Yoko Ishii

Highly sensitive detection of platelet-derived growth factor on a functionalized diamond surface using aptamer sandwich design

Aptamer-based fluorescence detection of platelet-derived growth factor (PDGF) on a functionalized diamond surface was demonstrated. In this work, a sandwich design based on the ability of PDGF to bind with aptamers at its two available binding sites was employed. It was found that this sandwich design approach significantly increases the fluorescence signal intensity, and thereby a very low detection limit of 4 pM was achieved. The effect of the ionic strength of MgCl(2) buffer solution was also investigated, and the most favourable binding for PDGF-BB occurred at a Mg(2+) concentration of 5.5 mM. Since the aptamers bind to the target PDGF with high affinity, fluorescence detection exhibited high selectivity towards different biomolecules. The high reproducibility of detection was confirmed by performing three cycles of measurements over a period of three days.

Effective surface functionalization of nanocrystalline diamond films by direct carboxylation for pdgf detection via aptasensor

An aptasensor was designed on a nanocrystalline diamond (NCD) surface that combined with biological recognition elements, PDGF-binding aptamers, which inherently possess high affinity to PDGF-BB proteins. Functional components such as carboxylic acids (-COOH) and amines (-NH2) were directly introduced onto the NCD surface and used as probing units for immobilization of PDGF-binding aptamers. The surface coverage of different components on the NCD was analyzed by X-ray photoelectron spectroscopy (XPS) measurements, and the effects of various functionalizations on the NCD biosensor performance were investigated via fluorescence observations. The coverages of carboxyl and amine groups achieved were 12 and 23%, respectively, for the directly aminated and carboxylated NCD; however, the lower density of carboxyl groups on the functionalized surface did not deteriorate the performance of the COOH-NCD biosensor. Fluorescence investigations demonstrated comparable performance in sensitivity and selectivity for PDGF protein detection on COOH-NCD and NH2-NCD biosensors. Multiple regeneration tests clearly showed that the COOH-NCD biosensor as well as the NH2-NCD biosensor retained a high performance without exhibiting any noticeable degradation.

Aptamer-based biosensor for sensitive PDGF detection using diamond transistor.

The detection of platelet-derived growth factor (PDGF) via a solution-gate field-effect transistor (SGFET) has been demonstrated for the first time using aptamers immobilized on a diamond surface. Upon introduction of PDGF to the immobilized aptamer, a shift of 31.7 mV in the negative direction is observed at a source-drain current of -50 μA. A shift of 32.3 mV in the positive direction is detected after regeneration by SDS solution, indicating that the static measurement returns to its original value. These SGFETs operate stably within the large potential window of diamond (>3.0 V), and hence the surface channel does not need passivating with a thick insulating layer. Thereof, the immobilized aptamer channels have been exposed directly to the electrolyte solution without a gate insulator. Immobilization is achieved via aptamers covalently bonding to amine sites, thereby increasing the sensitivity of the biosensors. Diamond SGFETs have potential for the detection of PDGF and show durability against biological degradation after repeated usage and regeneration.

Human immunodeficiency virus trans-activator of transcription peptide detection via ribonucleic acid aptamer on aminated diamond biosensor

The potential of ribonucleic acid (RNA) as both informational and ligand binding molecule have opened a scenario in the development of biosensors. An aminated diamond-based RNA aptasensor is presented for human immunodeficiency virus (HIV) trans-activator of transcription (Tat) peptide protein detection that not only gives a labeled or label-free detection method but also provides a reusable platform for a simple, sensitive, and selective detection of proteins. The immobilized procedure was based on the binding interaction between positively charged amine terminated diamond and the RNA aptamer probe molecules with the negatively charged surface carboxylic compound linker molecule such as terephthalic acid.

Aptasensor for oncoprotein platelet-derived growth factor detection on functionalized diamond surface by signal-off optical method

The highly sensitive detection of platelet-derived growth factor (PDGF) has been realized using aptamers immobilized on a diamond surface. The detection mechanism of PDGF is based on the release of an intercalating dye from the aptamers stem structure during deformation when the aptamer captures PDGF. The detection by fluorescence reduction (signal-off method) does not require a target label and is unaffected by nonspecific binding. The high reproducibility of detection was confirmed. Since aptamers bind to their target PDGF with high affinity, the sensor exhibited high selectivity and sensitivity. The quantitation limit was determined to be 1 pM.

Gas permeation of LC films observed by smectic bubble expansion

Gas permeation through liquid crystal (LC) films was examined using hemispherical smectic bubbles. A smectic bubble, when the inside and the outside are filled with different gases, should expand or shrink toward the quasi-equilibrium state, where the influx and efflux caused by osmotic pressure are balanced. Deriving a simple formula that directly converts the quasi-equilibrated bubble radius to the gas permeation, we determined the absolute permeability coefficients of 8 simple gases through the smectic bubble. The permeability was distributed in such a wide range that carbon-dioxide had more than 20 times larger value than nitrogen, the dependence of which on the gas species was mostly dominated by their solubility into the LCs. Dividing the measured permeability by the calculated solubility, we obtained the diffusion constants as well, yet whose magnitude and the dependence on the solute size could not be explained by either conventional continuum theories or microscopic diffusion models. In order to describe the diffusion of small solutes in the liquid solvent composed of large molecules, a new theoretical framework may be necessary.

Anomalous deformation of smectic bubbles under DC electric field

Hemispherical bubbles composed of smectic liquid crystals show an unusual deformation under DC electric field. When the lower voltage than a threshold is applied to the bubble on a substrate, it is elongated in the direction of the electric field and forms a semi-elliptical shape in the equilibrium. With the higher voltage than the threshold, the bubble becomes unstable and sometimes exhibits a periodic oscillation with repeating the elongation and shrinkage. Based on a simple equation of force balance, we analyzed both the static and dynamic deformations under DC field and successfully reproduced the observed behaviour. By the detailed analysis, we also determined the surface tension and the electric conductivity of the smectic bubbles.