Modelling of the impedimetric responses of an aflatoxin B1 immunosensor prepared on an electrosynthetic polyaniline platform

Aflatoxins are a group of mycotoxins that have deleterious effects on humans and are produced during fungal infection of plants or plant products. An electrochemical immunosensor for the determination of aflatoxin B₁ (AFB₁) was developed with AFB₁antibody (AFB₁-Ab) immobilized on Pt electrodes modified with polyaniline (PANi) and polystyrene sulphonic acid (PSSA). Impedimetric analysis shows that the electron transfer resistances of the Pt/PANi–PSSA electrode, the Pt/PANi–PSSA/AFB₁-Ab immunosensor and Pt/PANi–PSSA/AFB₁-Ab incubated in bovine serum albumin (BSA) were 0.458, 720 and 1,066 kΩ, respectively. These results indicate that electrochemical impedance spectroscopy (EIS) is a suitable method for monitoring the change in electron transfer resistance associated with the immobilization of the antibody. Modelling of EIS data gave equivalent circuits which showed that the electron transfer resistance increased from 0.458 kΩ for the Pt/PANi–PSSA electrode to 1,066 kΩ for the Pt/PANi–PSSA/AFB₁-Ab immunosensor, indicating that immobilization of the antibody and incubation in BSA introduced an electron transfer barrier. The AFB₁ immunosensor had a detection limit of 0.1 mg/L and a sensitivity of 869.6 kΩ L/mg.     

Comparison of dansylated aminopropyl controlled pore glass solvated by molecular and ionic liquids

We compare how (i) four ionic liquids (ILs) (1-butyl-3-methylimidazolium tetrafluoroborate ([C4mim][BF4]), 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([C4mim][Tf2N]), 1-butyl-1-methylpyrrolidinium bis(trifluoromethylsulfonyl)imide ([C4mpy][Tf2N]), and trihexyltetradecylphosphonium bis(trifluoromethylsulfonyl)imide ([P(C6)3C14][Tf2N])) and (ii) two conventional molecular liquids (methanol and 1-octanol) solvate/wet luminescent organic moieties that are covalently attached to the surface of silica controlled pore glass (CPG). A series of aminopropyl CPG particles that have been covalently tagged with the solvatochromic fluorescent probe group dansyl were used in this study. The results demonstrate that ILs solvate/wet the silica surface differently in comparison to molecular liquids (MLs). Specifically, when comparing ILs and MLs that appear to solvate the free probe, dansylpropylsulfonamide (DPSA), equally in solution, we find that ILs do not solvate/wet the silica surfaces as well as the corresponding MLs. The cation component in these ILs is the significant factor in how the ILs solvate/wet silica surfaces. Solvation/wetting of surface-bound species at a silica surface depends on the cation size. Chlorosilane end-capping of the surface silanol and amine residues attenuates the cation's affects.     

Completely aqueous procedure for the growth of polymer brushes on polymeric substrates

The growth of polymer brushes on polymer substrates is often challenging because of substrate incompatibility with the organic solvents used for initiator attachment. This letter reports the use of layer-by-layer adsorption of macroinitiators and subsequent aqueous ATRP from these immobilized initiators to prepare polymer brushes on polymeric substrates. Polyethersulfone (PES) films and porous membranes were modified with polyelectrolyte multilayer films, and a previously developed polycationic initiator, poly(2-(trimethylammonium iodide)ethyl methacrylate-co-2-(2-bromoisobutyryloxy)ethyl acrylate), was then electrostatically adsorbed onto these polyelectrolyte films. The immobilized macroinitiator is very efficient in initiating the growth of polymer brushes on PES, as demonstrated by aqueous syntheses of poly(2-hydroxyethyl methacrylate) (PHEMA) and poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) films. PHEMA (250 nm thick) and PDMAEMA (40 nm thick) brushes were grown in 2 h from surfaces modified with polycationic initiators. Moreover, this procedure is effective for growing brushes in the pores of PES membranes.     

NMR Study of the Hydrolysis and Oligomerization of Alkyltrichlorosilanes in Silanizing Solutions Used to Prepare Alkylsiloxane Self-Assembled Monolayers

²⁹Si NMR spectroscopy is not suited to following a number of fast occurring processes involving silicon centres due to long accumulation times resulting from low detector sensitivity factors and poor natural abundance. By observing subtle changes in signals due to the methylene protons adjacent to the silicon centre, the hydrolysis of alkyltrichlorosilanes in tetrahydrofuran and acetone-d₆ solutions, and subsequent polycondensation reactions, were studied using ¹H and ¹³C NMR spectroscopy, to gain an understanding of the processes that lead to formation of oligomers in silanizing solutions. The hydroxysilanes formed by hydrolysis of alkyltrichlorosilanes were characterized by ¹H and ¹³C NMR studies at low temperatures. Formation of oligomeric and polymeric species from these hydroxysilanes at higher temperatures was monitored by ¹H NMR studies. The data for oligomer formation were fitted to the kinetic model of an acid-catalyzed stepwise condensation. The implications of these results for the problem of oligomer formation competing with self-assembly processes in formation of alkylsiloxane monolayers are discussed.     

PNA/dsDNA complexes: site specific binding and dsDNA biosensor applications

The ability of peptide nucleic acids (PNA) to form specific higher-order (i.e., three- and four-stranded) complexes with DNA makes it an ideal structural probe for designing strand-specific dsDNA biosensors. Higher-order complexes are formed between a dye-labeled charge-neutral PNA probe and complementary dsDNA. Addition of a light-harvesting cationic conjugated polymer (CCP) yields supramolecular structures held together by electrostatic forces that incorporate the CCP and the dye-labeled PNA/DNA complexes. Optimization of optical properties allows for excitation of the CCP and subsequent fluorescence resonance energy transfer (FRET) to the PNA-bound dye. In the case of noncomplementary dsDNA, complexation between the probe and target does not occur, and dye emission is weak. The binding between PNA and noncomplementary and complementary dsDNA was examined by several methods. Gel electrophoresis confirms specificity of binding and the formation of higher-order complexes. Nano-electrospray mass spectrometry gives insight into the stoichiometric composition, including PNA/DNA, PNA(2)/DNA, PNA/DNA(2), and PNA(2)/DNA(2) complexes. Finally, structural characteristics and binding-site specificity were examined using ion mobility mass spectrometry in conjunction with molecular dynamics. These results give possible conformations for each of the higher-order complexes formed and show exclusive binding of PNA to the complementary stretch of DNA for all PNA/DNA complexes. Overall, the capability and specificity of binding indicates that the CCP/PNA assay is a feasible detection method for dsDNA and eliminates the need for thermal denaturing steps typically required for DNA hybridization probe assays.     

Homogeneous silver-coated nanoparticle substrates for enhanced fluorescence detection

A simple method has been developed for the deposition of uniform silver-coated nanoparticles on glass substrates, with a homogeneous distribution shown by scanning electron microscopy (SEM). UV-visible spectroscopy and energy-dispersive X-ray analysis (EDX) have been used to characterize both the optical density and elemental content of the deposited nanoparticles. The fluorescence enhancement was investigated using a monolayer of FITC-conjugated human serum albumin (FITC-HSA) and tested using laser scanning microscopy at 488 nm excitation wavelength. The enhancement factor was calculated from individual spectra recorded with a Fluorolog-Tau-3 spectrofluorometer. We identified the nanoparticle growth regime which led to fluorescence enhancement. Such enhancement is detectable when Au core-Ag shell nanoparticles increased their size to 47 nm, in agreement with theoretical estimates. The origin of this enhancement for appropriate size nanoparticles is attributed to the effect of an increased excitation rate from the local field enhanced by the interaction of incident light with the nanoparticles and/or higher quantum yield from an increase of the intrinsic decay rate of the fluorophore. We thus demonstrated that the Au core-Ag shell nanostructures on glass surfaces are promising substrates for fluorescence enhancement with outstanding macroscopic homogeneity. This important feature will pave the way for the application of our substrates in biotechnology and life sciences such as imaging and sensing of biomolecules in proteomics.     

Palladium(0)-catalyzed synthesis of chiral ene-allenes using alkenyl trifluoroborates

Enantioenriched allenes serve as chiral transfer reagents, making them attractive synthetic targets. Herein, the synthesis of enantioenriched allenes utilizing a Pd(0)-catalyzed cross-coupling reaction of propargylic carbonates and phosphates with alkenyl trifluoroborates is reported. Di-, tri-, and tetrasubstituted allenes were synthesized in moderate to high optical yields. Several racemic allenes possessing various functional groups were also synthesized.     

Paramagnetic cobalt(II) as an NMR probe of dendrimer structure: mobility and cooperativity of dendritic arms

Cobalt(II) has been utilized as an external paramagnetic (1)H NMR probe for the study of the structure of dendrimers that possess specifically located metal recognition sites. The hyperfine-shifted (1)H NMR signals of the Co(II) complexes of several 2,6-diamidopyridine-containing dendrimers have been fully assigned by means of 1D and 2D NMR techniques, including NOE difference, EXSY, COSY, and TOCSY. Temperature-dependent T(1) values of the hyperfine-shifted signals were used to conclude that the Co(II)-dendrimer complexes are in the "liquidlike" regime, indicative of a shell-like structure instead of a "dense-core" structure. The presence of sizable cavities within the dendrimers was observed including a loosely packed conformation for the 2,6-diamidopyridino moiety to bind to potential guest molecules. Cooperativity among the dendritic arms in metal binding is also observed, whereby two dendritic arms bind to the metal center at the same time. In the case of dendrimers with the metal binding site located near the surface of the molecule, such binding cooperativity is still observed despite the large degree of freedom of the metal-binding moiety. Cooperativity among the dendritic arms can thus be considered an intrinsic property, which has to be taken into consideration in future design of functional dendrimers for the purpose of specific recognition and catalysis. The hydrodynamic radii of these dendrimers have been determined by means of nuclear Overhouser effect at low temperature. The study offers a method for the study of the dynamics of dendrimers in solution under different conditions and upon ligand binding and recognition. The study also provides a tool for monitoring systematic variation of the metal binding site in different dendrimer frameworks for specific applications, such as catalysis and molecular recognition.     

Ionic liquid-induced unprecedented size enhancement of aggregates within aqueous sodium dodecylbenzene sulfonate

Physicochemical properties of aqueous micellar solutions may change in the presence of ionic liquids (ILs). Micelles help to increase the aqueous solubility of ILs. The average size of the micellar aggregates within aqueous sodium dodecylbenzene sulfonate (SDBS) is observed by dynamic light scattering (DLS) and transmission electron microscopy (TEM) to increase in a sudden and drastic fashion as the IL 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF(6)]) is added. Similar addition of [bmim][PF(6)] to aqueous sodium dodecyl sulfate (SDS) results in only a slow gradual increase in average aggregate size. While addition of the IL [bmim][BF(4)] also gives rise to sudden aggregate size enhancement within aqueous SDBS, the IL 1-ethyl-3-methylimidazolium tetrafluoroborate ([emim][BF(4)]), and inorganic salts NaPF(6) and NaBF(4), only gradually increase the assembly size upon their addition. Bulk dynamic viscosity, microviscosity, dipolarity (indicated by the fluorescent reporter pyrene), zeta potential, and electrical conductance measurements were taken to gain insight into this unusual size enhancement. It is proposed that bmim(+) cations of the IL undergo Coulombic attractive interactions with anionic headgroups at the micellar surface at all [bmim][PF(6)] concentrations in aqueous SDS; in aqueous SDBS, beyond a critical IL concentration, bmim(+) becomes involved in cation-π interaction with the phenyl moiety of SDBS within micellar aggregates with the butyl group aligned along the alkyl chain of the surfactant. This relocation of bmim(+) results in an unprecedented size increase in micellar aggregates. Aromaticity of the IL cation alongside the presence of sufficiently aliphatic (butyl or longer) alkyl chains on the IL appear to be essential for this dramatic critical expansion in self-assembly dimensions within aqueous SDBS.