitoring the surrounding environment.
RESUMO
1,5-Anhydroglucitol (1,5-AG) is a sensitive biomarker for real-time detection of diabetes mellitus. In this study, an electrochemical biosensor to specifically detect 1,5-AG levels based on persimmon-tannin-reduced graphene oxide-PtPd nanocomposites (PT-rGO-PtPd NCs), which were modified onto the surface of a screen-printed carbon electrode (SPCE), was designed. The PT-rGO-PtPd NCs were prepared by using PT as the film-forming material and ascorbic acid as the reducing agent. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), ultraviolet-visible spectroscopy (UV-vis), and X-ray diffraction (XRD) spectroscopy analysis were used to characterise the newly synthesised materials. PT-rGO-PtPd NCs present a synergistic effect not only to increase the active surface area to bio-capture more targets, but also to exhibit electrocatalytic efficiency to catalyze the decomposition of hydrogen peroxide (H2O2). A sensitive layer is formed by pyranose oxidase (PROD) attached to the surface of PT-rGO-PtPd NC/SPCE. In the presence of 1,5-AG, PROD catalyzes the oxidization of 1,5-AG to generate 1,5-anhydrofuctose (1,5-AF) and H2O2 which can be decomposed into H2O under the synergistic catalysis of PT-rGO-PtPd NCs. The redox reaction between PT and its oxidative product (quinones, PTox) can be enhanced simultaneously by PT-rGO-PtPd NCs, and the current signal was recorded by the differential pulse voltammetry (DPV) method. Under optimal conditions, our biosensor shows a wide range (0.1-2.0 mg/mL) for 1,5-AG detection with a detection limit of 30 µg/mL (S/N = 3). Moreover, our electrochemical biosensor exhibits acceptable applicability with recoveries from 99.80 to 106.80%. In summary, our study provides an electrochemical method for the determination of 1,5-AG with simple procedures, lower costs, good reproducibility, and acceptable stability.
RESUMO
Glypican-3 (GPC3), as an emerging biomarker, has been shown to be beneficial for the early diagnosis and treatment of hepatocellular carcinoma (HCC). In this study, an ultrasensitive electrochemical biosensor for GPC3 detection has been constructed based on the hemin-reduced graphene oxide-palladium nanoparticles (H-rGO-Pd NPs) nanozyme-enhanced silver deposition signal amplification strategy. When GPC3 specifically interacted with GPC3 antibody (GPC3Ab) and GPC3 aptamer (GPC3Apt), an "H-rGO-Pd NPs-GPC3Apt/GPC3/GPC3Ab" sandwich complex was formed with peroxidase-like properties which enhanced H2O2 to reduce the silver (Ag) ions in solution to metallic Ag, resulting in the deposition of silver nanoparticles (Ag NPs) on the surface of the biosensor. The amount of deposited Ag, which was derived from the amount of GPC3, was quantified by the differential pulse voltammetry (DPV) method. Under ideal circumstances, the response value was linearly correlated with GPC3 concentration at 10.0-100.0 µg/mL with R2 of 0.9715. When the GPC3 concentration was in the range from 0.01 to 10.0 µg/mL, the response value was logarithmically linear with the GPC3 concentration with R2 of 0.9941. The limit of detection was 3.30 ng/mL at a signal-to-noise ratio of three and the sensitivity was 1.535 µAµM-1cm-2. Furthermore, the electrochemical biosensor detected the GPC3 level in actual serum samples with good recoveries (103.78-106.52%) and satisfactory relative standard deviations (RSDs) (1.89-8.81%), which confirmed the applicability of the sensor in practical applications. This study provides a new analytical method for measuring the level of GPC3 in the early diagnosis of HCC.
Assuntos
Técnicas Biossensoriais , Glipicanas , Grafite , Nanopartículas Metálicas , Humanos , Técnicas Biossensoriais/métodos , Carcinoma Hepatocelular , Técnicas Eletroquímicas/métodos , Grafite/química , Hemina/química , Peróxido de Hidrogênio , Neoplasias Hepáticas , Nanopartículas Metálicas/química , Paládio , Prata/químicaRESUMO
The effective detection of biomarkers associated with hepatocellular carcinoma (HCC) is of great importance. Golgi protein 73 (GP73), a serum biomarker of HCC, has better diagnostic value than Alpha-fetoprotein (AFP) has been reported. In this paper, highly accurate fluorescence sensing platform for detecting GP73 was constructed based on fluorescence resonance energy transfer (FRET), in which nitrogen-doped graphene quantum dots (NGQDs) labelling with GP73 aptamer (GP73Apt) was used as fluorescence probe, and molybdenum disulfide @ reduced graphene oxide (MoS2@RGO) nanosheets was used as fluorescent receptors. MoS2@RGO nanosheets can quench the fluorescence of NGQDs-GP73Apt owing to FRET mechanisms. In the presence of GP73, the NGQDs-GP73Apt specifically bound with GP73 to from the deployable structures, making NGQDs-GP73Apt far away from MoS2@RGO nanosheets, blocking the FRET process, resulting in fluorescence recovery of NGQDs-GP73Apt. Under optimal conditions, the recovery intensity of fluorescence in the detection system is linearly related to the concentration of GP73 in the range of 5 ng/mL - 100 ng/mL and the limit of detection is 4.54 ng/mL (S/N = 3). Moreover, detection of GP73 was performed in human serum samples with good recovery (97.21-100.83%). This platform provides a feasible method for the early diagnosis of HCC, and can be easily extended to the detection of other biomarkers.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Carcinoma Hepatocelular , Grafite , Neoplasias Hepáticas , Pontos Quânticos , Humanos , Pontos Quânticos/química , Grafite/química , Molibdênio/química , Nitrogênio/química , Neoplasias Hepáticas/diagnóstico , Óxidos de Nitrogênio , Aptâmeros de Nucleotídeos/química , Óxido Nítrico , Técnicas Biossensoriais/métodosRESUMO
Glypican-3 (GPC3) is a membrane-associated proteoglycan that is specifically upregulated in hepatocellular carcinoma (HCC) and has become one of the most promising biomarkers closely related to the occurrence and development of HCC. In this work, platinum@palladium nanoparticles decorated with hemin-reduced graphene oxide (H-rGO-Pt@Pd NPs) were used not only as a support for GPC3 aptamer (GPC3Apt) immobilization, but also as a new redox nanoprobe in electrochemical analysis for the determination of GPC3. The electrochemical aptasensor involved a reaction cell with a three-electrode system, and the differential pulse voltammetry (DPV) technique was adopted. In the presence of GPC3, the formed GPC3Apt-GPC3 complexes had stable structures and were cleaved from the electrode surface, leading to more electroactive H-rGO-Pt@Pd NPs repelling freely from the GPC3Apt/H-rGO-Pt@Pd NPs and thus to the increase of the oxidation peak current of hemin in H-rGO-Pt@Pd NPs. Under optimal conditions and a working voltage of +700 mV (vs. Ag/AgCl), the label-free electrochemical GPC3 aptasensor showed superior performance with a wider concentration linear range (0.001-10.0 µg mL-1), a lower limit of detection (LOD) (0.181 ng mL-1, S/N = 3), a higher sensitivity (0.0446 µA µM-1 cm-2) and good selectivity. Furthermore, the fabricated aptasensor was applied to GPC3 determination in human serum samples with satisfactory recoveries of 94.3%-119% and RSDs of 0.15%-5.78%. The current work provides a flexible approach for the rapid and sensitive analysis of GPC3 and has a broad application prospect in the diagnosis of HCC.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Carcinoma Hepatocelular , Grafite , Neoplasias Hepáticas , Nanopartículas Metálicas , Humanos , Platina/química , Paládio/química , Hemina , Nanopartículas Metálicas/química , Técnicas Eletroquímicas/métodos , Glipicanas , Grafite/química , Técnicas Biossensoriais/métodos , Aptâmeros de Nucleotídeos/químicaRESUMO
A Golgi protein 73 (GP73) colorimetric biosensor based on the reduced graphene oxide-carboxymethyl chitosan-hemin/platinum@palladium nanoparticles (RGO-CMCS-Hemin/Pt@Pd NPs) with peroxidase-like activity was constructed. The RGO-CMCS-Hemin/Pt@Pd NPs with high peroxidase-like activity were successfully synthesized under mild conditions. Then, the aminylated GP73 aptamer (Apt) was bound to the RGO-CMCS-Hemin/Pt@Pd NPs to form the recognition probe. Another unmodified GP73 aptamer (AptI) was served as the capture probe. In the presence of target GP73, the capture probe and the recognition probe specifically bind to GP73 and form a RGO-CMCS-Hemin/Pt@Pd NP-Apt/GP73/AptI sandwich-type structure, which can oxidase the colorless 3,3',5,5'-tetramethylbenzidine (TMB) into blue oxTMB in the presence of H2O2. GP73 detection was achieved by measuring the peak UV absorption at 652 nm. Under the optimum conditions, the GP73 concentration was linearly related to the absorbance intensity in the range 10.0-110.0 ng/mL, and the limit of detection (LOD) was 4.7 ng/mL. The proposed colorimetric biosensor was successfully applied to detect GP73 in spiked human serum samples with recoveries of 98.2-107.0% and RSDs of 1.90-5.44%, demonstrating the excellent potential for highly sensitive GP73 detection in clinical detection. A colorimetric biosensor for visual determination of GP73 based on RGO-CMCS-Hemin/Pt@Pd NPs nanozyme with peroxidase-like activity was designed. The GP73 biosensor responses linearly from 10.0-110.0 ng/mL with LOD of 4.7 ng/mL, and shows acceptable specificity and good recovery.
Assuntos
Técnicas Biossensoriais , Quitosana , Nanopartículas Metálicas , Quitosana/química , Colorimetria , Dimaprit/análogos & derivados , Grafite , Hemina , Humanos , Peróxido de Hidrogênio/química , Nanopartículas Metálicas/química , Paládio/química , Peroxidase/química , Peroxidases , Platina/químicaRESUMO
Golgi protein 73 (GP73) is a new type of marker that can specifically detect hepatocellular carcinoma (HCC). Herein, a dual-signal sandwich-type electrochemical aptasensor for GP73 determination was constructed on the basis of hemin-reduced graphene oxide-manganese oxide (H-rGO-Mn3O4) nanozymes. Gold@poly(o-phenylenediamine) (Au@POPD) nanohybrids with a large specific surface area and conductance were co-electro-deposited onto a screen-printed electrode (SPE) surface to immobilize GP73 capture aptamer 2 (Apt2). H-rGO-Mn3O4 nanozymes were used not only to immobilize amino functionalised GP73 aptamer 1 (Apt1) as the detection probe, but also to serve as an in-situ redox signal indicator because of the redox reaction of Hemin (Fe(Ш)/Hemin(Fe(II)). In addition, given their excellent peroxidase-like activity, H-rGO-Mn3O4 nanozymes can catalyse the decomposition of H2O2 and oxidation of substrate (3,3',5,5'-tetramethylbenzidine, TMB) to oxTMB, which is used as another redox signal. In the presence of the target GP73, the two aptamers specifically bind to the target, thereby affecting two electrochemical signals. Under optimal conditions, the dual-signal sandwich-type electrochemical aptasensor had a salient analytical performance. The two electrochemical redox signals linearly increase with the logarithm of the GP73 concentration in the range of 0.01-100.0 ng/mL with the limit of detection (LOD) of 0.0071 ng/mL and sensitivity of 2.441 µA/µM/cm2. Moreover, the recovery of human serum samples ranged from 98.66% to 121.11%. Furthermore, the two redox signals can simultaneously corroborate each other, thereby preventing missed diagnosis and misdiagnosis. All the results can provide new insights into the clinically effective determination of HCC.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Carcinoma Hepatocelular , Grafite , Neoplasias Hepáticas , Nanopartículas Metálicas , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Carcinoma Hepatocelular/diagnóstico , Técnicas Eletroquímicas/métodos , Ouro/química , Grafite/química , Hemina/química , Humanos , Peróxido de Hidrogênio/química , Limite de Detecção , Neoplasias Hepáticas/diagnóstico , Nanopartículas Metálicas/químicaRESUMO
Diabetes is one of metabolic diseases affecting major human health. The early diagnosis and treatment of diabetes have significant benefits. 1,5-anhydroglucitol (1,5-AG) accurately reflects a patient's average blood glucose level for the past 3-7 days and becomes a promising marker for real-time detection of diabetes. In this study, a novel biosensor for determination 1,5-AG is constructed using reduce graphene oxide-carboxymethylated chitosan-hemin@platinum nanocomposites (rGO-CMC-H@Pt NCs) nanozyme and pyranose oxidase (PROD) enzyme as the electrochemical biosensing platform. The rGO-CMC-H@Pt NCs nanozyme has good electro-conductibility, high specific surface area, and admirable peroxide-like catalysis effect to enhance the electrochemical response. 1,5-AG is catalyzed by PROD and produces hydrogen peroxide (H2O2), which in turn can be decomposed by rGO-CMC-H@Pt NCs and produce a current signal recorded by differential pulse voltammetry (DPV) technique. Under optimal conditions, the response currents have a linear relationship in the 1,5-AG concentration of 0.1-2.0 mg/mL with R2 of 0.9869. The sensitivity is 2.1895 µA/µg·mL-1 and the limit of detection (LOD) is 38.2 µg/mL (S/N = 3). In addition, the specificity, reproducibility, stability and recovery (94.5-107.6%) of 1,5-AG biosensors all exhibit good performance. Therefore, the designed 1,5-AG biosensor has a good effect and can be used for the diagnosis of diabetes.
Assuntos
Técnicas Biossensoriais , Quitosana , Grafite , Técnicas Biossensoriais/métodos , Citocromo P-450 CYP2B1 , Desoxiglucose , Técnicas Eletroquímicas/métodos , Hemina , Humanos , Peróxido de Hidrogênio , Limite de Detecção , Platina , Reprodutibilidade dos TestesRESUMO
Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related deaths in China. Glypican-3 (GPC3) is a specific antigen related to HCC, which is widely used in clinical detection as a reliable marker of HCC. In this paper, a highly sensitive homogeneous apatasensor was designed for GPC3 detection based on fluorescence resonance energy transfer (FRET) where the GPC3 aptamer labelled gold carbon dots (AuCDs-GPC3Apt) are used as a donor and magnetic graphene oxide (Fe3O4/GO) nanosheets are used as an acceptor. A one-step hydrothermal method was used to synthesize AuCDs to provide sufficient fluorescence. The FRET phenomenon exists between AuCDs-GPC3Apt and Fe3O4/GO, which weakens the fluorescence intensity of the whole system. When the target GPC3 is added to the FRET system, the fluorescent AuCDs-GPC3Apt binds to the GPC3 and forms a folded structure, which leads to AuCDs-GPC3Apt separation from Fe3O4/GO nanosheets. The Fe3O4/GO is then magnetically separated so that the fluorescence of free labelled AuCDs-GPC3Apt is restored. Under the optimum conditions, the fluorescence recovery rate is linearly correlated with the concentration of GPC3 (5-100 ng·mL-1) and the detection limit is 3.01 ng·mL-1 (S/N = 3). This strategy shows recoveries from 98.76 to 101.29% in real human serum samples and provides an immediate and effective detection method for the quantification of GPC3 with great potential applications for early diagnosis of HCC. A sensitive homogeneous FRET-based apatasensor was designed for GPC3 detection where the AuCDs-GPC3Apt is a donor and Fe3O4/GO nanosheets are an acceptor. The GPC3 fluorescent aptasensor combines wider output range with low cost, high specificity, and good anti-interference.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Carcinoma Hepatocelular , Grafite , Neoplasias Hepáticas , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Carbono/química , Carcinoma Hepatocelular/diagnóstico , Detecção Precoce de Câncer , Transferência Ressonante de Energia de Fluorescência/métodos , Glipicanas , Ouro/química , Grafite/química , Humanos , Limite de Detecção , Neoplasias Hepáticas/diagnósticoRESUMO
In this paper, a novel silicon-based light-addressable potentiometric sensor (LAPS) has been designed for the detection of 1,5-anhydroglucitol (1,5-AG) in human serum. Reduced graphene oxide-chitosan-ferrocene (RGO-CS-Fc)/AuNPs nanohybrids and pyranose oxidase (PROD) enzyme is used to fabricate biological sensitive membrane unit by layer-by-layer assembly technology. When a bias voltage is provided to the LAPS system, the catalytic oxidation reaction between 1,5-AG and PROD to produce H2O2. The by-product H2O2 can oxidize Fc(Fe2+) ions in RGO-CS-Fc nanohybrids into Fc(Fe3+) ions, which cause the potential of the sensitive membrane surface to change and the potential shift of I-V curve will generate a corresponding offset response. Under the optimal conditions, the potential shift of the LAPS is linearly related to the concentration of 1,5-AG at 10⯵g·mL-1 -350⯵g·mL-1 with the correlation coefficient of 0.97414. The sensitivity is 0.44273â¯mV/µg·mL-1 and the lowest detection limit is 10⯵g·mL-1. In addition, the biosensor showed good specificity, acceptable stability and satisfactory recovery rates (91.28%-107.66%), which would be a potential testing methods in actual clinical samples.
Assuntos
Técnicas Biossensoriais/métodos , Desoxiglucose/sangue , Técnicas Eletroquímicas/métodos , Nanopartículas Metálicas/química , Ouro/química , HumanosRESUMO
An electrochemical aptasensor for quantitatively detecting glypican-3 (GPC3) was constructed by combining hemin-reduced graphene oxide-platinum (H-rGO-Pt) nanoparticles (NPs) with reduced graphene oxide-gold (rGO-Au) nanoparticles (NPs). Herein, the rGO-Au NPs deposited onto screen-printed electrodes resulted in signal amplification due to their large surface areas. Meanwhile, highly conductive H-rGO-Pt NPs acted as a sensing medium that improved electrical conductivity and as an indicator for monitoring peak current for determination. A GPC3 aptamer (GPC3apt) with a low equilibrium dissociation constant was used as a bio-recognition molecule. GPC3apt specifically captured GPC3 proteins and formed aptamer-GPC3 complexes, which impeded electron transfer and thus hampered the redox signal of hemin in H-rGO-Pt NPs. This developed electrochemical aptasensor showed a linear response to GPC3 (from 0.001 µg/mL to 10 µg/mL) and had a detection limit of 0.001 µg/mL. This work provides a low-cost and highly sensitive detection with and good recovery for GPC3 and holds great promise for the clinical diagnosis of hepatocellular carcinoma.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Condutividade Elétrica , Técnicas Eletroquímicas , Glipicanas , Ouro , Hemina , Limite de Detecção , PlatinaRESUMO
Glypican-3 (GPC3) is a highly specific tumor marker for hepatocellular carcinoma (HCC), and plays an important role in reflecting the existence, therapeutic evaluation, monitoring and prognosis of HCC. Herein, an electrochemical aptasensor was designed for GPC3 detection with the reduced graphene oxide-hemin nanocomposites (RGO-Hemin) modified on the screen-printed electrode surface as the sensing platform and GPC3 aptamer as recognize molecule. In the existence of GPC3, the aptamer can specifically bind with the target GPC3 and form GPC3-aptamer conjugations on the sensing surface, which would increase the resistance of the electron transfer on the electrode and make the decrease of electrochemical signals of Hemin in RGO-Hemin nanocomposites. The electrochemical current change was recorded by differential pulse voltammetry (DPV). Scanning electron microscopy (SEM), Raman microscope (RM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to characterize the GPC3 electrochemical aptasensor. Under the optimum conditions, the current response of the electrochemical aptasensor is linearly correlated with the concentration of GPC3 (0.001-10.0 µg/mL) with the detection limit of 2.86 ng/mL (S/N = 3) and the sensitivity of 0.134 µA/µM/cm2. In addition, the aptasensor was applied to the determination of GPC3 in spiked human plasma and the recoveries fluctuated from 102.68% to 117.29%. All these results show that the aptasensor has good specificity, sensitivity, stability and reproducibility for GPC3 detection.
Assuntos
Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Glipicanas/análise , Grafite/química , Hemina/química , Limite de Detecção , Nanocompostos/química , Eletroquímica , Eletrodos , Glipicanas/sangue , Glipicanas/química , Humanos , Oxirredução , Impressão , Propriedades de SuperfícieRESUMO
Herein, a label-free electrochemical aptasensor for alpha-fetoprotein (AFP) analysis was established. The AFP aptamer (AFP-Apt), as the recognition molecule, was immobilized on the surface of a screen-printed carbon electrode, which was modified by gold-platinum metallic nanoparticles and reduced graphene oxide-chitosan-ferrocene nanohybrids (Au-Pt NPs/RGO-CS-Fc), to build the AFP electrochemical aptasensor. The construction process of the aptasensor was characterized by scanning electron microscopy, X-ray photoelectron spectroscopy, Raman spectroscopy, and electrochemical impedance spectroscopy. With the addition of AFP, the formation of the AFP-aptamer conjugation blocked the electron transfer reaction, reducing the differential pulse voltammetric responses of the current of Fc in the RGO-CS-Fc nanohybrids. By optimizing the experimental parameters, AFP could be detected with the dynamic concentration range of 0.001 to 10.0 µg mL-1 and with a detection limit of 0.3013 ng mL-1. In addition, the approach was manifested to have good selectivity, reproducibility, and stability. The fabricated aptasensor had a good recovery rate of 102.36% to 118.09% in real human serum samples. This work demonstrates that the electrochemical aptasensor is a useful tool for analyzing AFP inexpensively, rapidly, and accurately.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Nanopartículas Metálicas , Nanocompostos , Técnicas Eletroquímicas , Ouro , Grafite , Humanos , Platina , Reprodutibilidade dos Testes , alfa-FetoproteínasRESUMO
A Glypican-3 (GPC3) electrochemical aptamer nanobiosensor based on hemin/graphene nanohybrids (HGNs) peroxidase-like catalytic silver deposition and GPC3 aptamer has been constructed for the determination of GPC3. The HGNs were prepared by an one-step reduction method. Fourier transform infrared spectroscopy (FT-IR), ultraviolet spectroscopy (UV-vis), and transmission electron microscopy (TEM) were used to study the structure and morphological characteristics of the HGNs. The GPC3 electrochemical aptamer nanobiosensor was constructed using HGNs-aptamer (HGNs-Apt) as the signal probe and GPC3 aptamer as the capture probe. With the help of the catalytic action of peroxidase-like properties of HGNs, H2O2 reduces the silver (Ag) ions in solution to metallic Ag, which deposit on the surface of the electrode. The amount of deposited Ag, which was derived from the amount of GPC3, was quantified by differential pulse voltammetry (DPV). Under optimal conditions, the current response of Ag had a good positive correlation with the GPC3 concentration in the range 10.0-100.0 µg mL-1 with a correlation coefficient of 0.9958. The detection limit was 3.16 µg mL-1 at a signal-to-noise ratio of 3, and the sensitivity was calculated to be 0.807 µA µM-1 cm-2. The method is validated by analyzing spiked human serum samples with good recovery ranging from 101 to 122%. In addition, the GPC3 electrochemical aptamer nanobiosensor has acceptable selectivity, stability, and reproducibility. Graphical abstract A Glypican-3 electrochemical aptamer nanobiosensor based on hemin/graphene nanohybrids (HGNs) peroxidase-like catalytic silver deposition and GPC3 aptamer has been constructed for the determination of GPC3. The electrochemical aptamer nanobiosensor exhibits high selectivity, acceptance reproducibility, and good recovery performances.