Label-free monitoring of immuno-specific interactions of adsorbed multilayer of proteins.

Protein-protein interactions in adsorbed multilayer of an immuno-specific system of proteins that include staphylococcal protein A (SpA), bovine serum albumin (BSA), anti-chicken immunoglobulin Y (ac-IgG), chicken serum IgG (cs-IgG), and rabbit serum IgG (rs-IgG) on polystyrene (PS) were studied using attenuated total reflection-Fourier transform infrared spectroscopy. A systematic analysis allowed a direct qualitative and quantitative determination of protein interactions at each step of specific and nonspecific binding conditions at the molecular level. The study also provided information about (1) the adsorption behavior of the proteins, (2) the role of SpA in enabling correct orientation of the adsorbed IgG and maintaining the stability of the adsorbed SpA/ac-IgG system on the PS surface, (3) the function of BSA as both blocking reagent and promoter of specific and selective binding, and (4) the bioactivity conserved accommodation of SpA molecules on the PS surface.

Site-specific electronic structure of imidazole and imidazolium in aqueous solutions.

The occupied and unoccupied electronic structure of imidazole (CNH) and imidazolium (CNH) in aqueous solutions is studied by X-ray emission spectroscopy (XES) and resonant inelastic soft X-ray scattering (RIXS). Both systems show distinct RIXS fingerprints with strong resonant effects. A comparison with calculated X-ray emission spectra of isolated imidazole and imidazolium suggests only a small influence of hydrogen bonding in the aqueous solution on the electronic structure of imidazole and imidazolium, and allows the attribution of specific spectral features to the non-equivalent nitrogen and carbon atoms in the molecules.

X-ray Emission Spectroscopy of Proteinogenic Amino Acids at All Relevant Absorption Edges.

Nonresonant N K, O K, C K, and S L X-ray emission spectra of the 20 most common proteinogenic amino acids in their solid zwitterionic form are reported. They represent a comprehensive database that can serve as a reliable basis for the X-ray absorption spectroscopy (XES) studies of peptides and proteins. At the most important N and O K edges, clear similarities and differences between the spectra of certain amino acids are observed and associated with the specific chemical structure of these molecules and their functional groups.

Investigation of the Ionic Hydration in Aqueous Salt Solutions by Soft X-ray Emission Spectroscopy.

Understanding the molecular structure of the hydration shells and their impact on the hydrogen bond (HB) network of water in aqueous salt solutions is a fundamentally important and technically relevant question. In the present work, such hydration effects were studied for a series of representative salt solutions (NaCl, KCl, CaCl2, MgCl2, and KBr) by soft X-ray emission spectroscopy (XES) and resonant inelastic soft X-ray scattering (RIXS). The oxygen K-edge XES spectra could be described with three components, attributed to initial state HB configurations in pure water, water molecules that have undergone an ultrafast dissociation initiated by the X-ray excitation, and water molecules in contact with salt ions.

Ion-Solvation-Induced Molecular Reorganization in Liquid Water Probed by Resonant Inelastic Soft X-ray Scattering.

The molecular structure of liquid water is susceptible to changes upon admixture of salts due to ionic solvation, which provides the basis of many chemical and biochemical processes. Here we demonstrate how the local electronic structure of aqueous potassium chloride (KCl) solutions can be studied by resonant inelastic soft X-ray scattering (RIXS) to monitor the effects of the ion solvation on the hydrogen-bond (HB) network of liquid water. Significant changes in the oxygen K-edge emission spectra are observed with increasing KCl concentration.

Surface-confined heterometallic triads on the basis of terpyridyl complexes and design of molecular logic gates.

Surface-confined heterometallic molecular triads (SURHMTs) were fabricated on SiOx-based solid substrates using optically rich and redox-active Fe-, Os-, and Ru-based terpyridyl complexes as metalloligands and Cu(2+) ions as linkers. Optical and electrochemical studies reveal efficient electronic intramolecular communication in these assemblies. The UV-vis spectra of the triads exhibit a superposition of the metal-to-ligand charge-transfer bands of individual complexes, providing a significant enlargement of the optical window, useful for application.

Surface confined heteroleptic copper(II)-polypyridyl complexes for photonuclease activity.

Heteroleptic copper(II)-polypyridyl complexes with extended π-conjugated, aromatic terminal units were immobilized on glass/Si substrates to intercalate DNA and cleave it upon photoexposure. Photonuclease activity is shown to be high, well reproducible and non-destructible towards the assembled complexes.

Modification of polystyrene surface in aqueous solutions.

Herein, we report our analysis of the surface modification of polystyrene (PS) when treated under ambient conditions with a common biological buffer such as phosphate buffered saline (PBS) or aqueous solutions of the ionic constituents of PBS. Attenuated total reflection Fourier transform infrared spectroscopy was used for the analysis because the resultant spectra are very sensitive to minor changes in the chemical and structural properties of PS films. In addition, ultraviolet-visible spectroscopy was applied to characterize the surface modifications of PS.

A simple method of surface functionalisation for immuno-specific immobilisation of proteins.

We present a new and advanced methodology, developed for surface functionalisation of gold and to study immobilisation of an immuno-specific system of proteins. A combination of electrochemical quartz crystal microbalance and Raman spectroscopy techniques allowed a complete understanding of the system starting from surface functionalisation and progressing to the functional structure analysis of immobilised proteins. A simple electrochemical procedure was formulated to prepare sulphonyl chloride terminated gold surfaces that form a strong sulphonamide bond with the receptor protein staphylococcal protein A (SpA).

Efficiency of blocking of non-specific interaction of different proteins by BSA adsorbed on hydrophobic and hydrophilic surfaces.

The efficiency of a pre-absorbed bovine serum albumin (BSA) layer in blocking the non-specific adsorption of different proteins on hydrophobic and hydrophilic surfaces was evaluated qualitatively and quantitatively using infrared reflection spectroscopy supported by spectral simulations. A BSA layer with a surface coverage of 35% of a close-packed monolayer exhibited a blocking efficiency of 90-100% on a hydrophobic and 68-100% on a hydrophilic surface, with respect to the non-specific adsorption of concanavalin A (Con A), immunoglobulin G (IgG), and staphylococcal protein A (SpA). This BSA layer was produced using a solution concentration of 1 mg/mL and 30 min incubation time.

Quantitative and qualitative evaluation of adsorption/desorption of bovine serum albumin on hydrophilic and hydrophobic surfaces.

We studied the adsorption of bovine serum albumin (BSA) from phosphate-buffered saline (pH 7.4) to hydrophilic and hydrophobic surfaces. Attenuated total reflection Fourier transform infrared spectroscopy, supported by spectral simulation, allowed us to determine with high precision the amount of BSA adsorbed (surface coverage) and its structural composition.

The effect of surface composition of titanium films on bacterial adhesion.

We show that bacterial adhesion on titanium (Ti) films could be radically minimized by tailoring the surface chemical stoichiometry of the films. Using a dc magnetron sputtering system, Ti films with various surface compositions, such as oxide and nitride combinations, were prepared by controlling processing parameters such as cathode power, sputtering pressure and base vacuum. The surface topography of the films was observed to be smooth and similar in all the films prepared under different conditions.