Formation of tetra(ethylene oxide) terminated Si-C linked monolayers and their derivatization with glycine: an example of a generic strategy for the immobilization of biomolecules on silicon

Surface modification with oligo(ethylene oxide) functionalized monolayers terminated with reactive headgroups constitutes a powerful strategy to provide specific coupling of biomolecules with simultaneous protection from nonspecific adsorption on surfaces for the preparation of biorecognition interfaces. To date, oligo(ethylene oxide) functionalized monolayer-forming molecules which can be activated for attachment of biomolecules but which can selectively form monolayers onto hydrogen terminated silicon have yet to be developed. Here, self-assembled monolayers (SAMs) containing tetra(ethylene oxide) moieties protected with tert-butyl dimethylsilyl groups were formed by thermal hydrosilylation of alkenes with single-crystal Si(111)-H. The protection group was used to avoid side reactions with the hydride terminated silicon surface. Monolayer formation was carried out using solutions of the alkene in the high-boiling-point solvent 1,3,5-triethylbenzene. The protecting group was removed under very mild acidic conditions to yield a free hydroxyl functionality, a convenient surface moiety for coupling of biological entities via carbamate bond formation. The chemical composition and structure of the monolayers before and after deprotection were characterized by X-ray photoelectron spectroscopy (XPS) and X-ray reflectometry. To demonstrate the utility of this surface for covalent modification, two reagents were compared and contrasted for their ability to activate the surface hydroxyl groups for coupling of free amines, carbonyl diimidazole (CDI), and disuccinimidyl carbonate (DSC). Analysis of XP spectra before and after activation by CDI or DSC, and after subsequent reaction with glycine, provided quantitative information on the extent of activation and overall coupling efficiencies. CDI activated surfaces gave poor coupling yields under various conditions, whereas DSC mediated activation followed by aminolysis at neutral pH was found to be an efficient method for the immobilization of amines on tetra(ethylene oxide) modified surfaces.     

Studies of solid-state ion-selective electrodes prepared from semiconducting organic radical-ion salts

The primary redox reactions for solid-state ion-selective electrodes prepared from electronically semiconducting salts of 7,7,8,8-tetracyanoquinodimethane (tcnq) can be identified by considering the redox properties of their constituent ions or molecules. Three different processes involving the couples, Mⁿ⁺/M⁰, 2tcnq^o/(tcnq^-)₂ and (tcnq^-)₂/2tcnq^2- are possible depending on salt composition. Ionic product values determined by potentiometric and atomic absorption methods are in excellent agreement for several such salts; K_s(K₂tcnq₂)=5.8±1.2·10^-11(pot.), 1.7±1·10^-11 (a.a.s.); K_s(Cdtcnq₂) = 3.0±0.5·10^-9 (pot.), 2.9±0.3·10^-9(a.a.s.); K_s(Pbtcnq₂) = 1.3±0.3·10^-10 (pot.), 0.96±0.2·10^-10(a.a.s.); and indicate that the lower activity limit for electrode response is controlled by the solubility of the sensor material itself. Comparisons of predicted and observed standard electrode potentials provide quantitative support for an ion-exchange mechanism of interference. The behaviour of electrodes prepared from Cu₂tcnq₂ (copper(I)) and Cutcnq₂ (copper(II)) is explained on the basis of an interference mechanism and considerations of solid-state equilibria.     

Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking

CE conditions for monitoring the unsaturated disaccharides of hyaluronic acid (di-HA) and chondroitin sulfate (di-CS) using an alkaline tetraborate buffer, electrokinetic sample injection, and UV absorption detection at 232 nm are reported. Separations were performed in an uncoated fused-silica capillary having reversed polarity and reversed electroosmosis generated with the addition of CTAB to the buffer. The influence of various separation parameters, including the concentration of CTAB, buffer pH, concentration of tetraborate, and applied voltage, on the resolution of the two disaccharides was investigated. Baseline separation was obtained with 25 mM tetraborate at pH 10.0 and having 0.05 mM CTAB. Chloride and phosphate in the sample are beneficial for the stacking of the disaccharides, with di-HA forming a much sharper peak than di-CS. Using samples prepared in 25 mM Tris-HCl (pH 7.5) and electrokinetic injection at the cathode at -10 kV for 40 s, linear relationships between the corrected peak area and the concentration of the disaccharides have been found in the ranges of 1.0-400.0 and 0.1-1.0 microg/mL (0.2-1.0 microg/mL for di-CS), with correlation coefficients being >0.9933 in all cases. The RSDs of detection times and corrected peak areas were between 1.13-1.24 and 1.57-2.13%, respectively. Applied to human serum samples that were prepared by ethanol precipitation and depolymerization of the two polysaccharides with chondroitinase ABC reveals comigration of endogenous compounds with di-HA and a sample-dependent detection time. The di-HA content in the serum sample can be estimated via subtraction of the blank peak that is obtained without enzymatic hydrolysis.     

Enhanced rates of electrolytic styrene epoxidation catalyzed by cross-linked myoglobin-poly(L-lysine) films in bicontinuous microemulsions

Redox proteins attached to surfaces designed for biocatalysis hold promise for future clean synthetic routes. It is advantageous for these biocatalysts to operate in low-toxicity fluids with a high capacity to dissolve reactants. Here we report cross-linked films of myoglobin (Mb) and poly(L-lysine) (PLL) chemically attached to oxidized carbon cloth cathodes that in microemulsions feature the protein in a water-rich film environment with reactant in an oil-rich environment. These cross-linked Mb/PLL films were the most stable in microemuslions and had the largest turnover rates for epoxidation of styrene compared to lightly cross-linked or uncross-linked Mb/poly(styrene sulfonate) films. Up to 40-fold larger turnover rates were found in bicontinuous microemulsions compared to oil-in-water microemulsions and micelles. Enhanced turnover rates are correlated with up to 10-fold faster mass transport of solutes in the oil phases of the bicontinuous fluids.     

Dendritic molecular capsules for hydrophobic compounds

Reichardt's dye, a highly solvatochromic dye, was encapsulated within poly (glycerol succinic acid) ([Gn]-PGLSA-OH) dendrimers to investigate the interior environment of these dendritic macromolecules. The absorption maximum for the encapsulated Reichardt's dye in water was indicative of a relatively high dielectric constant present within the dye/dendrimer complex. (1)H NMR of the encapsulated complex showed the presence of aromatic protons from Reichardt's dye along with the aliphatic protons of the dendrimer. Additionally, there were substantial changes in T(1) and T(2) times of the encapsulated dye when compared with the free dye, and (1)H NOESY spectra for the complex showed a significant number of intermolecular NOE cross-peaks. These data reveal the close through-space proximity of the dye to the dendrimer and the restricted motion of the encapsulated dye. To demonstrate the potential use of these macromolecules as drug delivery vehicles, the poorly water-soluble anticancer drug 10-hydroxycamptothecin (10HCPT) was encapsulated within a carboxylated PGLSA dendrimer ([G4]-PGLSA-COONa). Cytotoxicity assays with human breast cancer cells showed a significant reduction of cell viability, demonstrating that 10HCPT retains activity upon encapsulation.     

Microchemical techniques as applied to radiochemistry

Microchimica Acta - The techniques for investigating the chemistry of radioactive elements usingμg quantities of material are described. These include methods of transfer and carrying out...     

Reversible Michael addition of thiols as a new tool for dynamic combinatorial chemistry

The Michael addition of thiols to enones is reported as a new method for dynamic combinatorial library synthesis.