Preparation of Functional Fluorescent Microspheres Used for HTS and Their Interaction with Biomolecules

There is considerable interest in protein adsorption onto microspheres because of its importance in a wide range of biomedical applications, such as artificial tissues and organs, drug delivery systems, biosensors, solid-phase immunoassays, immunomagnetic cell separation and immobilized enzymes or catalyst. It has been well known that the interaction between proteins and microspheres plays important roles in this process. Major interaction involved in the adsorption can be classified as electrostatic, hydrophobic and hydrogen-bonding. Indeed, adsorption of proteins onto microspheres is a complex process and often can involve many dynamic steps, from the initial attachment of the protein on the surface of microspheres to the equilibrium. Also the conformation of proteins probably occurs to a certain degree of deformation or structural change due to the large area of contact. Recently, much interest has been shown in sulfonated microspheres, since sulfonate-group itself is one of components in bio-bodies, as well as is sensitive to the change of pH or ionic strength. Indeed, so far, scanty investigations have been performed in the full range. Also few researches have involved the data on adsorption rate and the maximum amount of protein adsorbed, or the reversibility of the process and conformational change of protein adsorbed as well.In present study, BSA (bovine serum albumin) was chosen as the model protein and sulfonated PMMA [poly(methyl methacrylate)] microspheres as the matrix to investigate the adsorption process.The purpose is to show some information especially the intrinsic information involved by the adsorption process Adsorption of BSA onto sulfonated microspheres (MS) has been investigated as a function of time, protein concentration and pH. The adsorption appears to be a reversible process and the presence of sulfonate groups can play important roles in the adsorption process, so as to increase the amount of protein adsorbed and influences the interaction of BSA molecules. Fig. 1 also shows that the reciprocation between unadsorbed and adsorbed BSA or rearrangement of adsorbed BSA molecules does not produce visible change in the properties of the adsorbed protein. Close to the isoelectric point of BSA (pI 4.7), the amount of protein adsorbed exhibits a maximum. A higher or lower pH results in the significant decrease of the adsorption amount. This is related to the dependence of BSA conformations at different pH conditions.     

A Novel Model for Protein Immobilization on Microspheres

The immobilization of proteins, especially receptor proteins commonly used in high through-put screening of drugs (HTS), have received great attention in recent years. There are many successful isothermal models for describing the adsorption of protein onto solid surface, such as Langmuir model, Bi-Langmuir model, Fowler model, Freundlich model, Freundlich-Langmuir model and Tekmin model etc. In all these models, Langmuir model was the most favorable one model accepted by many researchers, but the experimental results showed that it was not entirely fit to all adsorption behaviors. So new models were required for describing protein adsorption onto microspheres in different conditions.In our research, a novel isothermal model, including Langmuir and other adsorbing behaviors was presented basing on the holding degree of surface active sites and the interaction styles of protein immobilization. In Langmuir model, the adsorbing amount of protein was described as [PS] =Km[P]/1 + K[P], where [PS] was the concentration of adsorbed protein, [P] was the concentration of freeprotein at equilibrium state, and Km and K was constant. According to the interactions of protein and ligands, there were three patterns in the interactions of protein and ligands. On the similar assumption that the interaction of protein and microspheres were three styles, and based on the definition of the holding degree of surface active sites (Y), three adsorption behaviors could be described as Y K[ P ]φ/ K[P]φ+1 or ln K + φ ln[P] =ln(Y/1-Y) in which [P] was the concentration of free protein at equilibrium state, and φ and K was constant. Different scale of φ presented different adsorption behaviors, especially when φ was 1, the adsorption behavior was Langmuir adsorbing model. Figure I indicated the different adsorbing results in different adsorption behaviors (φ＞1, φ＜1,and φ=1).     

Synthesis of Cationic PEM Emulsion and Application in Waste Water Treatment

Cationic polymer as a kind of flocculant is widely applied in purification treatment of waste water. Because it has positive charge group, it is able to connect strongly the suspended matters, short cellulose and other microparticles. The research on synthesis of cationic polymer and application in treatment of waste water is very universal abroad. But domestic research on those is not general. The technology of synthesis of PEM is simple, and the production cost is low. It is easy to apply in treatment of waste water.Synthesis of PEM Emulsion FlocculantSome distilled water, PVA(poly(vinyl alcohol)), EA(ethyl acrylate), and K2S2O8(potassium persulfate) were put into reaction vessel. Kept stirring up under nitrogen. When heated the solution to 40℃, dropped the water solution of MTA[(2-methacryloxylethyl)trimethyl ammonium].Maintained the temperature at 70℃, reacted about 7-8 hours. Then got the PEM emulsion. Changed the ratio of EA and MTA. Obtained a series of PEM emulsions.Stability and Convertibility of PEM EmulsionThe test results showed that when the EA/MTA was 85/15, the PEM emulsion was most stable.When the total monomer quantity was 35%, the convertibility of PEM emulsion was the highest,i.e.98.6%.The MTA Copolymerization Ratio and Morphology of PEM EmulsionWhen the monomers EA/MTA=85/15 and total monomer quantity was 35%, the MTA copolymerization ratio of PEM emulsion was 95.15%(the highest), and the PEM emulsion was some microspheres with 100-180nm of diameter.The Test Results of PEM Emulsion in Treatment of Waste Water The PEM emulsion flocculant was applied in treatment of waste water of paper mill, and measured the precipitation time(t) and transmittancy(T). The test results were showed in table 1. The optimum value of PEM which was able to make the waste water of paper mill into clear water was 0.008%.     

CATALYSIS OF POLYSTYRENE N-HYDROXYL SULFONAMIDE FOR ESTERIFICATION OF BUTANOL WITH ACETIC ANHYDRIDE

1. INTRODUCTION The development of efficient polymer-supported catalysts has attracted much attention [1]. For obtaining polymeric catalysts, catalytically active groups were introduced onto polymers mostly by copolymerization of the appropriate monomers bearing the desired catalyticfunctionalities (e.g. imidazole, OH, and COOH) or by modification of preformed polymers.Another possibility involves the attachment of side chains, containing the desired arrangement of functional groups, o…     

The Research on Polymer Microcapsulation for Cell Technology

Microcapsulation is a technology that enwrapped the solid or liquid or some gas matter with membrane materials to form microparticles(i.e.microcapsules). The materials of microcapsule is composed of naturnal polymers or modified naturnal polymers or synthesized polymers. The water-soluble core matter can only use oil-soluble wall materials, and vice versa.Synthesized methods of polymer microcapsulesSynthesized methods with monomers as raw materialsThis kind of methods include suspension polymerization, emulsion polymerization, dispersal polymerization, precipitation polymerization,suspension condensation polymerization, dispersal condensation polymerization, deposition condensation polymerization, interface condensation polymerization, and so on.Synthesized methods with polymers as raw materialsThese methods are suspension cross-linked polymerization, coacervation phase separation,extraction with solvent evaporation, polymer deposition, polymer chelation, polymer gel,solidification of melting polymer, tray-painted ways, fluidized bed ways, and so forth.Polymer materials to synthesize microcapsules2.1. Naturnal polymer materialsThe characteristics of this kind of materials are easy to form membrane, good stability and no toxicity. The polymer materials include lipids(liposome), amyloses, proteins, plant gels, waxes, etc.2.2. Modified polymer materialsThe characteristics of these materials are little toxicity, high viscidity(viscosity), soluble salt materials. But they cannot be used in water, acidic environment and high temperature environment for a long time. The materials include all kind of derivants of celluloses.2.3. Synthesized polymer materialsThe characteristics of the materials are easy to form membrane, good stability and adjustment of membrane properties. The synthesized polymer materials include degradable polymers(PLA, PGA,PLGA, PCL, PHB, PHV, PHA, PEG, PPG and the like) and indegradable polymers(PA, PMMA,PAM, PS, PVC, PB, PE, PU, PUA, PVA and otherwise).The applications of polymer microcapsules in cell technologyThe "artificial cell" is the biological active microcapsule used in biological and medical fields.The applications of cells (including transgenic cells, the same as artificial cells) technology include several aspects as follows:3.1. Microcapsulation of artificial red cell3.2. Microcapsule of artificial cell of biological enzyme3.3. Microcapsule of artificial cell of magnetic material3.4. Microcapsule of artificial cell of active carbon3.5. Microcapsule of active biological cell     

乳糖修饰壳聚糖微载体的制备及对培养肝细胞代谢活性的影响

液体石蜡作分散介质。戊二醛作交联刑，通过反相悬浮聚合制备了微米级的壳聚糖微载体．环氧氟丙烷活化后，乳糖进行修饰．用孔糖修饰的微载体进行原代大鼠肝细胞培养，利用相差显微镜对培养细胞进行形态观察，并测定肝细胞的代谢活性，结果显示，乳糖修饰壳聚糖微载体是一种优良的肝细胞培养支架．     

Kinetic Studies on the Reaction of Sodium Hexa-cyanoferrate(II) Complex with Peroxydisulfate Anion

Introduction In the previous studies on the oxidation reaction,peroxydisulfate was widely used as an oxidizingagent.1-5 One of the advantages of this oxidant lies in itsstability in a wide range of pH values. The reaction be-tween Fe(CN)5L3- (L=N-aromatic heterocyclic li-gands) and S2O8 2- has been proved to proceed throughan outer-sphere electron transfer mechanism.5 For a re-action [(Eq. (1)] to be under an outer-sphere mechanismthe steps involved are the formation of a reactant …     

Self-assembly of Carboxyl Functionalized Polystyrene Nanospheres into Close-packed Monolayers via Chemical Adsorption

The polyacrylic acid functionalized polystyrene nanospheres were synthesized and self-assembled into irregular, densely packed monolayers in non-aqueous media. The polymer nanoparticles were chemically adhered to substrates. The morphologies of the resulting films were investigated. The impact of the volume fraction of alcohol in the mixed solvents on the particle adsorption and fabrication of nanosphere assembled films was examined.     

Microwave Synthesis, Characterization and DNA-binding Properties of a New Cobalt（ Ⅱ ） Complex with 2,6-Bis（benzimidazol-2-yl）pyridine

A new cobalt(Ⅱ) complex with tridentate ligand 2, 6-bis (benzimidazol-2-yl)pyridine has been synthesized by microwave irradiation method and characterized by elemental analysis,electrochemical and spectral methods. The binding of the complex with calf thymus DNA has also been investigated by absorption and fluorescence spectra.     

Synthesis and Photochemical Properties of a New Substituted Phenol Covalently Linked to Ruthenium （Ⅱ） Tris-bipyridine Containing Four Ester Groups as Sensitizer

As photosensitizer for solar cell, a new ruthenium (Ⅱ) complex with four ester groups had been synthesized, in which a phenol substituted by {[(2-hydroxy-5-tert-butylbenzyl)(pyridyl-2-methyl)amino]methyl} is covalently linked to ruthenium (Ⅱ) tris-bipyridine. The structures of the new compounds were confirmed by NMR and ESI-MS spectra. The electrochemical and photochemical properties were also studied.