Voltammetric detection of avidin and biotin by biotin labeled with cysteine

An avidin-biotin assay was developed from a voltammetric procedure using biotin labeled with cysteine. Mercury(II) as a marker was used to detect avidin and biotin, because the oxidation wave of mercury decreases when the cysteine part of labeled biotin(LB) complexes with mercury(II).The formation of the mercury(II)-cysteine complex is suppressed when the LB binds to the biotin site of avidin. Accordingly, the concentration of avidin can be estimated from the increasing mercury peak current. Detection of biotin is also carried out by a competitive reaction of biotin and the LB to the binding site on avidin, where the addition of biotin decreases the peak current of mercury. Limits of detection for avidin and biotin were in the 10^–9 mol/L range. The length of the spacer between the cysteine and biotin was investigated. It was observed that the strength of binding increased with increasing length of spacer. Size considerations rules out steric influences, so it is suggested that the binding constant depends on hydrophobic interactions in the binding site.     

Procedure for controlling size of silver particles deposited photochemically on quartz

The results of studying optical properties of silver films deposited photochemically on a quartz surface are presented. The films consist of isolated silver particles of 5–50 nm in size and of agglomerates of several micrometers in size and contain up to 7 wt % of Ag₂O. Treatment of the films with dimethylformamide solution of [Pden₂](BPh₄)₂ results in a decrease in the average particle size due to dissolution of small particles (5–15 nm) and destruction of greater agglomerates.     

A radial-partition fluoroimmunoassay for the detection of Hepatitis B surface antigen with the streptavidin/biotin complex

A sandwich technique based on the very affinity between avidin and biotin is used. Antibodies are covalently bound to glass-fibre on which the antigen is captured. The discs are then treated in sequence with monoclonal antibodies of high selectivity, species-specific anti-IgG (biotin-linked), streptavidin and β-galactosidase, (biotin-linked). The discs are then incubated with the substrate, 4-methylumbelliferyl-β-d-galactopyranoside and the fluorescence is measured in an automatic immunoassay system.     

Quenching of TiO2 photo catalysis by silver nanoparticles

The plasmon resonance of metal nanostructures affects neighboring semiconductors, quenching or enhancing optical transitions depending on various parameters. These plasmonic properties are currently investigated with respect to topics such as photovoltaics and optical detection and could also have important consequences for photocatalysis. Here the effect of silver nanoparticles of a size up to 30 nm and at maximum 0.50 monolayers on the photocatalytic oxidation of ethylene on TiO₂ is studied. Since the plasmon resonance energy of silver nanoparticles is comparable with the TiO₂ band gap, dipole-dipole interaction converts excitons into heat at the silver nanoparticle. This indicates that plasmonic interaction with TiO₂ semiconductor catalysts can reduce the photo catalytic activity considerably.     

Highly enhanced electrochemiluminescence based on synergetic catalysis effect of enzyme and Pd nanoparticles for ultrasensitive immunoassay

An immunosensor based on the electrochemiluminescence (ECL) of luminol was proposed by coupling enzymatic reaction to in situ generate coreactant with Pd nanoparticles as catalyst for the ECL reaction, which was successfully applied for the ultrasensitive detection of α-1-fetoprotein with a low detection limit of 33 fg mL(-1).     

Effective catalysis and antibacterial activity of silver and gold nanoparticles biosynthesized by Phlogacanthus turgidus

In this work, an environmentally friendly and cost-effective synthetic method of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) was successfully performed using aqueous extract of Phlogacanthus turgidus (PT) leaves. The biosynthesis of nanoparticles was optimized for reaction conditions including concentration of metallic ions, temperature, and time using the measurement of UV–Vis spectroscopy. The nanoparticles were well characterized by analytic techniques such as Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), high-resolution transmission electron microscope (HR-TEM), and selected area electron diffraction (SAED). The morphological data showed that PT-AgNPs possessed the spherical shape with the size distribution ranging from 5 to 15 nm with a mean size of 10 nm while PT-AuNPs existed in the multiple shape with the size distribution ranging from 5 to 20 nm with a mean size of 12 nm. The antibacterial behavior showed that PT-AgNPs possessed high bioactivity against four bacterial strains including Bacillus subtilis, Staphylococcus aureus, Salmonella typhi, and Escherichia coli. Moreover, the catalytic activity of the biogenic nanoparticles was investigated for catalytic reduction of 2-nitrophenol, 3-nitrophenol, and rhodamine B. The kinetic data showed that the nanoparticles were excellent catalysts with potential applications for environmental treatment.

Graphical abstract

     

Asymmetric cyclizations via a sequential Michael addition/Conia-ene reaction by combining multifunctional quaternary phosphonium salt and silver catalysis

A one-pot asymmetric Michael addition/Conia-ene reaction sequence, catalyzed by combination of a dipeptide-derived multifunctional quaternary phosphonium salt and Ag₂CO₃ has been developed, which provides a series of synthetically important chiral methylenecyclopentane derivatives in moderate to excellent yields (up to 97%) and enantioselectivities (up to 93%).     

Study of the affinity of thermographic additives for silver by time-of-flight static secondary ion mass spectrometry and surface-enhanced Raman spectroscopy on silver nanoparticles

Detection of the interactions between low molecular weight organic compounds and metals in the form of sols on a nanoscale is analytically challenging. This study aims to provide experimental evidence using a combination of UV-Vis absorption spectrometry, surface-enhanced Raman spectrometry (SERS), and static secondary ion mass spectrometry (S-SIMS). The field of application is thermography where silver images are formed via heat-catalyzed reactions. Several organic compounds called tone modifiers and stabilizers are used in thermographic materials for the optimization of the image quality. With exploitation of the strengths of each of the above-mentioned methods, an affinity ranking of several tone modifiers and a stabilizer was established on the basis of competitive adsorption experiments using different model systems. Specifically, silver sols, SERS probes, and sputter-coated silver substrates were exposed to systems with one or two additives. The UV-Vis results provided insight on the aggregation of silver nanoparticles in a hydrosol, which was necessary for the interpretation of the SERS data. Both SERS and S-SIMS measurements led to a similar ranking of the relative affinity of the additives in two components, which was largely consistent with empirical knowledge derived from macroscopic behavior.     

Separation method based on affinity reaction between magnetic and nonmagnetic particles for the analysis of particles and biomolecules

A separation method is reported for particle and biochemical analysis based on affinity interactions between particle surfaces under magnetic field. In this method, magnetic particles with immunoglobulin G (IgG) or streptavidin on the surface are flowed through a separation channel to form a deposition matrix for selectively capturing nonmagnetic analytes with protein A or biotin on the surface due to specific antigen (Ag)--antibody (Ab) interactions. This separation method was demonstrated using model reactions of IgG--protein A and streptavidin-biotin on particle surface. The features of this new separation method are (1) the deposited Ag-Ab complex can be examined and further analyzed under the microscope, (2) a kinetic study of complex binding is possible, and (3) the predeposited matrix can be formed selectively and changed easily. The detection limits were about 10(-11) g. The running time was less than 10 min. The selectivities of studied particles were 94% higher than those of label-controlled particles. This method extends the applications of analytical magnetapheresis to nonmagnetic particles. Preliminary study shows that this separation method has a great potential to provide a simple, fast, and selective analysis for particles, blood cells, and immunoassay related applications.     

Capture and detection of a quencher labeled oligonucleotide by poly(phenylene ethynylene) particles

Fluorescence quenching of poly(phenylene ethynylene) (PPE) particles by a Cy-5 labeled oligonucleotide is 2 orders of magnitude more sensitive than direct excitation of the Cy-5 fluorophore.     

A new chemically amplified electrochemical system for the detection of biological affinity reactions: direct and competitive biotin assay

Quantitation of biological affinity reactions by a newly developed chemically amplified electrochemical detection method was demonstrated with the biotin-avidin binding pair. In the method, ruthenium tris(2,2'-bipyridine)(Ru-bipy) was used as an electrochemical signal-generating tag. Its oxidation current on an indium tin oxide (ITO) electrode was amplified with a sacrificial electron donor, oxalate. Because oxalate itself produced negligible current on the electrode, the signal-to-background ratio was greatly enhanced in comparison with other chemical amplification systems. Although the Ru-bipy/oxalate redox couple has been employed previously in electrochemiluminescent and photoelectrochemical detection, its use in a catalytic amperometric detection of biological binding assays has not been reported. To implement the method in the detection of biotin-avidin recognition, avidin was immobilized on an ITO electrode, and was reacted with biotin in solution. Immobilization of avidin by passive adsorption was found to be relatively stable under the condition of the affinity reaction. In the direct assay, biotin labelled with Ru-bipy was recognized by avidin and accumulated on the electrode surface, which was then detected electrochemically in the presence of oxalate. A linear relationship between electrochemical current and biotin concentration was obtained in the range of 1-300 ng mL(-1). In the competitive assay, a mixed solution of unlabelled biotin (the analyte) of various concentrations and 100 ng mL(-1) labelled biotin was reacted with avidin on the surface. As the concentration of the unlabelled biotin increased, less labelled biotin bound to avidin, leading to a reduction in the electro-catalytical response of Ru-bipy. A detection limit of 1 ng mL(-1) biotin was obtained in the competitive assay, which is close to the sensitivity of some enzyme-labelled amperometric assays.     

Potential barrier field-flow fractionation for the separation and characterization of colloidal particles

Summary The reversibility of adsorption of colloidal particles on the channel wall in Sedimentation Field-Flow Fractionation (SFFF), which is based on the variation of the ionic strength of the carrier solution, suggests a new method, for the separation and characterization of colloidal materials. This new method has been called Potential Barrier Field Flow Fractionation (PBFFF).     

Directing stem cell fate by controlling the affinity and density of ligand-receptor interactions at the biomaterials interface

Sticky situation: the differentiation of mesenchymal stem cells can be influenced by the affinity and density of an immobilized ligand for the integrin receptors. Cells adherent to monolayers that present the high-affinity, cyclic-RGD peptide (left) show increased expression of osteogenic markers, while cells on monolayers presenting the lower-affinity, linear-RGD peptide (right) express early markers of myogenesis at a high density and neurogenesis at a low density of the ligand.     

Enhancement of the emission intensity of fluorophore-labeled avidin by biotin and biotin derivatives. Evaluation of different fluorophores for improved sensitivity

Fluorescein, Texas Red, Cascade Blue, 7-amino-methylcoumarin-3-acetic acid, and Lucifer Yellow were evaluated as fluorescent labels for homogeneous fluorophore-linked binding assays. Conjugates of avidin with these fluorophores exhibited an enhancement in fluorescence emission in the presence of biotin or biotin derivatives. This property was used in the development of assays for biotin. The biotin-induced fluorescence enhancement of each labeled avidin were compared. Fluorescein led to the most sensitive calibration (dose-response) curve for biotin with a detection limit of 8 x 10(-10)M.     

Immobilization of laccase by Cu2+ chelate affinity interaction on surface modified magnetic silica particles and its use for the removal of pentachlorophenol

Magnetic Cu²⁺-chelated silica particles using polyacrylamide as a metal-chelating ligand was developed and used for the immobilization of laccase by coordination.The effect of pH and temperature on the enzymatic property of immobilized laccase and its catalytic capacity for pentachlorophenol（PCP） degradation were evaluated systemically.Compared with free laccase,the immobilized laccase showed improved acid adaptabihty and thermal stability.The immobilized laccase prepared in this work exhibited a good catalytic capacity for PCP removal from aqueous solutions.     

Quenched electrochemiluminescence of Ag nanoparticles functionalized g-C3N4 by ferrocene for highly sensitive immunosensing

In this paper, a novel sandwich electrochemiluminescence (ECL) immunosensor was constructed by ferrocene for quenching Ag nanoparticles functionalized g-C₃N₄ (Ag@g-C₃N₄) emission. The prepared Ag@g-C₃N₄ had strong and stable ECL signals compared to pure g-C₃N₄ and primary antibody (Ab₁) can be immobilized on Ag@g-C₃N₄ by adsorption of Ag nanoparticles. Ferrocene carboxylic acid (Fc-COOH) labeled secondary antibody was immobilized on Au doped mesoporous Al₂O₃ nanorods (Au@Al₂O₃–Fc-COOH@Ab₂) as labels through adsorption ability of Au toward proteins. After a sandwich-type immunoreaction, a remarkable decrease of ECL signal was observed due to the ECL quenching of Ag@g-C₃N₄ by Au@Al₂O₃–Fc-COOH@Ab₂. As a result, the change of ECL intensity has a direct relationship with the logarithm of CEA concentrations in the range of 1 pg mL⁻¹–100 ng mL⁻¹ with a detection limit of 0.35 pg mL⁻¹ (S/N = 3). Additionally, the proposed immunosensor shows high specificity, good reproducibility, and long-term stability.