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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.
Asunto(s)
Aptámeros de Nucleótidos , Técnicas Biosensibles , Sistemas CRISPR-Cas , Técnicas Electroquímicas , Grafito , Hemina , Límite de Detección , Lipoproteínas LDL , Compuestos de Manganeso , Óxidos , Compuestos de Manganeso/química , Lipoproteínas LDL/sangre , Lipoproteínas LDL/química , Humanos , Técnicas Electroquímicas/métodos , Óxidos/química , Grafito/química , Aptámeros de Nucleótidos/química , Hemina/química , Técnicas Biosensibles/métodos , ADN de Cadena Simple/química , Nanopartículas/químicaRESUMEN
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.
RESUMEN
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.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Técnicas Electroquímicas , Glipicanos , Oro , Grafito , Hemina , Límite de Detección , Nanopartículas del Metal , Paladio , Glipicanos/sangre , Humanos , Técnicas Electroquímicas/métodos , Técnicas Electroquímicas/instrumentación , Aptámeros de Nucleótidos/química , Hemina/química , Grafito/química , Paladio/química , Oro/química , Técnicas Biosensibles/métodos , Nanopartículas del Metal/química , ElectrodosRESUMEN
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.
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Técnicas Biosensibles , Cobre , Técnicas Electroquímicas , Glipicanos , Grafito , Límite de Detección , Grafito/química , Glipicanos/sangre , Glipicanos/análisis , Humanos , Técnicas Electroquímicas/métodos , Técnicas Biosensibles/métodos , Cobre/química , Aptámeros de Nucleótidos/química , Catálisis , Oxidación-Reducción , Peróxido de Hidrógeno/químicaRESUMEN
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.
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Carcinoma Hepatocelular , Disulfuros , Grafito , Neoplasias Hepáticas , Nanopartículas del Metal , Puntos Cuánticos , Humanos , Molibdeno , Paladio , Nitrógeno , Reproducibilidad de los Resultados , MetalocenosRESUMEN
The detection of serum markers is important for the early diagnosis and monitoring of diseases, but conventional detection methods have the problem of low specificity or sensitivity. CRISPR/Cas13a-based biosensors have the characteristics of simple detection methods and high sensitivity, which have a certain potential to solve the problems of conventional detection. This paper focuses on the research progress of CRISPR/Cas13a-based biosensors in serum marker detection, introduces the principles and applications of fluorescence, electrochemistry, colorimetric, and other biosensors based on CRISPR/Cas13a in the detection of serum markers, compares and analyzes the differences between the above CRISPR/Cas13a-based biosensors, and looks forward to the future development direction of CRISPR/Cas13a-based biosensors.
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Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Colorimetría , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética , ElectroquímicaRESUMEN
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.
Asunto(s)
Diospyros , Peróxido de Hidrógeno , Humanos , Peróxido de Hidrógeno/química , Diospyros/metabolismo , Colorimetría , Taninos , Citocromo P-450 CYP2B1 , Peroxidasa/químicaRESUMEN
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.
RESUMEN
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.
Asunto(s)
Técnicas Biosensibles , Glipicanos , Grafito , Nanopartículas del Metal , Humanos , Técnicas Biosensibles/métodos , Carcinoma Hepatocelular , Técnicas Electroquímicas/métodos , Grafito/química , Hemina/química , Peróxido de Hidrógeno , Neoplasias Hepáticas , Nanopartículas del Metal/química , Paladio , Plata/químicaRESUMEN
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.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Carcinoma Hepatocelular , Grafito , Neoplasias Hepáticas , Puntos Cuánticos , Humanos , Puntos Cuánticos/química , Grafito/química , Molibdeno/química , Nitrógeno/química , Neoplasias Hepáticas/diagnóstico , Óxidos de Nitrógeno , Aptámeros de Nucleótidos/química , Óxido Nítrico , Técnicas Biosensibles/métodosRESUMEN
The sensitivity and specificity of Golgi glycoprotein 73 (GP73) are very important for early diagnosis of hepatocellular carcinoma. Herein, we constructed a new-fashioned fluorescent aptamer sensor for GP73 determination based on nitrogen-doped graphene quantum dots (N-GQDS) and molybdenum disulfide (MoS2) nanosheets. N-GQDs with high fluorescence intensity and good stability were screened out, and GP73 aptamer (GP73Apt) is labeled with N-GQDs to form the N-GQDs-GP73Apt fluorescence probe. MoS2 nanosheets can quench the fluorescence of N-GQDs-GP73Apt owing to fluorescence resonance energy transfer mechanisms. After introducing GP73 into the biosensing system, the N-GQDs-GP73Apt specifically bound with GP73 to form the deployable structures, making N-GQDs-GP73Apt far away from MoS2, blocking the fluorescence energy transfer process, and restoring the fluorescence of N-GQDs-GP73Apt. When the GP73 concentration was in the extent of 2.5 ng/mLâ¼100 ng/mL, the relative fluorescence recovery is linearly relevant to the concentration of GP73, and the limit of detection (LOD) was 1.29 ng/mL (S/N = 3). Moreover in the application of actual serum sample detection, the recovery was range 98.85â¼100.55 %. The fluorescent aptamer sensor can rapidly detect and analyze the serum marker GP73 with the characteristics of low-cost, high sensitivity, good specificity and recovery.
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Aptámeros de Nucleótidos , Grafito , Puntos Cuánticos , Puntos Cuánticos/química , Molibdeno/química , Grafito/química , Nitrógeno/química , Aptámeros de Nucleótidos/químicaRESUMEN
Selective recovery of indium has been widely studied to improve the resource efficiency of critical metals. However, the interaction and selective adsorption mechanism of indium/iron ions with tannin-based adsorbents is still unclear and hinders further optimization of their selective adsorption performance. In this study, the epigallocatechin gallate (EGCG) monomer, which is the key functional unit of persimmon tannin, was chosen to explore the ability and mechanism of selective separation/extraction of indium from indium-iron mixture solutions. The density functional theory calculation results indicated that the deprotonated EGCG was easier to combine with indium/iron cations than those of un-deprotonated EGCG. Moreover, the interaction of the EGCG-Fe(III) complex was dominated by chelation and electrostatic interaction, while that of the EGCG-In(III) complex was controlled by electrostatic interactions and aromatic ring stacking effects. Furthermore, the calculation of binding energy verified that EGCG exhibited a stronger affinity for Fe(III) than that for In(III) and preferentially adsorbed iron ions in acidic or neutral solutions. Further experimental results were consistent with the theoretical study, which showed that the Freundlich equilibrium isotherm fit the In(III) and Fe(III) adsorption behavior very well, and the Fe(III) adsorption processes followed a pseudo-second-order model. Thermodynamics data revealed that the adsorption of In(III) and Fe(III) onto EGCG was feasible, spontaneous, and endothermic. The adsorption rate of the EGCG monomer for Fe(III) in neutral solution (1:1 mixed solution, pH = 3.0) was 45.7%, 4.3 times that of In(III) (10.7%). This study provides an in-depth understanding of the relationship between the structure of EGCG and the selective adsorption capacity at the molecular level and provides theoretical guidance for further optimization of the selective adsorption performance of structurally similar tannin-based adsorbents.
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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.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Carcinoma Hepatocelular , Grafito , Neoplasias Hepáticas , Nanopartículas del Metal , Humanos , Platino (Metal)/química , Paladio/química , Hemina , Nanopartículas del Metal/química , Técnicas Electroquímicas/métodos , Glipicanos , Grafito/química , Técnicas Biosensibles/métodos , Aptámeros de Nucleótidos/químicaRESUMEN
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.
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Técnicas Biosensibles , Quitosano , Nanopartículas del Metal , Quitosano/química , Colorimetría , Dimaprit/análogos & derivados , Grafito , Hemina , Humanos , Peróxido de Hidrógeno/química , Nanopartículas del Metal/química , Paladio/química , Peroxidasa/química , Peroxidasas , Platino (Metal)/químicaRESUMEN
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.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Carcinoma Hepatocelular , Grafito , Neoplasias Hepáticas , Nanopartículas del Metal , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Carcinoma Hepatocelular/diagnóstico , Técnicas Electroquímicas/métodos , Oro/química , Grafito/química , Hemina/química , Humanos , Peróxido de Hidrógeno/química , Límite de Detección , Neoplasias Hepáticas/diagnóstico , Nanopartículas del Metal/químicaRESUMEN
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.
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Técnicas Biosensibles , Quitosano , Grafito , Técnicas Biosensibles/métodos , Citocromo P-450 CYP2B1 , Desoxiglucosa , Técnicas Electroquímicas/métodos , Hemina , Humanos , Peróxido de Hidrógeno , Límite de Detección , Platino (Metal) , Reproducibilidad de los ResultadosRESUMEN
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.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Carcinoma Hepatocelular , Grafito , Neoplasias Hepáticas , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Carbono/química , Carcinoma Hepatocelular/diagnóstico , Detección Precoz del Cáncer , Transferencia Resonante de Energía de Fluorescencia/métodos , Glipicanos , Oro/química , Grafito/química , Humanos , Límite de Detección , Neoplasias Hepáticas/diagnósticoRESUMEN
Glypican-3 (GPC3), a heparin sulfate proteoglycan, is a potential diagnostic and therapeutic target for hepatocellular carcinoma. In this paper, a novel fluorescent aptasensor for GPC3 detection is constructed via glutathione@graphene quantum dots-labeled GPC3 aptamer (GSH@GQDs-GPC3Apt) as a fluorescence probe. First, GSH@GQDs is screened out with higher fluorescence intensity, which emits bright blue fluorescence under ultraviolet light. Then, the fluorescence-labeled GSH@GQDs-GPC3Apt probe is formed by the combination of amination GPC3Apt and GSH@GQDs using EDC/NHS coupled reaction. Under hydrogen bond and π-π interaction/stacking, the fluorescence of GSH@GQDs-GPC3Apt could be quenched by reductive graphene oxide (RGO) with the help of the photoinduced electron transfer and the fluorescence resonance energy transfer mechanism. In the presence of GPC3, the GSH@GQDs-GPC3Apt specifically recognizes and binds to GPC3, giving rise to the change of secondary structure of GPC3Apt to form the GPC3/GPC3Apt-GSH@GQDs complex, which would lead to the disintegration of the GSH@GQDs-GPC3Apt-RGO compound. Therefore, the energy transfer process is blocked and the fluorescence intensity is restored, enabling a highly sensitive response to GPC3. When the concentration of GPC3 is from 5.0â¯ng/mL to 150.0â¯ng/mL, the fluorescence recovery rate is well linearly related to GPC3 concentration with the limit of detection of 2.395â¯ng/mL (S/Nâ¯=â¯3). This strategy shows recoveries from 98.31% to 101.89% in human serum samples and provides simple, fast and cheap analysis of GPC3, which suggests that it has great potential applications in clinical diagnosis for hepatocellular carcinoma.
Asunto(s)
Grafito , Puntos Cuánticos , Glutatión , Glipicanos , Humanos , Límite de DetecciónRESUMEN
Antioxidants are molecules that can prevent the harmful effects of oxygen, help capture and neutralize free radicals, and thus eliminate the damage of free radicals to the human body. Persimmon tannin (PT) has excellent antioxidant activity, which is closely related to its molecular structure. We report here a comparative study of four characteristic structural units from PT (epicatechin gallate (ECG), epigallocatechin gallate (EGCG), A-type linked ECG dimer (A-ECG dimer), A-type linked EGCG dimer (A-EGCG dimer)) to explore the structure-activity relationship by using the density functional theory. Based on the antioxidation mechanism of hydrogen atom transfer, the most favorable active site for each molecule exerts antioxidant activity is determined. The structural parameters, molecular electrostatic potential, and frontier molecular orbital indicate that the key active sites are located on the phenolic hydroxyl group of the B ring for ECG and EGCG monomers, and the key active sites of the two dimers are located on the phenolic hydroxyl groups of the A and D' rings. The natural bond orbital and bond dissociation energy of the phenolic hydroxyl hydrogen atom show that the C11-OH in the ECG monomer and the C12-OH in the EGCG monomer are the most preferential sites, respectively. The most active site of the two A-linked dimers is likely located on the D' ring C20' phenolic hydroxyl group. Based on computational analysis of quantum chemical parameters, the A-ECG dimer is a more potent antioxidant than the A-EGCG dimer, ECG, and EGCG. This computational analysis provides the structure-activity relationship of the four characteristic units which will contribute to the development of the application of PT antioxidants in the future.
RESUMEN
A persimmon tannin-Aloe vera composite powder (PT-A) was investigated for its capacity to protect against ionizing radiation. Human hepatic cells (L02 cells) and human hepatoma cells (HepG2 cells) were pretreated with different concentrations of PT-A or the single compounds (PT or Aloe vera) and radiated with X-rays. After radiation and post-incubation for 12 h or 24 h, the cell viability, apoptosis, and reactive oxygen species (ROS) production were analyzed by Cell Counting Kit 8 (CCK-8), 2',7'-dichlorfluorescein diacetate (DCFH-DA) staining, and Hoechst 33258 staining/flow cytometry, respectively. CCK-8 results illustrated that the optimal radiation dose L02 cells was 8 Gy for L02 cells, and the cell activity was 71.72% (IC50 = 412.1 µg/mL) after post-radiation incubation of 12 h. For HepG2 cells, the optimal radiation dose was 8 Gy, and the cell activity was 62.37% (IC50 = 213.0 µg/mL). The cell apoptotic rate was the lowest at a PT-A concentration of 200 µg/mL in L02 cells (4.32%, P < 0.05), and at 100 µg/mL in HepG2 cells (9.80%, P < 0.05). ROS production induced by radiation could be effectively inhibited by 200 µg/mL of PT-A in L02 cells, and by 100 µg/mL of PT-A in HepG2 cells. The PT-A composite has good radioprotective effects on cell vitality and apoptosis of X-rays radiation exposure towards L02 cells and HepG2 cells compared to the persimmon tannin or Aloe vera. Therefore, PT-A composite might be useful as a natural, harmless anti-ionizing radiation agent, and has various clinical application prospects in future.