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At least 163 records · Page 9Linked to original sources
PubMed1999

Characteristics of acoustic plate modes on rotated Y-cuts of quartz utilized for biosensing applications.

Acoustic plate modes (APM) on various quartz substrates have been investigated in order to determine their usefulness for liquid-sensing applications. The modes have been characterized in terms of their mass sensitivity, mode separation, temperature sensitivity, and reproducibility of the experimental results. Promising characteristics are found for rotated Y-cuts of quartz with the direction of acoustic mode propagation being perpendicular to the X-axis of the quartz crystal. Experiments on the detection of immunochemical reactions are performed using different quartz APM sensors, and the results are compared to similar experiments utilizing APM devices on ZX-LiNbO3.

Animals↗
PubMedDate not supplied

Development of nanoparticle libraries for biosensing.

Magnetic and magnetofluorescent nanoparticles have become important materials for biological applications especially for sensing, separation, and imaging. To achieve target specificity, these nanomaterials are often covalently modified with binding proteins such as antibodies or proteins. Here we report on the creation of nanoparticle libraries that achieve specificity through multivalent modification with small molecules. We explore different synthetic routes to attach small molecules with anhydride, amine, hydroxyl carboxyl, thiol, and epoxy handles. We show that the derived nanomaterials have unique biological functions, possess different behaviors in cell screens, and can be used as substrates for biological screens.

Amines↗
PubMed2002

Surface-bound lipid vesicles encapsulating redox species for amperometric biosensing of pore-forming bacterial toxins.

A new method is described to signal the presence of a staphylococcal toxin (streptolysin, SLO) by a membrane-based biosensor fabricated with bilayer vesicles of phosphatidylcholine lipids and cholesterol on a gold electrode. The controlled release of encapsulated redox species from the surface-bound vesicles is mediated by the pore-formation functionality of SLO, allowing amperometric detection of the targeted toxin. Effects of BSA and Triton-100 on membrane permeability and redox release have been assessed, and a thin layer electrolysis mechanism has been proposed to account for the observed voltammetric behavior. The method is applicable to the detection of functionally similar protein toxins.

Bacterial Proteins↗
PubMed2004

Ultrasensitive electrical biosensing of proteins and DNA: carbon-nanotube derived amplification of the recognition and transduction events.

A new strategy for dramatically amplifying enzyme-linked electrical detection of proteins and DNA using carbon nanotubes (CNTs) for carrying numerous enzyme tracers and accumulating the enzymatically liberated product on CNT-modified transducer is described. Such a CNT-derived double-step amplification pathway (of both the recognition and transduction events) allows the detection of DNA and proteins down to 1.3 and 160 zmol, respectively, in 25-50 muL samples and indicates great promise for PCR-free DNA analysis. The new protocol is illustrated for monitoring sandwich hybridization and antibody-antigen interactions in connection with alkaline-phosphatase tracers. The DNA-linking of CNTs and particles holds promise also for assembling hybrid nanostructures relevant to molecular electronic devices.

Alkaline Phosphatase↗
PubMed2004

Covalently modified silicon and diamond surfaces: resistance to nonspecific protein adsorption and optimization for biosensing.

We report the direct covalent functionalization of silicon and diamond surfaces with short ethylene glycol (EG) oligomers via photochemical reaction of the hydrogen-terminated surfaces with terminal vinyl groups of the oligomers, and the use of these monolayers to control protein binding at surfaces. Photochemical modification of Si(111) and polycrystalline diamond surfaces produces EG monolayers linked via Si-C bond formation (silicon) or C-C bond formation (diamond). X-ray photoelectron spectroscopy was used to characterize the monolayer composition. Measurements using fluorescently labeled proteins show that the EG-functionalized surfaces effectively resist nonspecific adsorption of proteins. Additionally, we demonstrate the use of mixed monolayers on silicon and diamond and apply these surfaces to control specific versus nonspecific binding to optimize a model protein sensing assay.

Adsorption↗
PubMed2005

Selective functionalization of In2O3 nanowire mat devices for biosensing applications.

A strategy to covalently attach biological molecules to the electrochemically active surface of indium oxide nanowire (In2O3 NW) mat devices is presented. A self-assembled monolayer (SAM) of 4-(1,4-dihydroxybenzene)butyl phosphonic acid (HQ-PA) was generated on an indium tin oxide (ITO)-coated glass and In2O3 NWs surface. The chemical steps required for surface derivatization were optimized on an ITO surface prior to modifying the In2O3 NWs. The hydroquinone group contained in the HQ-PA SAM was electrochemically oxidized to quinone (Q-PA) at +330 mV. The monolayer of Q-PA was allowed to react with a thiol-terminated DNA. The DNA was paired to its complementary strand tagged with a fluorescence dye. Attachment of DNA was verified using fluorescence microscopy. A device was subsequently prepared on a SiO2-supported mat of In2O3 NWs by depositing gold electrodes on the mat surface. The reaction strategy optimized on ITO was applied to this In2O3 NW-based device. Arrays of In2O3 NWs on a single substrate were electrochemically activated in a selective manner to Q-PA. Activated In2O3 NWs underwent reaction with HS-DNA and gave a positive fluorescence response after pairing with the dye-DNA. The unactivated In2O3 NWs gave no response, thus demonstrating selective functionalization of an In2O3 NW array. This can be considered a key step for the future fabrication of large-scale, inexpensive, nanoscale biosensors.

Biosensing Techniques↗
PubMed2005

Biosensing using porous silicon double-layer interferometers: reflective interferometric Fourier transform spectroscopy.

A simple, chip-based implementation of a double-beam interferometer that can separate biomolecules based on size and that can compensate for changes in matrix composition is introduced. The interferometric biosensor uses a double-layer of porous Si comprised of a top layer with large pores and a bottom layer with smaller pores. The structure is shown to provide an on-chip reference channel analogous to a double-beam spectrometer, but where the reference and sample compartments are stacked one on top of the other. The reflectivity spectrum of this structure displays a complicated interference pattern whose individual components can be resolved by fitting of the reflectivity data to a simple interference model or by fast Fourier transform (FFT). Shifts of the FFT peaks indicate biomolecule penetration into the different layers. The small molecule, sucrose, penetrates into both porous Si layers, whereas the large protein, bovine serum albumin (BSA), only enters the large pores. BSA can be detected even in a large (100-fold by mass) excess of sucrose from the FFT spectrum. Detection can be accomplished either by computing the weighted difference in the frequencies of two peaks or by computing the ratio of the intensities of two peaks in the FFT spectrum.

Biosensing Techniques↗
PubMed2006

Potentiometric biosensing of proteins with ultrasensitive ion-selective microelectrodes and nanoparticle labels.

We report here for the first time on the use of potentiometry for ultrasensitive nanoparticle-based detection of protein interactions. A silver ion-selective microelectrode is used to detect silver ions oxidatively released from silver enlarged gold nanoparticle labels in a sandwich immunoassay. Since potentiometry is expected to largely maintain its analytical characteristics upon reducing the sample volume, it is anticipated that this approach may form the basis for bioassays with attractive detection limits.

Biosensing Techniques↗
PubMed2006

Electron donor solvent effects provide biosensing with quantum dots.

A palmitate biosensor that uses the emission intensity of a semiconducting nanoparticle to report palmitate concentration is presented. This method uses electron transfer to quench the emission from a ZnS-coated CdSe nanoparticle. The fatty acid binding pocket of intestinal fatty acid binding protein is used to modulate the electron transfer properties of [Ru(L)(NH3)4](PF6)2 (L = 5-maleimido-1,10-phenanthroline) that is covalently attached within this pocket. Once the metal-complex-modified protein is attached to ZnS-coated CdSe nanoparticles, palmitate addition excludes water from around the metal complex and increases the electron transfer from the metal complex to the valence band hole of the nanoparticle excited state. A 1.6-fold change in emission intensity is observed upon adding a saturated amount (500 nM) of sodium palmitate. The dissociation constant was calculated as 5 nM with a 1 nM lower limit of detection. Since palmitate does not alter the global conformation of intestinal fatty acid binding protein, palmitate-mediated changes in pocket solvation are suggested. This represents a new method in biosensor construction with semiconducting nanoparticles. Including previous conformation-dependent biosensors, there are thousands of potential analytes that can be detected with these strategies. Such biosensors will provide fluorescence contrast imaging reagents for small molecule analytes.

Biosensing Techniques↗
PubMed2005

Mesoporous membrane device for asymmetric biosensing.

An array of micron size reactors has been designed and fabricated on a mesoporous membrane to create a platform for asymmetric biochemical sensing. Fabrication of this device required that a technique be developed to integrate the mesoporous alumina membrane with a polymeric layer that maintains the integrity of the membrane surface and permeability. This device was used to control an enzyme reaction at the surface of the membrane through the diffusion of the substrates from the opposite sides of the membrane. Asymmetric reactions promise new modes of sensing, enhanced stability of delicate biomolecular systems, and enhanced sensitivity and speed in sensing.

Aluminum Oxide↗
PubMed2005

Coadsorption of horseradish peroxidase with thionine on TiO2 nanotubes for biosensing.

In this study, we investigate the coadsorption of protein with thionine on TiO(2) nanotubes for biosensor design. The TiO(2) nanotube arrays fabricated by anodic oxidation of titanium substrate possess large surface areas and good uniformity and conformability and are ready for enzyme immobilization. Electrochemical and spectroscopic measurements show that the TiO(2) nanotube arrays provide excellent matrixes for the coadsorption of horseradish peroxidase (HRP) and thionine and that the adsorbed HRP on these TiO(2) nanotube arrays effectively retains its bioactivity. The immobilized thionine can be electrochemically reduced but cannot be reoxidized in the electrode potential range between -0.7 and 0.0 V. The addition of H(2)O(2) leads to the biocatalytic oxidation of the reduced thionine in the presence of HRP, resulting in developing a novel H(2)O(2) sensor with good stability and reproducibility. The fabricated TiO(2) nanotubes offer a stage for further study of immobilization and electrochemistry of proteins. The proposed method opens a way to develop biosensors using nanostructured materials with low electrical conductivity.

Adsorption↗
PubMed2005

Multilayer membranes via layer-by-layer deposition of organic polymer protected Prussian blue nanoparticles and glucose oxidase for glucose biosensing.

Polyelectrolyte multilayers (PEMs) are now widely used for bioanalytical applications. In this work, a bilayer of poly(diallydimethylammonium chloride) (PDDA) and poly(sodium 4-styrenesulfonate) (PSS) is consecutively adsorbed on 3-mercapto-1-propanesulfonic acid modified Au electrode surfaces, forming stable, ultrathin multilayer films. Subsequently, Prussian blue nanoparticles protected by PDDA (denoted as P-PB) and negatively charged glucose oxidase (GOx) are consecutively adsorbed onto the PSS-terminated bilayer. The growth of each of the P-PB/GOx bilayers is followed quantitatively using UV-visible absorption spectroscopy and the electrochemical method. The P-PB nanoparticles can catalyze the electroreduction of hydrogen peroxide formed from enzymatic reaction at lower potential and inhibit the responses of interferents, such as ascorbic acid (AA) and uric acid (UA). Performance of the multilayer films can be tailored by controlling the number of bilayers. Under optimal conditions, a linear range of 0.10 to 11.0 mM and a detection limit of 10 microM were achieved. The glucose biosensor has good stability and reproducibility.

Biosensing Techniques↗
PubMed2006

Biosensing using lipid bilayers suspended on porous silicon.

We demonstrate for the first time the formation of a fluid lipid bilayer membrane on mesoporous silicon substrates for bioapplications. Using fluorescence recovery after photobleaching, the diffusion coefficients for the bilayers supported on oxidized, amino-, and biotin-functionalized mesoporous silicon were determined. The biodetection of a single human umbilical vein endothelial cell was accomplished using confocal microscopy and exploiting Foerster resonance energy transfer effects after the incorporation of RGD covalently linked lipid soluble dyes, with fluorescence donor and acceptor components, within the fluid membrane. A signal response of greater than 100% was achieved via the clustering of RGD peptides binding with areas of high integrin density on the surface of a single cell. These results are a testament to the usefulness of such functional molecular assemblies, based on mobile receptors, mimicking the cell membrane in the development of a new generation of biosensors.

Biosensing Techniques↗
PubMed2006

Designing highly specific biosensing surfaces using aptamer monolayers on gold.

To build highly specific surfaces using aptamer affinity reagents, the effects of linker and coadsorbents were investigated for maximizing target binding and specificity for aptamer-based self-assembled monolayers (SAMs) supported on gold. An aptamer that binds the protein thrombin was utilized as a model system to compare different mixed monolayer systems toward maximizing binding and selectivity to the immobilized aptamer. Important factors used to optimize binding characteristics of thrombin to the aptamer-based monolayer films include changes in design elements of the linker and different coadsorbent thiols. Binding events measured by surface plasmon resonance (SPR) and ellipsometry showed that the binding performance of the aptamer SAMs depends principally on the linker and to a lesser extent on the coadsorbent. SAMs formed with HS-(CH2)6-OP(O)2O-(CH2CH2O)6-TTTTT-aptamer exhibited a 4-fold increase in binding capacity versus SAMs made using HS-(CH2)6-TTTTT-aptamer. Furthermore, SAMs made using HS-(CH2)6-OP(O)2O-(CH2CH2O)6-TTTTT-aptamer showed nearly complete specificity for thrombin versus bovine serum albumin (BSA, less than 2% bound), while a SAM incorporating a random DNA fragment (HS-(CH2)6-OP(O)2O-(CH2CH2O)6-TTTTT-RANDOM) showed little binding of thrombin. Irrespective of the aptamer-linker system, use of HS-(CH2)11(OCH2CH2)3OH, referred to as EG(3), as a coadsorbent enhanced binding of thrombin by approximately 2.5-fold compared to that of HS-(CH2)6-OH (mercaptohexanol, MCH).

Aptamers, Nucleotide↗
PubMed2006

Zirconia nanoparticles enhanced grafted collagen tri-helix scaffold for unmediated biosensing of hydrogen peroxide.

A novel, biocompatible, thermally steady, and nontoxic zirconia enhanced grafted collagen tri-helix scaffold was prepared on a graphite electrode. This scaffold provided a microenvironment for loading biomolecules and helped to retain their natural structure. UV-vis spectroscopy and scanning electron microscopy were used to characterize the scaffold and the structure of immobilized biomolecules. Using horseradish peroxidase (HRP) as an example, this scaffold accelerated its electron transfer and led to its direct electrochemical behavior with a good thermal stability up to 80 degrees C. The surface electron-transfer rate constant of the immobilized HRP was (5.55 +/- 0.43) s(-)(1) in 0.1 M pH 7.0 PBS at 18 degrees C. The immobilized HRP showed an electrocatalytic activity to the reduction of hydrogen peroxide (H(2)O(2)) without aid of an electron mediator. The linear response range of the biosensor for H(2)O(2) was from 1.0 to 73.0 microM with a correlation coefficient of 0.999 (n = 14), a limit of detection down to 0.25 microM and an apparent Michaelis-Menten constant of (0.28 +/- 0.02) mM. The biosensor exhibited high sensitivity, acceptable stability, and reproducibility. The ZrO(2) grafted collagen provided an excellent matrix for protein immobilization and biosensor preparation.

Biosensing Techniques↗
PubMedDate not supplied

Fluorescence-based biosensing of zinc using carbonic anhydrase.

Measurement of free zinc levels and imaging of zinc fluxes remains technically difficult due to low levels and the presence of interfering cations such as Mg and Ca. We have developed a series of fluorescent zinc indicators based on the superb sensitivity and selectivity of a protein, human apo-carbonic anhydrase II, for Zn(II). These indicators transduce the level of free zinc as changes in intensity, wavelength ratio, lifetime, and/or anisotropy; the latter three approaches permit quantitative imaging of zinc levels in the microscope. A unique attribute of sensors incorporating biological macromolecules as transducers is their capability for modification by site-directed mutagenesis. Thus we have produced variants of carbonic anhydrase with improved affinity for zinc, altered selectivity, and enhanced binding kinetics, all of which are difficult to modify in small molecule indicators.

Animals↗
PubMed2004

Label free colorimetric biosensing using nanoparticles.

In this review article, we discuss a class of biosensors that exploit the change in the colorimetric properties of noble metal nanoparticles in response to biomolecular binding at their surface. Several sensor fabrication techniques as well as sensor configurations are discussed with an emphasis on their strengths and limitations. We conclude by presenting the future prospects and challenges for the successful transition of this technology from the laboratory to a commercial product.

Algorithms↗