Layer-by-layer assembly of multilayer films composed of streptavidin and biotinylated antibody by real-time biomolecular interaction analysis

Layer-by-layer assembly of multilayer films of streptavidin and biotinylated antibody was completed on the streptavidin coated surface. Real-time biomolecular interaction analysis (BIA) based on surface plasmon resonance technique was used to monitor the multilayer assembly in solution continuously. The results indicate that the uniform multilayer film can be fabricated successfully based on the strong interaction between streptavidin and biotin. The mean surface mass concentration of each adsorption layer is 1. 32 ng/mm(2) for biotinylated antibody, 2. 93 ng/mm(2) for streptavidin, according to the correlation of SPR response with surface concentration.

Layer-by-layer assembly of multilayer films consisting of silicotungstate and a cationic redox polymer on 4-aminobenzoic acid modified glassy carbon electrode and their electrocatalytic effects

A novel 4-aminobenzoic acid (4-ABA) monolayer film is formed on glassy carbon electrode (GCE) by amino cation radical method. Silicotungstic heteropolyanion (SiW12O404-, denoted as SiW12)-containing multilayer films have been fabricated on the 4-ABA modified GCE surface by alternate deposition with a quaternized poly(4-vinylpyridine) partially complexed with [Os(bpy)(2)Cl](2+/+) (denoted as QPVP-Os). Cyclic voltammetry (CV), X-ray photoelectron spectroscopy (XPS) and X-ray reflectivity (XR) have been used to characterise the as-prepared multilayer films. It is proved that the multilayer films are uniform and stable. The average thickness for a bilayer of QPVP-Os/SiW12 in the multilayer film is 30.2 Angstrom. The electrocatalytic activities of the multilayer films have been investigated on the reduction of three substrates of important analytical interests, HNO2, BrO3- and H2O2. Especially, the influence of layer number of the multilayer films on the electrocatalytic reduction of HNO2 has been investigated in detail. (C) 2000 Elsevier Science B.V. All rights reserved.

Assembly of alternating polycation and DNA multilayer films by electrostatic layer-by-layer adsorption

The assembly of alternating DNA and positively charged poly(dimethyldiallylammonium chloride) (PDDA) multilayer films by electrostatic layer-by-layer adsorption has been studied. The real-time surface plasmon resonance (BIAcore) technique was used to characterize and monitor the formation of multilayer films in solution in real time continuously. Electrochemical impedance spectroscopy (EIS) and UV-vis absorbance measurements were also used to study the film assembly, and linear film growth was observed. All the results indicate that the uniform multilayer can be obtained on the poly(ethylenimine)- (PEI-) coated substrate surface. The kinetics of the adsorption of DNA on PDDA surface was also studied by the real-time BIAcore technique; the observed rate constant was calculated using a Langmuir model (k(obs) = (1.28 +/- 0.08) x 10(-2) s(-1).

Multilayer assembly of Mn-substituted Keggin-type [ZnW11O39Mn(H2O)](8-) on chemically modified glassy carbon electrode and its electrocatalytic properties

Through layer-by-layer assembly, undecatungstozincates monosubstituted by transition metals Mn, ZnW11 Mn (H2O) O-39(8-) was successfully immobilized on a glassy carbon electrode surface grafted covalently by 4-aminobenzoic acid. The electrochemical behavior of these polyoxometalates was investigated. Cyclic voltammetry proves the uniform growth of the film. They exhibit some special electrochemical properties in the films, different from those in homogeneous aqueous solution. The effect of pH on the redox behavior of ZnW11Mn(H2O)O-39(8-) in the film was discussed in detail. The multilayer film electrodes have an excellent electrocatalytic response to the reduction of H2O2 and BrO3-, and to the oxidation of ascorbic acid.

Gold-coated iron (Fe@Au) nanoparticles: Synthesis, characterization, and magnetic field-induced self-assembly

A unique reverse micelle method has been developed to prepare gold-coated iron (Fe@Au) nanoparticles. XRD, UV/vis, TEM, and magnetic measurements are utilized to characterize the nanocomposites. XRD only gives FCC patterns of gold for the obtained nanoparticles. The absorption band of the Fe@Au colloid shifts to a longer wavelength and broadens relative to that of the pure gold colloid. TEM results show that the average size of Fe@Au nanoparticles is about 10 nm, These nanoparticles are self-assembled into chains on micron scale under a 0.5 T magnetic field. Magnetic measurements show that the particles are superparamagnetic with a blocking temperature (T-B) of 42 K, At 300 K (above T-B), no coercivity (Hc) and remanence (M-r) is observed in the magnetization curve, while at 2K (below T-B) He and M, are observed to be 728 Oe and 4.12 emu/g, respectively, (C) 2001 Academic Press.

Electrostatic layer-by-layer assembly of polycation and DNA multilayer films by real-time surface plasmon resonance technique

The assembly of alternating DNA and positively charged poly(dimethyldiallylammonium chloride) (PDDA) multilayer films by electrostatic layer-by-layer adsorption has been studied. Real time surface plasmon resonance (BIAcore) technique was used to characterize and monitor the formation of multilayer films in solution in real time continuously. The results indicate that the uniform multilayer can be obtained on the poly(ethylenimine) (PEI) coated substrate surface. The kinetics of the adsorption of DNA on PDDA surface was also studied by real-time BIAcore technique, and the observed rate constant was calculated using a Langmuir model (k(obs) = (1.28 +/- 0.08) x 10(-2) s(-1)).

The electrochemical behavior of polypyrrole from self-assembly polymerization

The electrochemical behavior and the charge transport of polypyrrole film prepared by self-assembly polymerization have been investigated. Ir is found that the microstructure of the film influences the electrochemical behavior of polypyrrole, and that the p-toluenesulfate (Tos(-)) ion plays avery important role in this system.

Self-assembly of the pre-formed inclusion complexes between cyclodextrins and alkanethiols on gold electrodes

A new kind of self-assembled monolayer (SAM) formed in aqueous solution through the pre-formed inclusion complexes (abbreviated CD . C-n) between alpha-, beta-cyclodextrins (CDs) and alkanethiols (CH3(CH2)(n-1)SH, n = 10, 14 and 18) was prepared successfully on gold electrodes. High-resolution H-1 NMR was used to confirm the formation of CD . C-n. X-ray photoelectron spectroscopy, cyclic voltammetry and chronoamperometry were used to characterize the resulting SAMs (denoted as M-CD . Cn). It was found that M-CD . Cn were more stable against repeated potential cycling in 0.5 M H2SO4 than SAMs of CH3(CH2)(n-1)SH (denoted as M-Cn), with a relative sequence of Mbeta-CD . Cn > Malpha-CD . Cn > M-Cn. In addition, an order of blocking the electron transfer between gold electrodes and redox couples (both Fe(CN)(6)(3-) and Ru(NH3)(6)(3+)) in solution, M-CD . C10 > M-CD . C14 > M-CD . C18, was observed. A plausible explanation is provided to elucidate some of the observations. (C) 1997 Elsevier Science S.A.

Spontaneous assembly of the inclusion complexes of viologen-attached alkanethiols and alpha-, beta-cyclodestrin on gold electrode

Electroactive self-assembled monolayers (SAMs) with well-defined electrochemical responses were prepared by spontaneous assembly of the inclusion complexes (CD/C8VComegaSH) of viologen-attached alkanethiols (C8VComegaSH) and alpha- and beta-cyclodextrin (CD). They were characterized by X-ray photoelectron spectroscopy and cyclic voltammetry. The results demonstrate that the chemisorption process of CD/C8VComegaSH on gold substrate occurs through S-Au bonds, and that the redox sites in SAMs of CD/C8VComegaSH are in a much more uniform environment than those in SAMs of C8VComegaSH.

Interfacial characteristics of the self-assembly system of poly-L-lysine 3-mercaptopropionic acid gold electrode

The interfacial characteristics of poly-L-lysine (PL) attached on self-assembled monolayers (SAMs) of 3-mercaptopropionic acid (MPA) were studied by an electrochemical method. The results indicated that PL\MPA layer inhibited partly the diffusion process of redox species in solution, and the electrode surface behaved like a microelectrode array. Its permeation effect was also strongly affected by Mg2+. The more Mg2+ ions were added into the electrolyte solution, the greater the difficulty with which the electron transfer of potassium ferricyanide took place. The three different conformations of PL on the electrode surface had different influences on the electron transfer processes of ferricyanide. PL in random coil state hindered most strongly the electron transfer behavior of ferricyanide,while the alpha-helical PL had nearly no effect and the effect of the beta-sheet state PL was intermediate of these. (C) 1997 Elsevier Science S.A.

Study on Ordered Assembly of SnO2 Nonoparticles-Arachidic Acid Composite LB Films

The hydrosol of SnO2 nanoparticles (NP) have been prepared by colloid chemistry method. The composite LB monolayer and multilayer of SnO2 NP-AA have been obtained by LB technique at the gas-liquid interface of the hydrosol subphase. The structures of the monolayer and multilayer were characterized by IR, UV-Vis, small angle X-ray diffraction spectroscopy and TEM technique, The results indicate that the coverage of SnO2 NP at the composite monolayer's surface is high and the sites of SnO2 NP are similar. The multilayer has good periodic structure.

Studies on the electron transfer centers of amino oxidase immobilized on self-assembly monolayer using electrochemical methods

The direct electron transfer of amino oxidase on electrode surface based on self-assembly technique occurs at 505 mW(vs. Ag/AgCl), indicating that copper atoms are the electron transfer centers and catalytic centers of amino oxidase.

Assembly of the subcellular parts of Bryopsis hypnoides Lamouroux into new protoplasts

The chloroplasts, mitochondria, and protoplasm devoid of mature chloroplasts (PMC) of Bryopsis hypnoides Lamouroux were isolated by low-speed and sucrose density centrifugation. The PMC aggregated in artificial seawater, and then protoplasts without mature chloroplasts (PtMCs) were formed. Transmission electron microscopy and cytochemical studies indicated that there were mitochondria, nuclei, vesicles, and other small cell organelles in the PtMCs. Scanning electron microscopy showed that there were holes on the surface of 1-h PtMCs and then fewer holes on the surface of 24-h PtMCs, suggesting that a healing process occurred. The plasma membrane was formed over the surface of the PtMCs. However, the cell wall was not regenerated, and the newly formed PtMCs were ruptured and died in 3 days. Light intensity during alga maintenance before use influenced significantly (one-way ANOVA, P < 0.0001) on the number of PtMCs formed; the highest number of PtMCs was formed at 20A mu mol/(m(2) s). When isolated chloroplasts were transferred into seawater, there were only two or three chloroplasts aggregated together. However, isolated mitochondria and the mixed six layers of cell organelles (separated by sucrose density centrifugation) could not aggregate in the artificial seawater. This indicates that the conjunction of cell organelles is important for their aggregation.

In vitro assembly of R-phycoerythrin from marine red alga Polysiphonia urceolata

Scanning tunneling microscope was used to investigate the in vitro assembly of R-phycoerythrin (R-PE) from the marine red alga Polysiphonia urceolata. The results showed that R-PE molecules assembled together by disc-to-disc while absorbing on HOPG surface, which just looked like the rods in the phycobilisomes. When the water-soluble R-PE was dissolved in 2% ethanol/water spreading solution, they could form monolayer film at the air/water interface. Similar disc-to-disc array of R-PE was constituted in the two-dimensional Langmuir-Blodgett film by the external force. It could be concluded that, apart from the key role of time linker polypeptides, the in vivo assembly of phycobiliproteins into phycobilisomes is also dependent on the endogenous properties of phycobiliprotein themselves.

Assembly of the protoplasm of Codium fragile (Bryopsidales, Chlorophyta) into new protoplasts

The cell organelles of the coenocytic alga Codium fragile (Sur.) Hariot aggregated rapidly and protoplasts were formed when its protoplasm was extruded out in seawater. Continuous observation showed that there were long and gelatinous threads connecting the cell organelles. The threads contracted, and thus the cell organelles aggregated into protoplasmic masses. The enzyme digestion experiments and Coomassie Brilliant Blue and Anthrone stainings showed that the long and gelatinous threads involved in the formation of the protoplasts might include protein and saccharides as structure components. Nile Red staining indicated that the protoplast primary envelope was non-lipid at first, and then lipid materials integrated into its surface gradually. The fluorescent brightener staining indicated that the cell wall did not regenerate in the newly formed protoplasts and they all disintegrated within 72 h after formation. Transmission electron microscopy of the cell wall of wild C. fragile showed electron-dense material embedded in the whole cell wall at regular intervals. The experiments indicated that C. fragile would be a suitable model alga for studying the formation of protoplasts.