Real-time detection of lymphocytes binding on an antibody chip using SPR imaging

We demonstrate the use of SPR imaging for the detection of site-specific binding of either B or T lymphocyte populations on an electrochemically-grafted antibody array.     

Label-free detection of DNA mutations by SPR: application to the early detection of inherited breast cancer

A screening analysis of DNA hybridization and the presence of DNA mutations using an surface plasmon resonance (SPR) biosensor is shown. The influence of lateral and vertical spacers, as well as several hybridization conditions, was studied to optimize the differentiation between fully complementary and mismatched DNA strands. Our results demonstrated that SPR biosensors were able to detect mismatch sequences related to inherited breast cancer, with high specificity and sensitivity. Using PCR synthetic sequences as targets, mutant sequences were clearly discriminated from fully complementary ones, and detection limits below 50 nM were achieved.     

Surface characterization and efficiency of a matrix-free and flat carboxylated gold sensor chip for surface plasmon resonance (SPR)

We report the preparation and characterization of a matrix-free carboxylated surface plasmon resonance (SPR) sensor chip with high sensing efficiency by functionalizing a bare gold thin film with a self-assembled monolayer of 16-mercaptohexadecanoic acid (SAM–MHDA chip). The self assembled monolayer surface coverage of the gold layer was carefully evaluated and the SAM was characterized by infrared reflection absorption spectroscopy, X-ray photoemission spectroscopy, atomic force microscopy, X-ray reflectivity-diffraction, and SPR experiments with bovine serum albumin. We compared the SPR signal obtained on this chip made of a dense monolayer of carboxylic acid groups with commercially available carboxylated sensor chips built on the same gold substrate, a matrix-free C1 chip, and a CM5 chip with a ~100 nm dextran hydrogel matrix (GE Healthcare). Two well-studied interaction types were tested, the binding of a biotinylated antibody (immunoglobulin G) to streptavidin and an antigen–antibody interaction. For both interactions, the well characterized densely functionalized SAM–MHDA chip gave a high signal-to-noise ratio and showed a gain in the availability of immobilized ligands for their partners injected in buffer flow. It thus compared favourably with commercially available sensor chips.     

Phased fiber growth in a peptide conjugate: aggregation and disaggregation studies

A glycine-rich, short pentapeptide conjugate 6, derived from the highly conserved copper-binding octarepeat region of the prion protein, exhibits a tendency to self-aggregate in a time-dependent fashion. Aging of 6 afforded an insight into the phased growth of spherical prefibrillar structures to fibers of long persistence length, as observed by a combination of microscopic techniques. Interestingly, growth of these fibers was inhibited by colchicine, a known inhibitor of microtubule polymerization in a concentration dependent fashion. This study offers an intriguing insight into the occurrence of prefibrillar intermediates on the path to the formation of full length peptide fibers. It is also envisaged that constructs such as 6 may also serve as simple models to study chemical intervention of protein aggregation.     

SPR imaging biosensor for the 20S proteasome: sensor development and application to measurement of proteasomes in human blood plasma

The 20S proteasome is a multicatalytic enzyme complex responsible for intracellular protein degradation in mammalian cells. Its antigen level or enzymatic activity in blood plasma are potentially useful markers for various malignant and nonmalignant diseases. We have developed a method for highly selective determination of the 20S proteasome using a Surface Plasmon Resonance Imaging (SPRI) technique. It is based on the highly selective interaction between the proteasome’s catalytic β5 subunit and immobilized inhibitors (the synthetic peptide PSI and epoxomicin). Inhibitor concentration and pH were optimized. Analytical responses, linear ranges, accuracy, precision and interferences were investigated. Biosensors based on either PSI and epoxomicin were found to be suitable for quantitative determination of the proteasome, with a precision of ±10% for each, and recoveries of 102% and 113%, respectively, and with little interference by albumin, trypsin, chymotrypsin, cathepsin B and papain. The proteasome also was determined in plasma of healthy subjects and of patients suffering from acute leukemia. Both biosensors gave comparable results (2860 ng·mL-1 on average for control, and 42300 ng·mL-1 on average for leukemia patients).

化学发光检测在微流控芯片中的应用综述

综述了近年来化学发光检测在微流控芯片中的应用.指出微流控芯片(又称为"芯片实验室"或者"微型全分析系统")因具有小型化、集成化和自动化等特点而在近20年来日益受到关注,而化学发光检测具有仪器结构简单、背景噪音低、操作和维护成本低等优点,非常适合用作微流控芯片的检测手段.     

Dynamic interaction between oppositely charged vesicles: aggregation, lipid mixing, and disaggregation

We investigated dynamic interactions between oppositely charged small unilamellar vesicles using positively charged vesicles containing 1,2-dioleoyl-3-trimethylammonium-propane or 3beta-[N-(N('),N(')-dimethylaminoethane)-carbamoyl] cholesterol and negatively charged vesicles containing L-alpha-phosphatidyl-DL-glycerol. Aggregation, lipid bilayer mixing, contents mixing and contents leakage were systematically examined using optical density measurements, fluorescence resonance energy transfer assays, fluorescence quenching assays, light-scattering analyses, and freeze-fracture transmission electron microscopy. The oppositely charged vesicles aggregated immediately. Lipid mixing was observed, but there was no mixing of the contents. The vesicle aggregates disaggregated spontaneously after several minutes. The surface potential of the disaggregated vesicles was neutralized. From these results, we infer that the lipids in the external monolayers were exchanged between the oppositely charged vesicles while the internal monolayers remained intact. The two types of cationic lipids used exhibited different speeds of disaggregation.     

Slow aggregation and disaggregation of TiO2 nanocrystals in aqueous HCl solutions

The kinetics of slow aggregation and disaggregation of the plate-shaped TiO₂ nanocrystals in aqueous HCl solutions at room temperature was studied using turbidimetry and small-angle X-ray scattering. The rate of slow aggregation of the diluted TiO₂ hydrosols with 0.3 ≤ pH ≤ 3 was compared. The sols with pH about 1 were the most stable and the aggregation rate increased at the decrease of pH from 1 to 0.3 and at the increase of pH from 1 to 3. The reversible loose aggregates slowly formed at the low pH, while the more stable aggregates grew at higher pH.     

Rapid detection of a cocaine-binding aptamer using biological nanopores on a chip

This paper describes a methodology for the rapid and highly selective detection of cocaine using a membrane protein channel combined with a DNA aptamer. The DNA aptamer recognizes the cocaine molecule with high selectivity. We successfully detected a low concentration of cocaine (300 ng/mL, the drug test cutoff limit) within 60 s using a biological nanopore embedded in a microchip.     

Electrochemical sensor for estriol hormone detection in biological and environmental samples

A stable conducting film for sensing using reduced graphene oxide (RGO), gold nanoparticles (GNPs), and potato starch (PS) is proposed. The characterization of the nanomaterials was obtained by ultraviolet and visible spectroscopy, dynamic light scattering, zeta potential, Fourier transform infrared spectroscopy, atomic force microscopy, and cyclic voltammetry. The voltammetric behavior of the RGO-GNPs-PS/GCE electrodes was studied in the presence of estriol and the results showed a high anodic peak current at 0.64 V. Under optimal conditions, an analytical curve was obtained, in which the anodic peak estriol was linear in the range from 1.5 to 22 μmol L^?1, with a detection limit of 0.48 μmol L^?1. The modified electrodes were applied for determination of estriol in environmental and biological samples. The proposed electrode was used for estriol determination in water and urine samples, which presented a recovery range from 92.1 to 106%, showing that RGO-GNPs-PS/GCE is a viable alternative for the detection of estriol and can be attractive for several electrochemical applications.     

Simplification and evaluation of a gold-deposited SPR optical fiber sensor.

The structure of the sensing element of a gold-deposited optical fiber sensor was simplified and quantitative analyses of various alcohols with the sensor showed improvement of the performance. The sensor uses surface plasmon resonance (SPR) at the interface of a sample solution and a thin (10 - 70 nm) gold film deposited on half of the exposed core of the optical fiber. The sensor with a film thickness of 45 nm can detect a small change of 5.6 x 10(-5) refractive index (RI) units in the refractivity. The response time is less than 0.5 min and the relative standard deviation for measurements is less than or equal to 1%. A straight line with a correlation coefficient of 0.9995 was obtained below 10%, v/v in the calibration curve for methanol solutions of benzyl alcohol. The minimum of the response curve due to the maximum excitation of SPR in the refractivity range from 1.33 to 1.44 RI units shifts to a lower refractivity as the film becomes thicker. The response curves of the sensors were calculated from SPR theoretical equations while considering of the distribution in the thickness of the deposited gold films. The improvement in the performance of the sensor is discussed.     

Physical adsorption of N,N'-diphenyl-p-phenylenediamine onto carbon particles: application to the detection of sulfide

The derivatisation of carbon powder by physical adsorption of N,N'-diphenyl-p-phenylenediamine (DPPD) onto the surface of graphite particles (1-2 microm diameter) results in usefully functionalised carbon. The derivatised carbon powders have been studied using cyclic voltammetry by (i) abrasive immobilisation of the powder onto a basal plane pyrolytic graphite electrode (bppg) and (ii) by the manufacture of carbon-epoxy electrodes containing the derivatised carbon. The electrochemical response of the DPPD modified carbon has been examined in the presence of sulfide and possible reactions identified for the behaviour of each electrode substrate. The analytical application of the carbon-epoxy electrode for the sensing of sulfide is reported.     

A colorimetric sensor array for detection and discrimination of biothiols based on aggregation of gold nanoparticles

Developments of sensitive, rapid, and cheap systems for identification of a wide range of biomolecules have been recognized as a critical need in the biology field. Here, we introduce a simple colorimetric sensor array for detection of biological thiols, based on aggregation of three types of surface engineered gold nanoparticles (AuNPs). The low-molecular-weight biological thiols show high affinity to the surface of AuNPs; this causes replacement of AuNPs’ shells with thiol containing target molecules leading to the aggregation of the AuNPs through intermolecular electrostatic interaction or hydrogen-bonding. As a result of the predetermined aggregation, color and UV–vis spectra of AuNPs are changed. We employed the digital mapping approach to analyze the spectral variations with statistical and chemometric methods, including hierarchical cluster analysis (HCA) and principal component analysis (PCA). The proposed array could successfully differentiate biological molecules (e.g., cysteine, glutathione and glutathione disulfide) from other potential interferences such as amino acids in the concentration range of 10–800 μmol L⁻¹.     

A coumarinylaldoxime as a specific sensor for Cu and its biological application

In this Letter we present a new probe, (E)-7-(diethylamino)-2-oxo-2H-chromene-3-carbaldehyde oxime (JB), which can detect Cu²⁺ ions in HEPES buffer under physiological conditions. Benesi–Hildebrand and Job plots demonstrate that the stoichiometry of the Cu²⁺ complex formed is 2:1. Possible interference with other analytes was examined, and the decrease of the fluorescence of JB at 510 nm when it reacts with Cu²⁺ was shown to be highly selective. This probe accumulates in the plasmalemma of human neuroblastoma SH-SY5Y cells. Molecular dynamics (MD) simulations revealed that JB interacts with the lipid bilayer at the level of the glycerol moieties.     

利用传感器探究“催化剂对化学反应速率的影响”

在毒品检测方面,常规实验室检测方法已日趋成熟. 现存的实验室分析方法已不能满足毒品现场快速侦测需求,亟待发展便携式毒品探测技术及设备. 在一些快速检测手段中,便携式设备的应用已经普及并且被不断深入研究. 传感器作为一种具有快速识别目标物质的装置,在毒品检测中可以对一些可疑物品进行快速甄别. 随着毒品种类的变化,以及毒品类型的不断更新迭代,对传感器的检测要求也随之增加,从而不同种类和功能的传感器也应运而生. 鉴于传感器具备易于操作、成本较低、快速分析等特点,在禁毒领域尤其是现场检测方面,便携式毒品传感器的应用会成为现场快速检测的重要组成部分. 近年来,随着人们研究的需求化导向及研究内容的不断深入,一些新型毒品探测技术、材料及设备被成功开发. 随着实测信息的反馈及设备的不断优化,便携式毒品探测设备将成为打击毒品犯罪的有力手段.     

Voltammetric sensor based on magnetic particles modified composite electrode for determination of triamterene in biological sample

Some diuretic substances are controlled and monitored by the World Anti-Doping Agency as prohibited substances for use by athletes, such as triamterene (TRT). Thus, this work describes a voltammetric method based on graphite-epoxy composite electrode modified by tosyl-functionalized magnetic particles (GECE/MPs-To) for determination of TRT diuretic in urine sample. The TRT presented an oxidation peak at +1.24 V at GECE/MPs-To with irreversible behavior. Controlled potential electrolysis of the TRT at +1.26 V indicated the two electrons are transferred during amine group oxidation and the main product was identified by LC-MS/MS. The anodic peak current is 25 % higher at the modified electrode, suggesting that TRT is adsorbed on the magnetic particles. Using optimized conditions by using multivariate optimization of the parameters inherent of the square wave voltammetry, a calibration curve was constructed with a linear relationship for TRT from 0.500 to 99.8 μmol L^?1. The limits of detection and quantification were 1.47 and 4.91?×?10^?7 mol L^?1, respectively. The proposed method was applied to urine sample and validated by LC-MS/MS technique where the values found and compared between the two techniques showed no significant difference at 95 % confidence.     

New detection modality for label-free quantification of DNA in biological samples via superparamagnetic bead aggregation

Combining DNA and superparamagnetic beads in a rotating magnetic field produces multiparticle aggregates that are visually striking, enabling label-free optical detection and quantification of DNA at levels in the picogram per microliter range. DNA in biological samples can be quantified directly by simple analysis of optical images of microfluidic wells placed on a magnetic stirrer without prior DNA purification. Aggregation results from DNA/bead interactions driven either by the presence of a chaotrope (a nonspecific trigger for aggregation) or by hybridization with oligonucleotides on functionalized beads (sequence-specific). This paper demonstrates quantification of DNA with sensitivity comparable to that of the best currently available fluorometric assays. The robustness and sensitivity of the method enable a wide range of applications, illustrated here by counting eukaryotic cells. Using widely available and inexpensive benchtop hardware, the approach provides a highly accessible low-tech microscale alternative to more expensive DNA detection and cell counting techniques.