Hybrid carbon based nanomaterials for electrochemical detection of biomolecules

By combining different allotropic forms of carbon at the nanoscale it is possible to fabricate tailor made surfaces with unique properties. These novel materials have shown high potential especially in the electrochemical detection of different biomolecules, such as dopamine, glutamate and ascorbic acid, which are important neurotransmitters in the mammalian central nervous system. Thus, more information about their material properties must be obtained in order to realize their high potential to the maximum. The results presented in this review clearly point out that although there is an extensive amount of data available on the structural, chemical and electrochemical properties on different carbon nanoforms, the data are scattered, often inconsistent and even contradictory. Hybrid carbon nanomaterials are much less investigated than the individual allotropes, but based on the existing data they possess extremely interesting electrochemical properties. Thus, it is of utmost importance to carry out extensive step-by-step characterization of these materials by utilizing combination of detailed computational and experimental work. In this way it will become possible to avoid approaches to material design that are based solely on trial-and-error approach, which has, unfortunately, been more a rule than an exception.     

Metal nanoparticles as labels for heterogeneous, chip-based DNA detection

The last decade has witnessed the development of a variety of metal nanoparticle-based techniques for DNA detection. High sensitivity and specificity, miniaturization, and cost-efficient detection are problems addressed by the use of nanoparticle labels in heterogeneous DNA detection schemes. The small label size, established bioconjugation chemistry, and the unusual optical and electrical properties of metal nanoparticles make them unique tools for DNA detection. This paper reviews the different physical characteristics of metal nanoparticles and their implementation in assays. It covers various optical as well as gravimetric, electrochemical and electrical methods for analysing nanoparticle-labelled analytes, and particularly DNA, at sensing surfaces.     

In situ amplified electrochemical immunoassay for carcinoembryonic antigen using horseradish peroxidase-encapsulated nanogold hollow microspheres as labels

Methods based on sandwich-type electrochemical enzyme immunoassay protocol have been extensively developed for the detection of biomolecules, but most often exhibit low detection signals and low detection sensitivity, and are unsuitable for routine use. In this study, we initially synthesized specially horseradish peroxidase-encapsulated nanogold hollow microspheres (HRP-GHS), and then the prepared HRP-GHS was conjugated to the secondary carcinoembryonic antibody (HRP-GHS- anti-CEA). Carcinoembryonic antigen (CEA), as a model protein, was monitored by using the electrochemical sandwich-type enzyme immunoassay format. Under optimized conditions, the linear range of the immunoassay by using single HRP-labeled anti-CEA (HRP- anti-CEA) as secondary antibodies is 2.5-120 ng/mL with a detection limit of 1.5 ng/mL CEA, while the assay sensitivity by using HRP-GHS- anti-CEA as secondary antibodies is further increased from 0.01 to 200 ng/mL with a lower detection limit of 1.5 pg/mL CEA. The intra- and interassay reproducibility is acceptable. The CEA concentrations of the clinical serum specimens assayed by the developed immunoassay show consistent results in comparison with those obtained by commercially available enzyme-linked immunosorbent assay. This immunoassay system has many desirable merits including sensitivity, accuracy, and little required instrumentation. Significantly, the new protocol may be quite promising, with potentially broad applications for clinical immunoassays.     

Gold and silica-coated gold nanoparticles as thermographic labels for DNA detection

The infrared emissivity of Au and silica-coated Au nanoparticles (Au NPs) deposited on indium tin oxide substrates was investigated. NPs were irradiated with laser light at a frequency close to the Au plasmon resonance band, and the blackbody radiation emitted as a result was monitored with an IR camera equipped with an InAs array detector. The differences in temperature before and after laser irradiation were recorded (T-jumps) and were found to be directly proportional to the number of particles present on the slide and to the laser power used in the experiment. Coating Au NPs with silica increased the measured T-jumps 2-5 times, depending on the thickness of the silica shell. This was in agreement with the observation that silica has a much higher IR emissivity than Au. Both Au and silica-coated Au NPs were then tested as labels for thermographic DNA detection. Target DNA concentrations as low as 100 pM were recorded when Au NPs were used as labels and as low as 10 pM when silica-coated Au NPs were used.     

Tailored magnetic nanoparticles for direct and sensitive detection of biomolecules in biological samples

We developed nanoparticles with tailored magnetic properties for direct and sensitive detection of biomolecules in biological samples in a single step. Thermally blocked nanoparticles obtained by thermal hydrolysis, functionalized with specific ligands, are mixed with sample solutions, and the variation of the magnetic relaxation due to surface binding is used to detect the presence of biomolecules. The binding significantly increases the hydrodynamic volume of nanoparticles, thus changing their Brownian relaxation frequency which is measured by a specifically developed AC susceptometer. The system was tested for the presence of Brucella antibodies, a dangerous pathogen causing brucellosis with severe effects both on humans and animals, in serum samples from infected cows and the surface of the nanoparticles was functionalized with lipopolysaccharides (LPS) from Brucella abortus. The hydrodynamic volume of LPS-functionalized particles increased by 25-35% as a result of the binding of the antibodies, measured by changes in the susceptibility in an alternating magnetic field. The method has shown high sensitivity, with detection limit of 0.05 microg x mL(-1) of antibody in the biological samples without any pretreatment. This magnetic-based assay is very sensitive, cost-efficient, and versatile, giving a direct indication whether the animal is infected or not, making it suitable for point-of-care applications. The functionalization of tailored magnetic nanoparticles can be modified to suit numerous homogeneous assays for a wide range of applications.     

Metallophthalocyanine-nanofibre-based electrodes for electrochemical sensing of biomolecules

Metal phthalocyanines, possessing rich redox chemistry due to the presence of the central metal cation and pyrrolic nitrogen atoms of the macrocycle, are explored as electrochemical sensors. Nickel phthalocyanine nanofibres (NiPc NF) prepared by a simple chemical route are coated on a pencil graphite rod and the electrocatalytic performance of NiPc NF electrode is investigated for quantitative detection of ascorbic acid (AA) in 0.2 M phosphate buffer solution. The performance of NiPc NFs is shown to be superior to that of commercial NiPc and is attributed to the high electrochemically active surface area available for fibres. The electrode exhibits linearity for the detection over a wide concentration range of AA from \(5.5\,\upmu \hbox {M}\) to 5.2 mM. The detection limit for AA sensing with NiPc-NF-modified electrode is \(1.5\,\upmu \hbox {M}\). The higher performance of NiPc fibres due to its nanostructure morphology may be utilized for the quantitative detection of other biomolecules.     

Magnetic electrochemical immunoassays with quantum dot labels for detection of phosphorylated acetylcholinesterase in plasma

A new magnetic electrochemical immunoassay has been developed as a tool for biomonitoring exposures to organophosphate (OP) compounds, e.g., insecticides and chemical nerve agents, by directly detecting organophosphorylated acetylcholinesterase (OP-AChE). This immunoassay uniquely incorporates highly efficient magnetic separation with ultrasensitive square wave voltammetry (SWV) analysis with quantum dots (QDs) as labels. A pair of antibodies was used to achieve the specific recognition of OP-AChE that was prepared with paraoxon as an OP model agent. Antiphosphoserine polyclonal antibodies were anchored on amorphous magnetic particles preferably chosen to capture OP-AChE from the sample matrixes by binding their phosphoserine moieties that were exposed through unfolding the protein adducts. This was validated by electrochemical examinations and enzyme-linked immunosorbent assays. Furthermore, antihuman AChE monoclonal antibodies were labeled with cadmium-source QDs to selectively recognize the captured OP-AChE, as characterized by transmission electron microscopy. The subsequent electrochemical SWV analysis of the cadmium component released by acid from the coupled QDs was conducted on disposable screen-printed electrodes. Experimental results indicated that the SWV-based immunoassays could yield a linear response over a broad concentration range of 0.3-300 ng/mL OP-AChE in human plasma with a detection limit of 0.15 ng/mL. Such a novel electrochemical immunoassay holds great promise as a simple, selective, sensitive, and field-deployable tool for the effective biomonitoring and diagnosis of potential exposures to nerve agents and pesticides.     

SERS detection of biomolecules using lithographed nanoparticles towards a reproducible SERS biosensor

In this paper we highlight the accurate spectral detection of bovine serum albumin and ribonuclease-A using a surface-enhanced Raman scattering (SERS) substrate based on gold nanocylinders obtained by electron-beam lithography (EBL). The nanocylinders have diameters from 100 to 180 nm with a gap of 200 nm. We demonstrate that optimizing the size and the shape of the lithographed gold nanocylinders, we can obtain SERS spectra of proteins at low concentration. This SERS study enabled us to estimate high enhancement factors (10(5) for BSA and 10(7) for RNase-A) of important bands in the protein Raman spectrum measured for 1 mM concentration. We demonstrate that, to reach the highest enhancement, it is necessary to optimize the SERS signal and that the main parameter of optimization is the LSPR position. The LSPR have to be suitably located between the laser excitation wavelength, which is 632.8 nm, and the position of the considered Raman band. Our study underlines the efficiency of gold nanocylinder arrays in the spectral detection of proteins.     

Impedimetric biosensor based on magnetic nanoparticles for electrochemical detection of dopamine

One of the difficulties which limit the use of electrochemical sensors for detection of dopamine is the interference from ascorbic acid. We have sought to address this problem through the synthesis and characterization of a suitable electrode material based on magnetic nanoparticles. The interference from the ascorbic acid was overcome by fabricating a negatively charged electrode surface using PEGylated arginine functionalized magnetic nanoparticles (PA-MNPs). The nanoparticles were characterized by various techniques viz., X-ray diffraction, FT-Infrared spectroscopy, transmission electron microscopy and vibrating sample magnetometer. The electrochemical behavior of the proposed sensor was investigated by cyclic voltammetry and the sensor showed high sensitivity and selectivity for dopamine. The response mechanism of the modified electrode is based on the interaction between the negatively charged electrode and the positively charged dopamine. Under optimized conditions, linear calibration plots were obtained for amperometric detection of dopamine (DA) over the concentration range of 1–9 mM dopamine, with a linear correlation coefficient of 0.9836, sensitivity of 121 μA/mM and a detection limit of 7.25 μM. Electrochemical impedance spectroscopy (EIS) has been used to study the interface properties of modified electrodes. The value of the polarization resistance (R_p) increases linearly with dopamine concentration in the range of 10 μM to 1 mM and the limit of detection (LOD) was calculated to be 14.1 μM. High sensitivity and selectivity, micromolar detection limit, high reproducibility, along with ease of preparation of the electrode surface make this system suitable for the determination of DA in pharmaceutical and clinical preparations.     

DNA-wrapped carbon nanotubes as sensitive electrochemical labels in controlled-assembly-mediated signal transduction for the detection of sequence-specific DNA

A novel electrochemical strategy that uses DNA-wrapped carbon nanotubes (CNTs) as electrochemical labels is developed for sensitive and selective detection of sequence-specific DNA. The presence of target DNA mediates the formation of a sandwiched complex between the DNA-wrapped CNT and a hairpin DNA capture probe immobilized on magnetic beads. This allows target-selective collection of the CNT labels by magnetic separation and transfer on the electrode surface modified with an insulating self-assembled monolayer (SAM). After treatment with N,N-dimethylformamide, the collected sandwiched complex releases the bare CNTs and facilitates the removal of magnetic beads from the electrode surface. The bare CNTs can then assemble on the SAM-modified electrode surface and mediate efficient electron transfer between the electrode and the electroactive species in the solution with a strong current signal generated. The results indicate that the developed strategy shows a sensitive response to target DNA with a desirable signal gain and a low detection limit of 0.9 pM. This strategy is also demonstrated to provide excellent differentiation of single-base mismatch in target DNA. It is expected that this electrochemical strategy may hold great potential as a novel platform for clinical diagnostics and genetic analysis.     

Silicon nanoribbons for electrical detection of biomolecules

Direct electrical detection of biomolecules at high sensitivity has recently been demonstrated using semiconductor nanowires. Here we demonstrate that semiconductor nanoribbons, in this case, a thin sheet of silicon on an oxidized silicon substrate, can approach the same sensitivity extending below the picomolar concentration regime in the biotin/streptavidin case. This corresponds to less than approximately 20 analyte molecules bound to receptors on the nanoribbon surface. The micrometer-size lateral dimensions of the nanoribbon enable optical lithography to be used, resulting in a simple and high-yield fabrication process. Electrical characterization of the nanoribbons is complemented by computer simulations showing enhanced sensitivity for thin ribbons. Finally, we demonstrate that the device can be operated both in inversion as well as in accumulation mode and the measured differences in detection sensitivity are explained in terms of the distance between the channel and the receptor coated surface with respect to the Debye screening length. The nanoribbon approach opens up for large scale CMOS fabrication of highly sensitive biomolecule sensor chips for potential use in medicine and biotechnology.     

The simple preparation of graphene/Pt nanoparticles composites and their electrochemical performance

The graphene-Pt nanoparticles (NPs) composites were prepared with ascorbic acid as the reductant through in-situ reduction method. The graphene-Pt NPs were characterized by X-ray diffraction, the Fourier-transform infrared spectra, Raman spectroscopy, scanning electron microscopy and transmission electron microscopy. The obtained production was loaded with Pt nanoparticles, which could increase the active surface area of the electrode catalyst and accelerated the electron transfer.     

Coulometric detection of irreversible electrochemical reactions occurring at Pt microelectrodes used for neural stimulation

The electrochemistry of 50 μm diameter Pt electrodes used for neural stimulation was studied in vitro by reciprocal derivative chronopotentiometry. This differential method provides well-defined electrochemical signatures of the various polarization phenomena that occur at Pt microelectrodes and are generally obscured in voltage transients. In combination with a novel in situ coulometric approach, irreversible H(2) and O(2) evolution, Pt dissolution and reduction of dissolved O(2) were detected. Measurements were performed with biphasic, charge-balanced, cathodic-first and anodic-first current pulses at charge densities ranging from 0.07 to 1.41 mC/cm(2) (real surface area) in phosphate buffered saline (PBS) with and without bovine serum albumin (BSA). The extent to which O(2) reduction occurs under the different stimulation conditions was compared in O(2)-saturated and deoxygenated PBS. Adsorption of BSA inhibited Pt dissolution as well as Pt oxidation and oxide reduction by blocking reactive sites on the electrode surface. This inhibitory effect promoted the onset of irreversible H(2) and O(2) evolution, which occurred at lower charge densities than those in PBS. Reduction of dissolved O(2) on Pt electrodes accounted for 19-34% of the total injected charge in O(2)-saturated PBS, while a contribution of 0.4-12% was estimated for in vivo stimulation. These result may prove important for the interpretation of histological damage induced by neural stimulation and therefore help define safer operational limits.     

Catalytic oxidation of carbon monoxide over radiolytically prepared Pt nanoparticles supported on glass

Platinum nanoparticles have been prepared by radiolytic and chemical methods in the presence of stabilizer gelatin and SiO₂ nanoparticles. The formation of Pt nanoparticles was confirmed using UV-vis absorption spectroscopy and transmission electron microscopy (TEM). The prepared particles were coated on the inner walls of the tubular pyrex reactor and tested for their catalytic activity for oxidation of CO. It was observed that Pt nanoparticles prepared in the presence of a stabilizer (gelatin) showed a higher tendency to adhere to the inner walls of the pyrex reactor as compared to that prepared in the presence of silica nanoparticles. The catalyst was found to be active at ≥150 °C giving CO₂. Chemically reduced Pt nanoparticles stabilized on silica nanoparticles gave ∼7% CO conversion per hour. However, radiolytically prepared Pt nanoparticles stabilized by gelatin gave ∼10% conversion per hour. Catalytic activity of radiolytically prepared platinum catalyst, coated on the inner walls of the reactor, was evaluated as a function of CO concentration and reaction temperature. The rate of reaction increased with increase in reaction temperature and the activation energy for the reaction was found to be ∼108.8 kJ mol⁻¹. The rate of CO₂ formation was almost constant (∼1.5 × 10⁻⁴ mol dm⁻³ h⁻¹) at constant O₂ concentration (6.5 × 10⁻³ mol dm⁻³) with increase in CO concentration from 2 × 10⁻⁴ mol dm⁻³ to 3.25 × 10⁻³ mol dm⁻³. The data indicate that catalytic oxidation of CO takes place by Eley-Rideal mechanism.     

Magnetic nanoparticles applied in electrochemical detection of controllable DNA hybridization

Electrochemical detection of hybridized DNA strands was achieved with a magnetic nanoparticle modified electrode and the commonly used electrochemical couple K3[Fe(CN)6]/K4[Fe(CN)6]. The detection proved to be fast and very simple. Furthermore, magnetic nanoparticles could be employed to control the DNA hybridization process. An inhibited or an enhanced degree of hybridizing could be produced.     

An electrochemical sensor based on V2O5 nanoparticles for the detection of ciprofloxacin

This research constructed and utilized a screen-printed electrode (SPE) modified with V₂O₅ nanoparticles (NPs) (V₂O₅/SPE) employed in order to sensitively and selectively quantify ciprofloxacin with exceptional accuracy in the phosphate buffer solution (PBS) with the use of differential pulse voltammetry (DPV). Moreover, electrochemical properties of this new V₂O₅/SPE sensor have been examined using diverse characterization procedures. Very good V₂O₅/SPE electrochemical features offered sensitive ciprofloxacin voltammetric determination with the reduced limit of detection (LOD?=?0.01 µM) toward electrocatalytic oxidation of ciprofloxacin in comparison with the bare SPE. Finally, this new disposable sensor exhibited higher sensitivity and thus has been efficiently utilized to determine ciprofloxacin in the real samples.