Glucose oxidase-catalyzed growth of gold nanoparticles enables quantitative detection of attomolar cancer biomarkers.
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ABSTRACT:
SUBMITTER: Liu D
PROVIDER: S-EPMC4066917 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:We present here an ultrasensitive electrochemical biosensor based on a lectin biorecognition capable to detect concentrations of glycoproteins down to attomolar (aM) level by investigation of changes in the charge transfer resistance (Rct) using electrochemical impedance spectroscopy (EIS). On polycrystalline gold modified by an aminoalkanethiol linker layer, gold nanoparticles were attached. A Sambucus nigra agglutinin was covalently immobilised on a mixed self-assembled monolayer formed on gold nanoparticles and finally, the biosensor surface was blocked by poly(vinyl alcohol). The lectin biosensor was applied for detection of sialic acid containing glycoproteins fetuin and asialofetuin. Building of a biosensing interface was carefully characterised by a broad range of techniques such as electrochemistry, EIS, atomic force microscopy, scanning electron microscopy and surface plasmon resonance with the best performance of the biosensor achieved by application of HS-(CH2)11-NH2 linker and gold nanoparticles with a diameter of 20 nm. The lectin biosensor responded to an addition of fetuin (8.7% of sialic acid) with sensitivity of (338 ± 11) ? decade(-1) and to asialofetuin (? 0.5% of sialic acid) with sensitivity of (109 ± 10) ? decade(-1) with a blank experiment with oxidised asialofetuin (without recognisable sialic acid) revealing sensitivity of detection of (79 ± 13) ? decade(-1). These results suggest the lectin biosensor responded to changes in the glycan amount in a quantitative way with a successful validation by a lectin microarray. Such a biosensor device has a great potential to be employed in early biomedical diagnostics of diseases such as arthritis or cancer, which are connected to aberrant glycosylation of protein biomarkers in biological fluids.
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Project description:Biosensors for the determination of glucose concentration have a great significance in clinical diagnosis, and in the food and pharmaceutics industries. In this research, short-chain polyaniline (PANI) and polypyrrole (Ppy)-based nanocomposites with glucose oxidase (GOx) and 6 nm diameter AuNPs (AuNPs(6 nm)) were deposited on the graphite rod (GR) electrode followed by the immobilization of GOx. Optimal conditions for the modification of GR electrodes by conducting polymer-based nanocomposites and GOx were elaborated. The electrodes were investigated by cyclic voltammetry and constant potential amperometry in the presence of the redox mediator phenazine methosulfate (PMS). The improved enzymatic biosensors based on GR/PANI-AuNPs(6 nm)-GOx/GOx and GR/Ppy-AuNPs(6 nm)-GOx/GOx electrodes were characterized by high sensitivity (65.4 and 55.4 μA mM-1 cm-2), low limit of detection (0.070 and 0.071 mmol L-1), wide linear range (up to 16.5 mmol L-1), good repeatability (RSD 4.67 and 5.89%), and appropriate stability (half-life period (τ1/2) was 22 and 17 days, respectively). The excellent anti-interference ability to ascorbic and uric acids and successful practical application for glucose determination in serum samples was presented for GR/PANI-AuNPs(6 nm)-GOx/GOx electrode.
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Project description:We report the microfabrication and characterization of gold microband electrodes on silicon using standard microfabrication methods, i.e., lithography and etching techniques. A two-step electrodeposition process was carried out using the on-chip platinum reference and gold counter electrodes, thus incorporating glucose oxidase onto a platinum-modified, gold microband electrode with an o-phenylenediamine and ß-cyclodextrin mixture. The as-fabricated electrodes were studied using optical microscopy, scanning electron microscopy, and atomic force microscopy. The two-step electrodeposition process was conducted in low sample volumes (50 µL) of both solutions required for biosensor construction. Cyclic voltammetry and electrochemical impedance spectroscopy were utilised for electrochemical characterization at each stage of the deposition process. The enzymatic-based microband biosensor demonstrated a linear response to glucose from 2.5-15 mM, using both linear sweep voltammetry and chronoamperometric measurements in buffer-based solutions. The biosensor performance was examined in 30 µL volumes of fetal bovine serum. Whilst a reduction in the sensor sensitivity was evident within 100% serum samples (compared to buffer media), the sensor demonstrated linear glucose detection with increasing glucose concentrations (5-17 mM).
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Project description:Nanozymes, defined as nanomaterials that can mimic the catalytic activity of natural enzymes, have been widely used to develop analytical tools for biosensing. In this regard, the monitoring of glutathione (GSH), a key antioxidant biomolecule intervening in the regulation of the oxidative stress level of cells or related with Parkinson's or mitochondrial diseases can be of great interest from the biomedical point of view. In this work, we have synthetized a gold-platinum Au@Pt nanoparticle with core-shell configuration exhibiting a remarkable oxidase-like mimicking activity towards the substrates 3,3',5,5'-tetramethylbenzidine (TMB) and o-phenylenediamine (OPD). The presence of a thiol group (-SH) in the chemical structure of GSH can bind to the Au@Pt nanozyme surface to hamper the activation of O2 and reducing its oxidase-like activity as a function of the concentration of GSH. Herein, we exploit the loss of activity to develop an analytical methodology able to detect and quantify GSH up to µM levels. The system composed by Au@Pt and TMB demonstrates a good linear range between 0.1-1.0 µM to detect GSH levels with a limit of detection (LoD) of 34 nM.
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Project description:Glucose oxidase (GOx)-based electrodes are important for bioelectronics, such as glucose sensors. It is challenging to effectively link GOx with nanomaterial-modified electrodes while preserving enzyme activity in a biocompatible environment. To date, no reports have used biocompatible food-based materials, such as egg white proteins, combined with GOx, redox molecules, and nanoparticles to create the biorecognition layer for biosensors and biofuel cells. This article demonstrates the interface of GOx integrated with egg white proteins on a 5 nm gold nanoparticle (AuNP) functionalized with a 1,4-naphthoquinone (NQ) and conjugated with a screen-printed flexible conductive carbon nanotube (CNT)-modified electrode. Egg white proteins containing ovalbumin can form three-dimensional scaffolds to accommodate immobilized enzymes and adjust the analytical performance. The structure of this biointerface prevents the escape of enzymes and provides a suitable microenvironment for the effective reaction. The bioelectrode's performance and kinetics were evaluated. Using redox-mediated molecules with the AuNPs and the three-dimensional matrix made of egg white proteins improves the transfer of electrons between the electrode and the redox center. By engineering the layer of egg white proteins on the GOx-NQ-AuNPs-mediated CNT-functionalized electrodes, we can modulate analytical performances such as sensitivity and linear range. The bioelectrodes demonstrate high sensitivity and can prolong the stability by more than 85% after 6 h of continuous operation. The use of food-based proteins with redox molecule-modified AuNPs and printed electrodes demonstrates advantages for biosensors and energy devices due to their small size, large surface area, and ease of modification. This concept holds a promise for creating biocompatible electrodes for biosensors and self-sustaining energy devices.
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Project description:This paper describes a low-cost facile method to construct gold (Au) nanoparticles (NPs) modified copper oxide (CuO) nanowires (NWs) electrode on copper foil for the detection of glucose. Copper foil has been converted to aligned CuO NWs arrays by sequential formation of Cu(OH)2 followed by heat treatment induced phase transformation to CuO. Au NPs are deposited on CuO NWs via simple reductive solution chemistry to impart high surface to volume ratio and enhanced catalytic activity of the resulting electrode. Structure, microstructure and morphology of Cu, Cu(OH)2 NWs, CuO NWs, and Au NPs modified CuO NWs are investigated by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The homogeneous distribution of Au NPs (average diameter ∼12 nm) on CuO NWs (average diameter 100 nm and aspect ratio ∼20) is confirmed by high-resolution transmission electron microscopy (HRTEM), scanning transmission electron microscopy (STEM) and elemental mapping. This CuO based glucose detection method gives the highest sensitivity along with the maximum linearity range. This non-enzymatic glucose sensor based on Au modified CuO NWs electrode gives broad linearity range from 0.5 μM to 5.9 mM. The sensor exhibits sensitivity of 4398.8 μA mM-1 cm-2, lower detection limit of 0.5 μM, and very fast response time of ∼5 s. Properties of the proposed glucose sensor are also investigated in human blood and it is found that the sensor is highly accurate and reliable. In addition, higher sensitivity and lower detection limit confirm that this device is suitable for invasive detection in saliva and urine.
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Project description:We report that entrapping glucose oxidase (GOx) within metallic gold, expands its activity to become an oxidase for monosaccharides that do not have a natural enzyme with that activity-fructose and xylose-and that this entrapment also removes the enantioselectivity, rendering this enzyme capable of oxidizing the "wrong" L-enantiomer of glucose. These observations suggest that in this biomaterial adsorptive interactions of the outer regions of the protein with the gold cage, pull apart and widen the tunnel between the two monomeric units of GOx, to a degree that its stereoselectivity is compromised; then, the active sites which are more versatile than currently attributed to, are free and capable of acting on the foreign sugars. To test this proposition, we entrapped in gold L-asparaginase, which is also a dimeric enzyme (a dimer of tight dimers), and found, again, that this metallic biomaterial widens the activity of that enzyme, to include the D-amino acid counter enantiomer as well. Detailed kinetic analyses for all substrates are provided for the gold bio-composites, including determination of the difference between the activation energies towards two opposite enantiomers.
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Project description:Using the previously designed biphenyl-2-ylphosphine ligand, featuring a remote tertiary amino group, the first gold-catalyzed intermolecular hydroalkenylation of alkynes has been developed. Synthetically valuable conjugated dienyl alcohols are formed in moderate to good yields. A range of alkenyltrifluoroborates are allowed as the alkenyl donor, and no erosion of alkene geometry and/or the propargylic configuration are detected. DFT calculations confirm the critical role of the remote basic group in the ligand as a general-base catalyst for promoting this novel gold catalysis with good efficiency.
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Project description:Recently, personal glucose meter (PGM) has been utilized for the detection of non-glucose targets for point-of-care (POC) testing. Aimed at this goal, we herein developed a new PGM-based label-free read-out method for polymerase chain reaction (PCR) based on our novel finding that cerium oxide nanoparticles (CeO2 NPs) exhibit glucose oxidase-like activity comparable to the natural glucose oxidase enzyme. Methods: In principle, DNA amplicons produced by PCR in the presence of target DNA electrostatically bind to CeO2 NPs, leading to their aggregation and reducing the efficiency for CeO2 NP-catalyzed glucose oxidation reaction. Thus, glucose is hardly oxidized to gluconic acid, resulting in the maintenance of initial high glucose level. On the contrary, in the absence of target DNA or presence of non-target DNA, DNA amplicons are not produced and glucose is effectively oxidized by the glucose oxidase-like activity of CeO2 NPs, leading to the significant reduction of glucose level. Finally, the resulting glucose level is simply measured by using PGM. Results: With this strategy, DNA amplicons were quantitatively examined within 5 min, realizing ultrafast analysis of PCR results without any cumbersome and labor-intensive procedures. In addition, the target genomic DNA derived from Escherichia coli (E. coli) was sensitively determined down to 10 copies with high selectivity. Conclusion: Importantly, the use of PGM as a detection component enables its direct application in POC settings. Based on the meritorious features of PGM such as rapidity, simplicity, and cost-effectiveness, we expect that the devised system could serve as a core platform for the on-site read-out of PCR amplification.
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Project description:In this work, we report the synthesis and biophysical studies carried out on a new kind of biocompatible and very stable gold nanoparticle (GNP) stabilized with glucose through a PEG linker (AuNP-PEG-Glu). The synthetic path was optimized to obtain nanoparticles of controlled sizes. ζ-potential and dynamic light scattering measurements allowed assessment of the nanodimension, dispersity, surface charge, and stability of our GNPs. Confocal microscopy demonstrated qualitatively that glucose molecules are successfully bonded to GNP surfaces. For our study, we selected nanoparticles with diameter in a range that maximizes the internalization efficiency in cells (40 nm). A detailed investigation about the biophysical proprieties of AuNP-PEG-Glu was carried out by means of fluorescence correlation spectroscopy (FCS) and orbital tracking techniques. This work gives new insights about the uptake mechanism of gold nanoparticles capped with glucose molecules.
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