Preparation of bovine serum albumin surface-imprinted submicrometer particles with magnetic susceptibility through core-shell miniemulsion polymerization

Molecular imprinting is a state-of-the-art technique for preparing mimics of natural, biological receptors. Nevertheless, the imprinting of macromolecules like proteins remains a challenge due to their bulkiness and sensitivity to denaturation. In this work, a surface imprinting strategy based on covalently immobilized template molecules was adopted for protein imprinting. Bovine serum albumin (BSA) surface-imprinted submicrometer particles (500-600 nm) with magnetic susceptibility were prepared through a two-stage core-shell miniemulsion polymerization system using methyl methacrylate and ethylene glycol dimethacrylate as functional and cross-linking monomers, respectively. The particles possessed a novel red blood cell-like structure and exhibited a very favorable recognition property toward the template BSA molecules in aqueous medium. In a two-protein system, the particles had shown a very high specific recognition of the template proteins over the nontemplate proteins. The magnetic susceptibility was imparted through the successful encapsulation of Fe3O4 nanoparticles. Their superparamagnetic nature increases their potential applications in the fields such as magnetic bioseparation, cell labeling, and bioimaging. In addition, the importance of template immobilization for successful protein imprinting had also been illustrated to demonstrate the potential of this approach as a general methodology for protein imprinting.     

超顺磁性Fe3O4纳米颗粒的合成及应用

总结了超顺磁性Fe3O4纳米颗粒的常用制备方法:沉淀法、水热法、微乳液法、模板合成法及生物矿化合成法,并综述了其研究现状,同时比较了它们各自的优缺点及所面临的问题.此外,还概述了超顺磁性Fe3O4纳米颗粒在靶向药物、癌症治疗、磁共振成像(MR I)及生物活性物质的检测和分离等生物医学方面的应用,并对前景进行了展望.     

Preparation of superparamagnetic ribonuclease A surface-imprinted submicrometer particles for protein recognition in aqueous media

Superparamagnetic ribonuclease A surface-imprinted polymeric particles that can preferentially bind the template protein in an aqueous environment were prepared in this study. Methyl methacrylate and ethylene glycol dimethacrylate were employed as the functional and cross-linker monomers, respectively. Regularly shaped submicrometer (700-800 nm) particles imprinted with ribonuclease A were successfully prepared using redox-initiated miniemulsion polymerization. Nanosized Fe3O4 magnetite was encapsulated in the imprinted particles with good encapsulation efficiency (17.5 wt %) for the incorporation of the superparamagnetic property. Good selectivity toward the template over the control protein in an aqueous environment was demonstrated by the imprinted particles in the batchwise and competitive rebinding tests with the highest template loading, Qmax, of 127.7 mg/g observed in the batch rebinding test. Given the small sizes of the imprinted particles and the presence of the binding sites on the surface, the rebinding process was kinetically favorable despite the sheer bulk of the macromolecules. In the desorption study, it was found that the more hydrophobic solvent was more effective for ribonuclease A desorption from the imprinted particles. This indicated that the hydrophobic effect was probably the main form of interaction responsible for the template rebinding to the imprinted sites in an aqueous media.     

L-histidine imprinted synthetic receptor for biochromatography applications

We have proposed novel surface-imprinted beads for selective separation of cytochrome c (cyt c) by N-methacryloyl-(L)-histidine-copper(II) [MAH-Cu(II)] as a new metal-chelating monomer via metal coordination interactions and histidine template. We have combined molecular imprinting with the ability of histidine to chelate metal ions to create ligand exchange beads suitable for the binding of cyt c (surface histidine exposed protein). The histidine imprinted beads were produced by suspension polymerization of MAH-Cu(II)-L-histidine and ethylene glycol dimethacrylate. After polymerization, the template (L-histidine) was removed from the beads using methanolic KOH, thus getting histidine imprinted metal-chelate beads. L-Histidine imprinted metal-chelate beads can be used several times without considerable loss of cyt c adsorption capacity. The association constant (Ka) for the specific interaction between the template imprinted polymer and the template (L-histidine) itself were determined by Scatchard plots using L-histidine imprinted beads and found as 58,300 M(-1). Finally, we have used these histidine imprinted beads for cyt c and ribonuclease A (surface histidine exposed proteins) and enantiometric separation of D- and L-histidine by FPLC.     

Stimulus-responsive fluidic dispersions of rod shaped liquid crystal polymer colloids

Electrically responsive liquid crystal polymer nanorods were fabricated by template synthesis. Liquid crystal monomers are templated by alumina membranes. Molecular ordering of the liquid crystal molecules resulted from the confinement in the sub-micron channels and this ordering can be captured permanently through photo-polymerization. Template removal and sonication result in individual rods that can be reoriented by applied electrical and magnetic fields. Such anisotropic particles have significant potential applications in electro-rheological fluids and in active mixing in microfluidic channels.     

Surface-imprinted core-shell nanoparticles for sorbent assays

In this paper, we present a general protocol for the making of surface-imprinted core-shell nanoparticles via surface reversible addition-fragmentation chain-transfer (RAFT) polymerization using RAFT agent functionalized model silica nanoparticles as the chain-transfer agent. In this protocol, trichloro(4-chloromethylphenyl)silane was immobilized on the surface of SiO2 nanoparticles, forming chloromethylphenyl functionalized silica (silica-Cl). RAFT agent functionalized silica was subsequently produced by substitute reaction of silica-Cl with PhC(S)SMgBr. The grafting copolymerization of 4-vinylpyridine and ethylene glycol dimethacrylate using surface RAFT polymerization and in the presence of 2,4-dichlorophenoxyacetic acid as the template led to the formation of surface-imprinted core-shell nanoparticles. The resulting surface-imprinted core-shell nanoparticles bind the original template 2,4-D with an appreciable selectivity over structurally related compounds. The potential use of the surface-imprinted core-shell nanoparticles as the recognition element in the competitive fluorescent binding assay for 2,4-D was also demonstrated.     

超顺磁性羟基磷灰石/壳聚糖棒材的制备

以壳聚糖膜为模板将磁性羟基磷灰石前驱体的壳聚糖醋酸溶液与NaOH凝固液隔离,OH^-向磁性羟基磷灰石前驱体的壳聚糖醋酸溶液内部的渗透引起pH值变化,导致质子化的壳聚糖在模板上沉积的同时无机物就地生成,原位复合制备出无机纳米颗粒分散均匀的磁性羟基磷灰石／壳聚糖（HA／CS）复合棒材．生成的超顺磁性四氧化三铁和羟基磷灰石颗粒大小均一（大约长30nm,宽20nm）,在基质中分布均匀、没有出现明显的团聚现象．     

Optimization of carbothermic synthesis of zinc-oxide micro- and nanorod arrays and their morphometric parameters

The influence of argon-buffer pressure and distance from the material source on morphometric parameters of arrays of vertically aligned ZnO nano- and microrods was studied. The rods were grown by the carbothermic method on Si(100) substrates with a ZnO thin-film sublayer. Systematic studies of catalyst-free growth, as well as of the effect of growth catalysts Au and Cu on the rods’ formation process and their sizes, were performed. It was found that a catalyst plays a role only on the initial stage and does not participate in the consequent rods’ growth under the applied synthesis conditions. The possibility of controlling the average diameters, lengths, and densities of superficial distribution of the rods within the ranges of 100–300 nm, 1.5–9 μm, and (2.8–5.3) × 10⁸ cm^?2, respectively, was demonstrated.     

Synthesis of magnetite/amphiphilic polymer composite nanoparticles as potential theragnostic agents

This study describes the synthesis of magnetite/amphiphilic polymer composite nanoparticles that can be potentially used simultaneously for cancer diagnosis and therapy. The synthesis method was a one-shot process wherein magnetite nanoparticles were mixed with core-crosslinked amphiphilic polymer (CCAP) nanoparticles, prepared using a copolymer of a urethane acrylate nonionomer (UAN) and a urethane acrylate anionomer (UAA). The CCAP nanoparticles had a hydrophobic core and a hydrophilic exterior with both PEG segments and carboxylic acid groups, wherein the magnetite nanoparticles were coordinated and stabilized. According to DLS data, the ratio of UAN to UAA and the ratio of magnetite to polymer are keys to controlling the size and thus, the stability of the composite nanoparticles. The magnetic measurement indicated that the composite nanoparticles had superparamagnetic properties and high saturation magnetization. The preliminary magnetic resonance imaging showed that the particles produced an enhanced image even when their concentration was as low as 80 microg/ml.     

Synthesis of ultrafine amorphous Fe–B alloy nanoparticles using anodic aluminum oxide templates

Ultrafine amorphous Fe–B alloy nanoparticles are self-assembled within anodic aluminum oxide templates by combining a preparation process of Fe–B nanoparticles with a template method. Scanning electron microscopy, inductively coupled plasma-atomic emission spectrometry, X-ray diffraction spectrometry, Mössbauer spectrometry, and vibrating sample magnetometry are employed to study the morphology, chemical composition, structure, and magnetic properties of the nanoparticle assemblies, respectively. The results show that the alloy particles are amorphous with a boron content of 24 at. % and can be in shape of sphere and rod by controlling the duration of preparation. There is a narrow distribution of the sizes of spherical nanoparticles with an average diameter below 35 nm in relatively short preparation time, while rods are found in longer time. The measurements of magnetic properties indicate that the nanoparticles are mostly in superparamagnetic state and the self-assembly of the nanoparticles has a weak magnetic anisotropy with an easy direction perpendicular to the template plane.     

Templated growth of superparamagnetic iron oxide nanoparticles by temperature programming in the presence of poly(vinyl alcohol)

Magnetite (Fe₃O₄) nanostructures with different morphologies including uniform nanoparticles, magnetic beads and nanorods were synthesized via a co-precipitation method. The synthesis process was performed at various temperatures in the presence of polyvinyl alcohol (PVA) at different concentrations. It is shown that small amounts of PVA act as a template in hot water (70 °C), leading to the oriented growth of Fe₃O₄ nanorods, which was confirmed by selected area electron diffraction. Individually coated magnetite nanoparticles and magnetic beads were formed at a relatively lower temperature of 30 °C in the folded polymer molecules due to the thermo-physical properties of PVA. When a moderate temperature (i.e. 50 °C) was used, nanorods and nanobeads co-existed. At higher concentrations of PVA (polymer/iron mass ratio of 5), however, the formation of magnetic beads was favored. The nanorods were shown to be unstable upon exposure to electron beams. Freezing/thawing process was applied post synthesis as temperature programming to fabricate stable nanorods with rigid walls.     

Electrospun polyvinylpyrrolidone fibers with high concentrations of ferromagnetic and superparamagnetic nanoparticles

Electrospun polymer fibers were prepared containing mixtures of different proportions of ferromagnetic and superparamagnetic nanoparticles. The magnetic properties of these fibers were then explored using a superconducting quantum interference device. Mixed superparamagnetic/ferromagnetic fibers were examined for mesoscale magnetic exchange coupling, which was not observed as theoretically predicted. This study includes some of the highest magnetic nanoparticle loadings (up to 50 wt%) and the highest magnetization values (≈ 25 emu/g) in an electrospun fiber to date and also demonstrates a novel mixed superparamagnetic/ferromagnetic system.     

Description of the process of magnetization of hollow cylindrical rods from soft magnetic materials in a homogeneous quasi-static magnetic field of a solenoid. Hysteresis loop

On the basis of the arctangential approximation of the magnetic hysteresis loop of the material and the expression for the central demagnetization coefficient of hollow cylindrical rods from soft magnetic materials, a method for calculating the hysteresis loops of these rods in a homogeneous quasi-static field of a solenoid is proposed. Calculation and comparison to experimental data of the basic parameters of the magnetic loops hysteresis of hollow cylindrical rods with a different value of the demagnetization coefficient have been made. __________ Translated from Inzhenerno-Fizicheskii Zhurnal, Vol. 80, No. 6, pp. 181–188, November–December, 2007.     

Two- and three-dimensional micro/nanostructure patterning of CdS-polymer nanocomposites with a laser interference technique and in situ synthesis

Two-?and three-dimensional (2D and 3D) micro/nanostructures of CdS-polymer nanocomposites have been successfully patterned, combining photopolymerization via a laser four-beam interference technique with in situ synthesis of CdS nanoparticles in the patterned polymer matrix. The morphology and optical properties of CdS nanoparticles in polymer matrices have been confirmed using TEM, XRD, FTIR, UV-vis absorption and fluorescence spectroscopy. Laser irradiation time and film thickness are certified to be the key factors for the control of the micro/nanostructures. With thickening film, the fabricated microstructures of CdS-polymer nanocomposites were dramatically changed from 2D rods to 3D networks which were composed of nanofibres, nanometre-scale walls and micrometre-scale rods. These kinds of 2D and 3D micro/nanostructures could be expected as potential applications in the development of nanotechnology, such as nanomedical systems, micro-fluidic chips, nanoreactors and micro/nanopurification or separation systems.     

Indium microrod tags for electrochemical detection of DNA hybridization

The preparation and advantages of indium microrod tracers for solid-state electrochemical detection of DNA hybridization are described. The cylindrical metal particles were prepared by a template-directed electrochemical synthetic route involving plating of indium into the pores of a host membrane. The linear relationship between the charge passed during the preparation and the resulting particle size allows tailoring of the sensitivity of the electrical DNA assay. The resulting micrometer-long rods thus offer a greatly lower detection limit (250 zmol), as compared to common bioassays' spherical nanoparticle tags. Indium offers a very attractive electrochemical stripping behavior and is not normally present in biological samples or reagents. Solid-state derivative-chronopotentiometric measurements of the indium tracer have been realized through a "magnetic" collection of the DNA-linked particle assembly onto a thick-film electrode transducer. Factors affecting the performance, including the preparation of the microrods and pretreatment of the transducer surface, were evaluated and optimized. The resulting protocol offers great promise for other affinity bioassays, as well as for electrical coding and identification (through the plating of different metal markers and of multimetal redox-encoded tags).     

Template-mediated synthesis and bio-functionalization of flexible lignin-based nanotubes and nanowires

Limitations of cylindrical carbon nanotubes based on the buckminsterfullerene structure as delivery vehicles for therapeutic agents include their chemical inertness, sharp edges and toxicological concerns. As an alternative, we have developed lignin-based nanotubes synthesized in a sacrificial template of commercially available alumina membranes. Lignin is a complex phenolic plant cell wall polymer that is generated as a waste product from paper mills and biorefineries that process lignocellulosic biomass into fuels and chemicals. We covalently linked isolated lignin to the inner walls of activated alumina membranes and then added layers of dehydrogenation polymer onto this base layer via a peroxidase-catalyzed reaction. By using phenolic monomers displaying different reactivities, we were able to change the thickness of the polymer layer deposited within the pores, resulting in the synthesis of nanotubes with a wall thickness of approximately 15?nm or nanowires with a nominal diameter of 200?nm. These novel nanotubes are flexible and can be bio-functionalized easily and specifically, as shown by in vitro assays with biotin and Concanavalin A. Together with their intrinsic optical properties, which can also be varied as a function of their chemical composition, these lignin-based nanotubes are expected to enable a variety of new applications including as delivery systems that can be easily localized and imaged after uptake by living cells.     

Magnetic Core/Shell and Quantum‐Confined Semiconductor Nanoparticles via Chimie Douce Organometallic Synthesis

Ligand‐stabilized metal atoms provide a unique entry to the synthesis of magnetic nanosized metal/metal oxide particles. When this technique is used in connection with a mesoporous template, formation of superparamagnetic particles in the pores of the template via a “ship‐in‐the‐bottle” technique is possible. This chimie douce approach works also for pure metal oxides, such as TiO₂. The Figure shows a sculpture found at the Trinity College campus, Dublin, Ireland representing a typical core/shell arrangement often found for composite nanoparticles.     

Polyaniline microrods synthesized by a polyoxometalates/poly(vinyl alcohol) microfibers template

Polyaniline (PAn) microrods doped with H₄SiW₁₂O₄₀ (HPA) were obtained by a H₄SiW₁₂O₄₀/Poly(vinyl alcohol) microfibers template method. The morphology of the polyaniline microrods was observed by scanning electron microscope (SEM) photograph. The average diameter of the polyaniline microrods was between 200 and 250 nm. IR spectra and X-ray diffraction characterized the structure of the polyaniline doped with H₄SiW₁₂O₄₀. The conductivity of the polyaniline microrods was measured by a four-probe method. The highest conductivity of the polyaniline microrods was 3.9 S/cm.     

Preparation of superparamagnetic and flexible γ-Fe<Subscript>2</Subscript>O<Subscript>3</Subscript> nanowire arrays in an anodic aluminum oxide template

Superparamagnetic and flexible Fe₂O₃ nanowire arrays were fabricated by the controlled electrostatic assembly of iron oxide nanoparticles and poly(dimethyldiallylammonium chloride) (PDADMAC) in an anodic aluminum oxide (AAO) template. The micrograph of iron oxide nanowire arrays was characterized by field emission scanning electron microscopy. The magnetic hysteresis loops obtained by a vibrating sample magnetometer confirm that the nanowire arrays have superparamagnetic properties. The filling ratio of iron oxide nanoparticles and polymers in the AAO template was affected by four factors, including the concentration of iron oxide nanoparticles, the pore diameter of the AAO template, the charge ratio of iron oxide nanoparticles and PDADMAC, and the molecular weight of polyacrylic acid. The effect of the AAO template on the diameter and length of the nanowire arrays was also analyzed. In addition, the nanowire arrays were shown to be flexible because of the presence of polymers. These nanowire arrays with superparamagnetic and flexural properties have potential applications in sensor probes.     

Synthesis of water-compatible surface-imprinted composite microspheres with core–shell structure for selective recognition of thymopentin from aqueous solution

A novel water-compatible surface-imprinted core–shell microsphere, which had multiple non–covalent interactions with template molecule, was successfully prepared by the surface grafting polymerization method in acetonitrile–water systems with thymopentin as template through ionic liquid-functionalized polyethyleneglycolmethacrylate-co-vinylimidazole microsphere as the matrix. The average diameter of matrix was 1 μm ± 20 nm and the thickness of imprinted layer was about 50 nm. The results of static adsorption experiments indicated that ionic liquid-functionalized molecularly imprinted microspheres showed the good adsorption capacity and specific recognition for template peptide. The binding-isotherm analysis showed that Langmuir isotherm models gave a good fit in the range of concentrations, suggesting that there was only one kind of binding site in imprinted layer. Measurements of the binding kinetics revealed that surface-imprinted composite microspheres reached peptide-adsorption equilibrium in 60 min and the maximum adsorption capacity for TP5 was 38.4 mg g^?1. The effects of pH, salt concentration, and temperature on the adsorption capacities were investigated. The microspheres were found to have a high specificity for TP5 with little affinity for BSA and Hb. Finally, the core–shell microspheres can be reused with only 15.6 % decrease in TP5 adsorption capacity after six times.