Point-of-care detection of Microcystin-LR with a personal glucose meter in drinking water source

In this study, we described a point-of-care sensing protocol for rapid and sensitive detection of Microcystin-LR (MC-LR) in water by personal glucose meter. The POCT method possessed good reproducibility, selectivity, and stability, which may have potential for many other on-site detection applications.     

Rapid determination of the glucose content of molasses using a biosensor

A knowledge of the sugar content of molasses is of commercial importance to a number of industrial fermentations. Hence the feasibility of using a glucose oxidase biosensor to determine the glucose content of molasses samples was investigated. This method was compared with standard high-performance liquid chromatographic (HPLC) and gas-liquid chromatographic (GLC) procedures and with the use of a commercially available glucose analyser. A good correlation was obtained between the standard acetic anhydride GLC and glucose oxidase biosensor results (correlation coefficient = 0.98). Rapid and accurate measurements could be carried out using the biosensor without the need to employ the sample preparation step required in standard GLC methods. It was concluded that the use of the biosensor technique for the determination of glucose in molasses samples has distinct advantages over conventional methods.     

Quantum dot-based immunosensor for the detection of prostate-specific antigen using fluorescence microscopy

A sensitive optical method based on quantum dot (QD) technology is demonstrated for the detection of an important cancer marker, total prostate-specific antigen (TPSA) on a disposable carbon substrate surface. Immuno-recognition was carried out on a carbon substrate using a sandwich assay approach, where the primary antibody (Ab)-protein A complex covalently bound to the substrate surface, was allowed to capture TPSA. After the recognition event, the substrate was exposed to the biotinylated secondary Abs. After incubation with the QD streptavidin conjugates, QDs were captured on the substrate surface by the strong biotin-streptavidin affinity. Fluorescence imaging of the substrate surface illuminated the QDs, and provided a very sensitive tool for the detection of TPSA in undiluted human serum samples with a detection limit of 0.25 ng/mL. The potential of this method for application as a simple and efficient diagnostic strategy for immunoassays is discussed.     

Digital droplet immunoassay based on a microfluidic chip with magnetic beads for the detection of prostate-specific antigen

Sensitive biomarker detection techniques are beneficial for both disease diagnosis and postoperative examinations. In this study, we report an integrated microfluidic chip designed for the immunodetection of prostate-specific antigens (PSAs). The microfluidic chip is based on the three-dimensional structure of quartz capillaries. The outlet channel extends to 1.8 cm, effectively facilitating the generation of uniform droplets ranging in size from 3 to 50 μm. Furthermore, we successfully immobilized the captured antibodies onto the surface of magnetic beads using an activator, and we constructed an immunosandwich complex by employing biotinylated antibodies. A key feature of this microfluidic chip is its integration of microfluidic droplet technology advantages, such as high-throughput parallelism, enzymatic signal amplification, and small droplet size. This integration results in an exceptionally sensitive PSA detection capability, with the detection limit reduced to 7.00 ± 0.62 pg/mL.     

Ultrasensitive electrochemical detection of prostate-specific antigen (PSA) using gold-coated magnetic nanoparticles as 'dispersible electrodes'

Herein, we demonstrate the use of modified gold-coated magnetic nanoparticles as 'dispersible electrodes' which act as selective capture vehicles for electrochemical detection of prostate-specific antigen (PSA). A key advantage of this system is the ability to quantify non-electrochemical active analytes such as proteins with unprecedented detection limits and fast response times.     

Integrated biosensor for glucose and galactose

A glucose microsensor based on quinoprotein glucose dehydrogenase (GDH) and electron mediators is described. It is unaffected by the concentration of dissolved oxygen. Its calibration graph is linear below 10 mg dl ^?1. The insensitivity to oxygen arises because the concentration of the oxidized forms of the mediators is insufficient to oxidize the reduced form of GDH. An integrated microsensor for glucose and galactose based on GHD and galactose oxidase was constructed. Glucose and galactose concentrations were determined from the current increase due to oxidation of the mediators or the current decrease due to reduction of oxygen.     

A voltammetric immunoassay for the carcinoembryonic antigen using a self-assembled magnetic nanocomposite

The authors describe a voltammetric immunoassay for the carcinoembryonic antigen (CEA). It is based on the use of a self-assembled magnetic nanocomposite as multifunctional signal amplification platform. The core of the nanocomposite consists of Fe₃O₄ microspheres, and the shell of zirconium hexacyanoferrate loaded with gold nanoparticles (AuNPs@ZrHCF@Fe₃O₄). The material was synthesized by an electrostatic self-assembly process which is caused by the strong interaction between cyano groups and AuNPs. The surface of the Fe₃O₄ microspheres was functionalized with amino groups to facilitate the immobilization of ZrHCF which acts as an electron mediator. The nanocomposite was placed on a glassy carbon electrode which then displays noteworthy electrocatalytic activity toward the reduction of hydrogen peroxide (H₂O₂). The AuNPs serve as a support for the immobilization of antibodies by the interaction between AuNPs and amino groups on antibodies to construct a covalent Au-N bond. This facilitates electron transfer on the electrode surface using H₂O₂ as the electrochemical probe. Square wave voltammetry (measured typically at +0.2 V vs. SCE) was carried out to record the electrochemical behavior. Under the optimal conditions, a response is linear in the 0.5 pg·mL^?1 to 50 ng·mL^?1 CEA concentration range, and the detection limit is as low as 0.15 pg·mL^?1 (S/N =?3). The method is selective, highly stable and acceptably reproducible.

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Graphical abstract A self-assembly magnetic nanocomposite for voltammetric immunoassay of CEA. GCE glassy carbon electrode; Au NPs gold nanoparticles; ZrHCF zirconium hexacyanoferrate; CEA carcinoembryonic antigen; Anti-CEA CEA antibody; BSA bovine serum albumin; SWV square wave voltammetry. A high sensitive voltammetric immunoassay method has been used for detecting CEA, It is based on a self-assembled magnetic nanocomposite (Au NPs@ZrHCF@Fe₃O₄) as multifunctional signal amplification platform.

     

In-situ quantitative analysis of a prostate-specific antigen (PSA) using a nanomechanical PZT cantilever

We report on a novel technique of resonant frequency shift measurement based on a nanomechanical cantilever with a PZT actuating layer for label-free detection of a prostate-specific antigen (PSA) in a liquid environment. The nanomechanical PZT thin film cantilever is composed of SiO(2)/Ta/Pt/PZT/Pt/SiO(2) on a SiN(x) supporting layer for simultaneous self-exciting and sensing; it was fabricated using a standard MEMS (micro electromechanical system) process. The specific binding characteristics of the PSA antigen to its antibody, which is immobilized with Calixcrown self-assembled monolayers (SAMs) on a gold surface deposited on a cantilever, are determined to a high sensitivity. For the bioassay in a liquid environment, a liquid test cell with a 20 microl volume reaction chamber has been fabricated, using a bonding technique between poly(dimethyl siloxane) (PDMS) bilayers. An observed trend of resonant frequency change with respect to time could be explained by the binding kinetics due to the Langmuir isotherm and diffusion and by the effects of a small volume reaction chamber. In the saturated regimes, the resonant frequency of the cantilever increased with increase of the PSA concentration in the reaction chamber, showing that the trend of the resonance frequency change was similar to that of the fluorescence results. The saturated resonance frequency shift of the cantilever was proportional to the PSA antigen concentration of analyte solution.     

Sandwich immunoassay for the prostate specific antigen using a micro-fluxgate and magnetic bead labels

We describe a micro fluxgate based device with rectangular magnetic core for the determination of prostate specific antigen (PSA) labeled with Dynabeads. A sandwich immunoassay was employed where PSA is captured on a gold film modified with a self-assembled monolayer of antibody. The secondary antibody is labeled with Dynabeads. By applying a DC magnetic fields in the range of 460 to 700 μT, PSA can be detected with detection limit as low as 0.1 ng mL^?1. This micro fluxgate-based assay offers the advantages of miniaturization, simple and conveniently manipulation, re-usability and stability. In our perception, it offers a viable approach towards clinical determination of PSA or other biomarkers.

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Graphical abstract A separable detection method based on micro fluxgate and immunomagnetic beads was developed for detection of prostate specific antigen (PSA). Sandwich immunoassay was employed where PSA is captured on a gold film modified with a self-assembled monolayer of antibody. By applying a DC magnetic fields in the range of 460 to 700 μT, PSA can be detected with detection limit as low as 0.1 ng mL^?1, and this bio-sensing system can also give an approximate quantitation to the concentrations of them.

     

A novel method for glucose determination based on electrochemical impedance spectroscopy using glucose oxidase self-assembled biosensor

A method is developed for quantitative determination of glucose using electrochemical impedance spectroscopy (EIS). The method is based on immobilized glucose oxidase (GOx) on the topside of gold mercaptopropionic acid self-assembled monolayers (Au-MPA-GOx SAMs) electrode and mediation of electron transfer by parabenzoquinone (PBQ). The PBQ is reduced to hydroquinone (H(2)Q), which in turn is oxidized at Au electrode in diffusion layer. An increase in the glucose concentration results in an increase in the diffusion current density of the H(2)Q oxidation, which corresponds to a decrease in the faradaic charge transfer resistance (R(ct)) obtained from the EIS measurements. Glucose is quantified from linear variation of the sensor response (1/R(ct)) as a function of glucose concentration in solution. The method is straightforward and nondestructive. The dynamic range for determination of glucose is extended to more than two orders of magnitude. A detection limit of 15.6 microM with a sensitivity of 9.66 x 10(-7) Omega(-1)mM(-1) is obtained.     

A promising dehydrogenase-based bioanode for a glucose biosensor and glucose/O2 biofuel cell

A new type of dehydrogenase-based amperometric glucose biosensor was constructed using glucose dehydrogenase (GDH) which was immobilized on the edge-plane pyrolytic graphite (EPPG) electrode modified with poly(phenosafranin)-functionalized single-walled carbon nanotubes (PPS-SWCNTs). The PPS-SWCNT-modified EPPG electrode was prepared by electropolymerization of phenosafranin on the EPPG electrode which had been previously coated with SWCNTs. The performance of the GDH/PPS-SWCNT/EPPG bioanode was evaluated using cyclic voltammetry and amperometry in the presence of glucose. The GDH/PPS-SWCNT/EPPG electrode possesses promising characteristics as a glucose sensor: a wide linear dynamic range of 50 to 700 μM, low detection limit of 0.3 μM, fast response time (1-2 s), high sensitivity (96.5 μA cm(-2) mM(-1)), and anti-interference and anti-fouling abilities. Moreover, the performance of the GDH/PPS-SWCNT/EPPG bioanode was tested in a glucose/O(2) biofuel cell. The maximum power density delivered by the assembled glucose/O(2) biofuel cell could reach 64.0 μW cm(-2) at a cell voltage of 0.3 V with 40 mM glucose.     

Antibody-functionalized magnetic nanoparticles for the detection of carcinoembryonic antigen using a flow-injection electrochemical device

A magnetocontrolled immunosensing strategy based on flow-injection electrochemical impedance spectroscopy (EIS) was developed for the determination of carcinoembryonic antigen (CEA) in human serum. The immunosensor was fabricated by immobilizing anti-CEA on epoxysilane-modified core–shell magnetic Fe₃O₄/SiO₂ nanoparticles. The detection principle is based on the difference between the resistances measured before and after the antigen–antibody interaction. The performance of the immunosensor and factors influencing this performance were also proposed. The resistance response depended linearly on the CEA concentration over the range 1.5–60 ng/ml, and the immunosensor gave a detection limit of 0.5 ng/ml (S/N = 3). Coefficients of variance (CVs) of <9.8% were obtained for the intra- and interassay precisions. The method was successfully applied to the analysis of CEA in human serum. The recoveries obtained by spiking CEA standards into normal serum were 87–113%. The performance of the immunosensor was compared with a commercially available CEA ELISA. Satisfactory results were obtained according to a paired t-test method (t value < t _critical at the 95% confidence level). Importantly, the proposed immobilization protocol could be further developed to immobilize other antigens or biocompounds. Figure This study introduced a magnetocontrolled electrochemical immunosensing strategy based on antibody-functionalized magnetic core–shell Fe₃O₄/SiO₂ nanoparticles for the determination of carcinoembryonic antigen in human serum     

Complementary detection of prostate-specific antigen using In2O3 nanowires and carbon nanotubes

We report complementary detection of prostate-specific antigen (PSA) using n-type In2O3 nanowires and p-type carbon nanotubes. Our innovation involves developing an approach to covalently attach antibodies to In2O3 NW surfaces via the onsite surface synthesis of phosphonic acid-succinylimide ester. Electronic measurements under dry conditions revealed complementary response for In2O3 NW and SWNT devices after the binding of PSA. Real-time detection in solution has also been demonstrated for PSA down to 5 ng/mL, a benchmark concentration significant for clinical diagnosis of prostate cancer, which is the most frequently diagnosed cancer.     

Endonuclease-based logic gates and sensors using magnetic force-amplified readout of DNA scission on cantilevers

The endonuclease scission of magnetic particles functionalized with sequence-specific DNAs, which are associated on cantilevers, is followed by the magnetic force-amplified readout of the reactions by the nano-mechanical deflection/retraction of the cantilevers. The systems are employed to develop AND or OR logic gates and to detect single base mismatch specificity of the endonucleases. The two endonucleases EcoRI (E(A)) and AscI (E(B)) are used as inputs. The removal of magnetic particles linked to the cantilever by the duplexes 1/1a and 2/2a via the simultaneous cleavage of the DNAs by E(A) and E(B) leads to the retraction of the magnetically deflected cantilever and to the establishment of the "AND" gate. The removal of the magnetic particles linked to the cantilevers by the duplex 3/3a by either E(A) or E(B) leads to the retraction of the magnetically deflected cantilever and to the establishment of the "OR" gate. The magnetic force-amplified readout of endonuclease activities is also employed to reveal single base mismatch specificity of the biocatalysts.     

Rapid and simple preparation of a reagentless glucose electrochemical biosensor

A rapid and simple procedure was developed for the preparation of a highly stable and leach-proof glucose oxidase (GOx)-bound glassy carbon electrode (GCE). Crosslinked GOx via glutaraldehyde was drop-cast on a KOH-pretreated GCE followed by drop-casting of 3-aminopropyltriethoxysilane (APTES) to form a stable bioactive layer. At -0.45 V, the biosensor exhibited a wide dynamic detection range of 0.5-48 mM for commercial glucose and 1.3-28.2 mM for Sugar-Chex blood glucose linearity standards. Several endogenous electroactive substances and drug metabolites commonly found in blood were tested and provoked no signal response. To our knowledge, the developed procedure is the most rapid method for preparing a glucose biosensor. The biosensor suffered no biofouling after 7 days of immersion in Sugar-Chex blood glucose. With excellent production reproducibility, GOx-bound electrodes stored dry at room temperature retained their initial activity after several weeks.     

Disposable electrochemiluminescent biosensor using bidentate-chelated CdTe quantum dots as emitters for sensitive detection of glucose

A novel disposable solid-state electrochemiluminescent (ECL) biosensor was fabricated by immobilizing glucose oxidase and surface-unpassivated CdTe quantum dots (QDs) on a screen-printed carbon electrode (SPCE). The surface morphology of the biosensor was characterized with scanning electron microscopy and atomic force microscopy. With dissolved O(2) as an endogenous coreactant, QDs/SPCE showed strong ECL emission in pH 9.0 HCl-Tris buffer solution with low ECL peak potential at -0.89 V. The ECL intensity was twice that with hydrogen peroxide as coreactant at the same concentration. This phenomenon meant the ECL decreased upon consumption of dissolved O(2) and thus could be applied to the construction of oxidase-based ECL biosensors. With glucose oxidase as a model enzyme, the biosensor showed rapid response to glucose with a linear range of 0.8 to 100 μM and a detection limit of 0.3 μM. Further detection of glucose contained in human serum samples showed acceptable sensitivity and selectivity. This work provided a promising application of QDs in ECL-based disposable biosensors.