Hydrogel network entrapping cholesterol oxidase and octadecylsilica for optical biosensing in hydrophobic organic or aqueous micelle solvents

Two optical cholesterol biosensors have been fabricated by immobilizing cholesterol oxidase (ChOx) and octadecylsilica (ODS) particles in hydrogel network matrixes of copolymer of poly(vinyl alcohol) (PVA)/hydroxyethyl carboxymethyl cellulose (HECMC), and sol-gel, respectively. In conjunction with an optical oxygen transducer, the immobilized ChOx in the sol-gel/ODS matrix was assembled as an optical cholesterol biosensor to continuously detect free cholesterol in aqueous micelle solution, while the immobilized ChOx in the PVA/HECMC/ODS matrix was constructed as an organic-phase optical cholesterol biosensor for the continuous analysis of free cholesterol in hydrophobic organic solvent. The compositions and properties of the immobilization matrixes, the effects of solvents and the analytical features were studied in detail. Both biosensors showed stable and reliable responses toward free cholesterol. For the aqueous micelle cholesterol biosensor, the analytical working range was from 0.05 to 8.0 mM cholesterol, the response time was 7-12 min, the operation life was more than 35 assays, and the shelf life was approximately 4 months. For the organic-phase cholesterol biosensor, the analytical working range was from 0.07 to 18.0 mM cholesterol, the response time was 4-8 min, the operation life was more than 120 assays, and the shelf life was longer than 5 months. The organic-phase cholesterol biosensor has been successfully applied to determine the free cholesterol content in commercial butter samples.     

辣根过氧化物酶在多壁碳纳米管——壳聚糖膜中的电化学和生物传感性质

采用壳聚糖包裹的多壁碳纳米管膜固定辣根过氧化物酶(HaP)于玻碳电极表面,实现了HRP的直接电化学并以此酶膜制备了NO生物传感器.在磷酸缓冲溶液中固定在电极表面的HRP氧化还原式电位为-0.354 V(us.SCE),直接电子转移速率常数为4.24±1.02 s-1.研究结果表明,固定在电极表面的HRP能保持其对一氧化氮还原的生物电催化活性,该传感器在NO浓度为1.0×10-4~1.4×10-3 mol L-1范围内存在线性响应,响应时间小于11 s,NO的检出限为7×10-4 mol L-1.多壁碳纳米管特殊的电学性质和壳聚糖良好的生物相容性性使得构筑的HRP生物传感器呈现了良好的应用前景,尤其适用于痕量NO的检测.     

Preparation of porous titania sol-gel matrix for immobilization of horseradish peroxidase by a vapor deposition method

A new and facile vapor deposition method has been developed for the preparation of sol-gel matrix. This method was used to form a titania sol-gel thin film and to immobilize horseradish peroxidase (HRP) on a glassy carbon electrode surface for the production of an amperometric hydrogen peroxide biosensor. This process prevented the cracking of conventional sol-gel-derived glasses. The morphologies of both titania sol-gel and the enzyme membranes were characterized using scanning electron microscopy and proved to be chemically clean, porous, and homogeneous and to have a very narrow particle size distribution. The sol-gel-derived titania-modified electrode retained the enzyme bioactivity and provided for long-term stability of the enzyme in storage. In the presence of catechol as a mediator, the sensor exhibited a rapid electrocatalytic response (less than 5 s), a linear calibration range from 0.08 to 0.56 mM with a detection limit of 1.5 microM and a high sensitivity (61.5 microA mM(-1)) for monitoring of H2O2. Effects of pH and operating potential were also explored for optimum analytical performance by using the amperometric method. The apparent Michaelis-Menten constant of the encapsulated HRP was 1.89 +/- 0.21 mM.     

Electrochemically mediated reduction of horseradish peroxidase by 1,1'-ferrocenedimethanol in organic solvents

Cyclic voltammetry is an efficient means of analyzing the catalytic reduction of H2O2 at immobilized horseradish peroxidase (HRP)-Eastman AQ 55 electrodes in the presence of 1,1'-ferrocenedimethanol as a one-electron reversible cosubstrate. This system was employed to study the kinetics of the reduction of compound II of HRP in a number of organic solvents. An electrocatalytic response was detected in methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, acetone, 2-butanone, 1,2-propanediol, acetonitrile, ethyl acetate, and ethylene glycol. Unusual bell-shaped variations of the peak or plateau catalytic current with the substrate concentration were observed in all solvents tested. The results obtained in methanol, acetonitrile, and 1-propanol were analyzed using the model developed by Saveant (Limoges, B.; Saveant, J.-M.; Yazidi, D. J. Am. Chem. Soc. 2003, 125, 9192-9203). The values of k3Gamma0 and K3,M, where k3 = k3,1k3,2/(k3,-1 + k3,2), Gamma0 is the surface concentration of active enzyme, and K3,M = (k3,-1 + k3,2)/k3,1, were determined. The values of k3Gamma0 for the mediated reduction of compound II of HRP in methanol, 1-propanol, and acetonitrile (in the presence of 5% aqueous buffer) were not affected by the solvent dielectric constant but decreased with solvent hydrophobicity. The value of K3,M obtained in methanol was similar to that obtained for [Os(bpy)2pyCl]2+ in aqueous buffer.     

A micro-system based on glass-nanoporous silicon for optical sensing of organic solvent vapor

We present a recent experimental study on the application of nanoporous silicon (np-Si) to an optical vapor sensor. We fabricated the micro-system based on a glass-nanoporous silicon layer on a p(+)-type silicon wafer. To check the selectivity and sensitivity of the np-Si layer to organic vapors, we prepared three types of np-Si layer samples--a single layer, distributed Bragg reflector (DBR) layer, and microcavity layer--and investigated its reflectance spectra upon exposure to different concentrations of various organic vapors. When the np-Si layer samples were exposed to the organic vapors, a red-shift occurred in the reflectance spectrum, and we determined that this red-shift can be attributed to the changes in the refractive index induced by the capillary condensation of the organic vapor within the pores of the np-Si layer. The np-Si layer samples showed excellent sensing ability to different types and concentrations of organic vapors. After removing the organic vapors, the reflectance spectrum immediately returned to its original state.     

Electrical and optical properties of an alcohol soluble aminoalkyl-substituted cationic conjugated polymer

The optical and electrical properties of a novel alcohol soluble aminoalkyl-substituted cationic conjugated polymer, poly[9,9′-bis(6″-(N,N,N-trimethylammonium)-hexyl)fluorene-co-alt-2,5-dimethoxy-1,4-phenylene dibromide] (PFPB), has been studied using absorption, photoluminescence, current–voltage–luminescence (J–V–L), and noise characterization techniques. The absorption and photoluminescence studies show that PFPB is blue-emitting, and its long alkyl side chains and two methoxy side chains introduce steric hindrance in the structure, which can minimize interchain interactions. The photoluminescence quantum efficiency of PFPB in methanol was found to be 69.6%. Polymer light emitting diodes (PLEDs) were fabricated using PFPB as an electron injection layer and device physics has been studied. The obtained device results indicate that PFPB can be used as an electron transport layer (ETL) to improve the PLED performance. The noise characteristics on the PLED devices show that the interface between the cathode and the emissive layer is improved through the introduction of a PFPB layer. Results in this report indicate that the poly(fluorene-co-phenylene)-based water/alcohol soluble cationic conjugated polymer avoids the intermixing between the electroluminance layer and adjacent ETL layer which is a common and serious problem in multilayer PLED fabrication by solution casting methods. This makes water solubility materials attractive for applications in PLEDs and other organic devices.     

A genetically engineered fusion protein with horseradish peroxidase as a marker enzyme for use in competitive immunoassays

Horseradish peroxidase is one of the most widely used marker enzymes in immunoassays. Several disadvantages are encountered upon chemical conjugation of peroxidase with antibodies or antigens, as are low reproducibility and undefined stoichiometry. We here describe for the first time the production of a recombinant fusion of a protein analyte with horseradish peroxidase in Escherichia coli, employing refolding of inclusion bodies and reconstitution with heme. The genetic fusion approach enables preparation of conjugates with 1:1 stoichiometry and defined structure. As a protein analyte, the human heart fatty acid binding protein (H-FABP) was chosen, which is a new and sensitive marker for acute myocardial infarction. The recombinant conjugate was fully active [650 U/mg with 2,2-azino-bis(3-ethyl-thiazoline-6-sulfonate) as substrate] and obtained in a yield of 12 mg/L of E. coli culture, which is better than that for recombinant peroxidase alone. The competitive immunoassay that was developed with the recombinant conjugate requires fewer incubation steps than the traditional sandwich ELISA format. It permitted the detection of H-FABP directly in plasma in the range of 10-1500 ng/mL which is the relevant range for clinical decision-making.     

Multichannel biosensing platform of surface-immobilized gold nanospheres for linear and nonlinear optical imaging

What we believe to be a new label-free multichannel biosensing platform is proposed. It is composed of surface-immobilized gold nanospheres (SIGNs) above a gold surface with a nanogap supported by a merocyanine self-assembled monolayer. The circular SIGN spots with a diameter of 120 microm were arrayed for multichannel biosensing on a glass slide. Two kinds of sensing methods were examined: One is a reflectivity measurement of a blue ray and the other is a second-harmonic generation measurement. It was found that the SIGN system can be used as a promising platform for multichannel biosensing in both sensing methods.     

High-power and high-gain organic dye-doped polymer optical fiber amplifiers: novel techniques for preparation and spectral investigation

Rhodamine B (RB), Rhodamine 6G (R6G), Oxazine 4 perchlorate (O4PC), and 4-Dicyanomethylene-2-methyl-6-(p-dimethylaminostyryl-4H-pyran)-doped graded-index (GI) polymer optical fiber amplifiers (POFA's) were successfully prepared by novel techniques to prevent degradation of organic dyes and to enhance solubility of the dyes in methyl methacrylate and in poly(methyl methacrylate) (PMMA). High-power (620 W) and high-gain (33-dB, 2070 times) amplification was achieved for RB-doped GI POFA's based on the spectral investigation of dyes in PMMA bulk. It should be noted that amplification in the low-loss region of polymer optical fibers was achieved in the R6G- and O4PC-doped GI POFA.     

In situ encapsulation of horseradish peroxidase in electrospun porous silica fibers for potential biosensor applications

Nanoporous silica nanofibers have been employed as a matrix to encapsulate horseradish peroxide enzymes via a simple electrospinning method. A viscous solution of prehydrolyzed tetramethyl orthosilicate, beta-d-glucose, poly(vinyl alcohol), and enzymes were employed as spinning solution to generate porous fibers in the form of nonwoven mats. The silica fiber mats thus produced have a high surface area because of the small diameter (100 to 200 nm) of the fibers as well as the extreme porosity (2 to 4 nm) of individual fibers caused by the glucose template present in them. The high surface area, mechanical flexibility, thermal stability, reusability, and freedom of encapsulating various enzymes make porous silica nanofibers excellent biosensors.     

固定化脂肪酶聚合物膜的改性及脂肪酶膜反应器的应用

综述了在聚合物膜上固定化脂肪酶的方法及脂肪酶膜反应器的应用,重点讨论了纤维素膜,聚丙烯腈膜和砜膜表面的化学改性途径及其与脂肪酶的偶联方法。     

聚乙烯醇/壳聚糖/明胶水凝胶的合成及其对苯酚的吸附研究

以聚乙烯醇(PVA)、壳聚糖和明胶为原料、戊二醛为交联剂,在酸性溶液中通过共混交联反应合成了聚乙烯醇/壳聚糖/明胶水凝胶,单因素法讨论了影响水凝胶性能的因素如:反应物质量比、交联剂用量、反应温度、反应时间等聚乙烯醇/壳聚糖/明胶水凝胶,对不同浓度苯酚溶液的吸附研究.研究结果表明聚乙烯醇/壳聚糖/明胶水凝胶对苯酚有较好的吸附性能.     

Second-order hyperpolarizability and nonlinear optical properties of novel organic compound-doped poly (O-methoxyaniline) polymer film

Journal of Materials Science: Materials in Electronics - The structural characterization of a new organic compound (3, 5-bis (4-chlorophenyl)-4,5-dihydro-1H-Pyrazole) was analyzed experimentally by...     

天然有机高分子及其改性产品在污水处理中的应用

天然有机高分子及其改性产品以其来源广、价廉、残留毒性小的独特优势在污水处理中广泛用作絮凝剂、重金属吸附剂。文章着重介绍了近年来研究较多的淀粉类、甲壳素 /壳聚糖类在污水处理中的应用 ,同时还介绍了其他天然有机高分子及其改性产品在该行业中的应用。     

Enzymatic removal of phenol and p-chlorophenol in enzyme reactor: horseradish peroxidase immobilized on magnetic beads

Horseradish peroxidase was immobilized on the magnetic poly(glycidylmethacrylate-co-methylmethacrylate) (poly(GMA-MMA)), via covalent bonding and used for the treatment of phenolic wastewater in continuous systems. For this purposes, horseradish peroxidase (HRP) was covalently immobilized onto magnetic poly(GMA-MMA) beds using glutaraldehyde (GA) as a coupling agent. The maximum HRP immobilization capacity of the magnetic poly(GMA-MMA)-GA beads was 3.35 mg g(-1). The immobilized HRP retained 79% of the activity of the free HRP used for immobilization. The immobilized HRP was used for the removal of phenol and p-chlorophenol via polymerization of dissolved phenols in the presence of hydrogen peroxide (H(2)O(2)). The effect of pH and temperature on the phenol oxidation rate was investigated. The results were compared with the free HRP, which showed that the optimum pH value for the immobilized HRP is similar to that for the free HRP. The optimum pH value for free and immobilized HRP was observed at pH 7.0. The optimum temperature for phenols oxidation with immobilized HRP was between 25 and 35 degrees C and the immobilized HRP has more resistance to temperature inactivation than that of the free form. Finally, the immobilized HRP was operated in a magnetically stabilized fluidized bed reactor, and phenols were successfully removed in the enzyme reactor.     

An aligned fibrous and thermosensitive hyaluronic acid-puramatrix interpenetrating polymer network hydrogel with mechanical properties adjusted for neural tissue

Journal of Materials Science - Central nervous system (CNS) injuries such as stroke or trauma can lead to long-lasting disability, and there is no currently accepted treatment to regenerate...     

有机生色团掺杂有机／无机复合基质非线性光学材料的制备与性能

以4-[乙基-（2-羟乙基）胺]-4’-硝基偶氮苯（DRl）为生色团，甲基丙烯酸甲酯（MMA）和正硅酸乙酯frEOS）为先体，通过原位溶胶-凝胶技术制备了新型的以PMMA／SiO2复合材料为掺杂基质的二阶非线性光学杂化材料，并运用提拉法在洁净的ITO导电玻璃上制备出均匀、透明的厚度为1μm左右的杂化薄膜。DSC测得杂化材料的玻璃化转变温度为130℃；二次谐波产生（SHG）测量结果表明杂化薄膜在电晕极化后二阶非线性系数d33为1．02×10^7esu；用UV-vis光谱对杂化膜在极化前后的取向和取向稳定性进行的研究表明此材料具有较高的取向度，且常温下的取向稳定性也较好，60d后取向序参数φ仍有初始值的85％。     

Mesoscopic and submicroscopic patterning in thin polymer films: Impact of the solvent

Formation of mesoscopic and submicrometric self-assembled patterns in polystyrene and polycarbonate films produced by a fast dip-coating process was studied. Mixtures of chloroform and dichloromethane were used as a solvent. The mixture's composition plays a decisive role in the patterning. Close-packed honeycomb structures comprised of 200–2000 nm pores dispersed in polymer matrix were obtained. The mechanism of self-assembling is discussed.     

Integration of polymer waveguides for optical detection in microfabricated chemical analysis systems

Multimode polymer waveguides and fiber-to-waveguide couplers have been integrated with microfluidic channels by use of a single-mask-step procedure, which ensured self-alignment between the optics and the fluidics and allowed a fabrication and packaging time of only one day. Three fabrication procedures for obtaining hermetically sealed channels were investigated, and the spectrally resolved propagation loss (400-900 nm) of the integrated waveguides was determined for all three procedures. Two chemical absorbance cells with optical path lengths of 100 and 1000 microm were furthermore fabricated and characterized in terms of coupling loss, sensitivity, and limit of detection for measurements of the dye bromothymol blue.