Surface plasmon resonance

The elimination or minimization of non-specific protein adsorption from serum is critical for the use of surface plasmon resonance (SPR) sensors for in vitro and in vivo analysis of complex biological solutions. The ultimate goals in this application are to minimize non-specific adsorption of protein and to maximize analyte signal. A reduction of the non-specific protein adsorption from serum of

Aim. Comparison of the IgG-binding activity of recombinant Staphylococcal protein A with introduced C-terminal cysteine residue (SPA-Cys) or goat anti-human IgG antibodies (anti-IgG) after their immobiliza-tion on a gold sensor surface of surface plasmon resonance (SPR) spectrometer. Methods. SPA-Cys or anti-IgG were immobilized on a gold sensor surface to form two variants of a bioselective element of the immu-nosensor. SPR spectrometry was used for the detection of IgG-binding activity of the immobilized proteins. Results. The SPR sensor response to the immobilization of anti-IgG was more than two times higher than that to the immobilization of SPA-Cys. However, there is almost the double advantage for SPA-Cys in the number of immobilized molecules. Moreover, the bioselective element of the immunosensor based on SPA-Cys showed a much better capability of binding IgG than the bioselective element based on anti-IgG. Conclusions. Comparison of immobilization of SPA-Cys or anti-IgG on the sensor surface of SPR spectrometer, and the interactions of immobilized proteins with human IgG demonstrated obvious advantages of SPA-Cys. K e y w o r d s: antibodies, recombinant Staphylococcal protein A, protein immobilization, immunosensor, surface plasmon resonance.

Interaction of fibrin with endothelial cells through their receptor VE-cadherin has been implicated in modulation of angiogenesis and inflammation. Previous studies identified the VE-cadherin-binding site in the fibrin βN-domains formed by the NH2-terminal regions of fibrin β chains, and revealed that the recombinant dimeric (β15-66)2 fragment mimicking these domains preserves the VE-cadherin-binding properties of fibrin. To test if the other fibrin(ogen) regions/domains are involved in this interaction and localize the complementary fibrin-binding site in VE-cadherin, we prepared several recombinant fragments containing individual extracellular domains of VE-cadherin or combinations thereof, as well as several fragments corresponding to various fibrin(ogen) regions, and tested the interactions between them by ELISA and surface plasmon resonance. The experiments revealed that the βN-domains are the only fibrin(ogen) regions involved in the interaction with VE-cadherin. They also loc...

While most microbes play an important role in nature, certain potentially harmful microbes can contaminate food and water, and cause a plethora of infectious diseases in both animals and humans. Conventional methods for detecting microbial contamination have primarily ...

Fully characterizing the interactions involving biomolecules requires information on the assembly state, affinity, kinetics, and thermodynamics associated with complex formation. The analytical technologies often used to measure biomolecular interactions include analytical ultracentrifugation (AUC), isothermal titration calorimetry (ITC), and surface plasmon resonance (SPR). In order to evaluate the capabilities of core facilities to implement these technologies, the Association of Biomolecular Resource Facilities (ABRF) Molecular Interactions Research Group (MIRG) developed a standardized model system and distributed it to a panel of AUC, ITC, and SPR operators. The model system was composed of a well-characterized enzyme-inhibitor pair, namely bovine carbonic anhydrase II (CA II) and 4-carboxybenzenesulfonamide (CBS). Study participants were asked to measure one or more of the following: (1) the molecular mass, homogeneity, and assembly state of CA II by AUC; (2) the affinity and ...

Bulk characteristics and deposition of gold nanoparticles on mica modified by (PAH) was studied byUV–visible spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM),dynamic light scattering (DLS) and atomic force microscopy (AFM). The size of gold nanoparticles (AuNPs)was 15 nm as determined by TEM and AFM. The electrophoretic mobilities and electrokinetic chargeof particles were quantitatively evaluated for a broad range of pH and ionic strength. Using Henry’smodel, it was calculated that the zeta potential of particles varied between − 50 mV and − 80 mV forpH 2 and pH 11, respectively (at 10−2M of NaCl). Measurements of nanoparticle deposition kineticswere performed under diffusion-controlled transport conditions using AFM and SEM imaging of particlemonolayers. The influence of the bulk suspension concentration was systematically studied. Additionally,the maximum coverage of particle monolayers, which monotonically increased with ionic strength, wasdetermined by SEM. The obtained data were in agreement with theoretical predictions derived from therandom sequential adsorption (RSA) model. It was also confirmed that by varying the bulk suspensionconcentration and ionic strength one can prepare homogeneous gold particle monolayers of controlledcoverage.

Gold nanoparticles have been attracting attention due to their extensive application in chemistry, physics, material science, electronics, catalysis and bionanotechnology. Synthesis of gold nanoparticles often involves toxic and expensive physical-chemistry methods. Preparation of gold nanoparticles by arc discharge in water is proposed for the first time. Fabrication of gold nanostructures in deionized water has been successfully established. The evidence of gold particles’ light absorbance reveals a unique surface plasmon resonance for Au nanoparticles suspended in deionized water. Gold nanostructures uniformly dispersed in water, their UV–Vis absorption and crystalline size are shown. Our experimental results demonstrate that fabrication of gold nanoparticles by arc discharge in water is an alternative, cheap, effective and environmentally friendly method.

The selection of nanoparticles for achieving efficient contrast for biological and cell imaging applications, as well as for photothermal therapeutic applications, is based on the optical properties of the nanoparticles. We use Mie theory and discrete dipole approximation method to calculate absorption and scattering efficiencies and optical resonance wavelengths for three commonly used classes of nanoparticles: gold nanospheres, silica-gold nanoshells, and gold nanorods. The calculated spectra clearly reflect the well-known dependence of nanoparticle optical properties viz. the resonance wavelength, the extinction cross-section, and the ratio of scattering to absorption, on the nanoparticle dimensions. A systematic quantitative study of the various trends is presented. By increasing the size of gold nanospheres from 20 to 80 nm, the magnitude of extinction as well as the relative contribution of scattering to the extinction rapidly increases. Gold nanospheres in the size range comm...