Sonochemical synthesis of (3-aminopropyl)triethoxysilane-modified monodispersed silica nanoparticles for protein immobilization

3-Aminopropyltriethoxysilane modified monodispersed silica nanoparticles were synthesized by a rapid sonochemical co-condensation synthesis procedure. The chemical nature of surface organic modifier on the obtained modified silica nanoparticle was characterized by ¹³C and ²⁹Si MAS Nuclear Magnetic Resonance (NMR) spectroscopies, Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA)- differential scanning calorimetry (DSC). Due to the strengthened positive surface charge of the silica nanoparticles by the modification with aminopropyl groups, the capability for bovine serum albumin (BSA) adsorption was significantly increased as compared with bare silica nanoparticles. 80 mg/g BSA was adsorbed on modified silica nanoparticles, whereas only 20 mg/g BSA could be loaded on pure silica nanoparticles. The enhanced positive surface charge repelled proteins with net positive charge and the modified silica nanoparticles exhibited negligible adsorption of lysozyme, thus a selective adsorption of proteins could be achieved.     

单分散球形介孔SiO2的合成与表征

以十六烷基三甲基氯化铵(CTAC)为模板剂,正硅酸乙酯(TEOS)为硅源,甲醇为共溶剂,氢氧化钠为催化剂,采用水热法制备出不同粒径、单一分散、高度有序的介孔SiO2微球,并利用扫描电子显微镜(SEM)对所制备的微球进行表征。考察了体系中CTAC浓度、TEOS浓度、甲醇的比率和温度等条件对所制备的SiO2微球的粒径及分散性的影响。同时采用XRD、BET、HRTEM等手段对样品进行分析表征。结果表明合成的单分散球形SiO2具有有序的六方相介孔结构,并且具有较高的比表面积。     

Trace analysis of DNA: preconcentration, separation, and electrochemical detection in microchip electrophoresis using Au nanoparticles

We have developed a simple and sensitive on-chip preconcentration, separation, and electrochemical detection (ED) method for trace analysis of DNA. The microchip comprised of three parallel channels: the first two are for the field-amplified sample stacking and subsequent field-amplified sampled injection steps, while the third one is for the microchip gel electrophoresis (MGE) with ED (MGE-ED). To improve preconcentration and separation performances of the method, the stacking and separation buffers containing the hydroxypropyl cellulose (HPC) matrix were modified with gold nanoparticles (AuNPs). The formation of AuNPs and HPC/AuNP-modified buffers were characterized by UV-visible spectroscopy and TEM experiments. The conducting polymer-modified electrode was also modified with AuNPs to enhance detection performances of the electrode. The conducting polymer/AuNP layers act as electrocatalysts for the direct detection of DNA based on their oxidation in a solution phase. The total sensitivity was improved by approximately 25 000-fold when compared with a conventional MGE-ED analysis. The calibration plots were linear (r2 = 0.9993) within the range of 0.003-1.0 pg/microL for a 20-bp DNA sample. The sensitivity was 0.20 nA/(fg/microL), with a detection limit of 5.7 amol in a 50-microL sample, based on S/N = 3. The applicability of the method for the analysis of 13 fragments present in a 100-bp DNA ladder was successfully demonstrated.     

Separation of DNA restriction fragments by capillary electrophoresis using coated fused silica capillaries.

The abilities of several different capillary electrophoresis techniques to separate DNA restriction fragments up to 23,000 bp were investigated. Methods employing electroosmotic flow in an untreated silica capillary were found to provide, at best, only partial resolution of the 23 fragments in a 1-kbp DNA ladder. By coating the inner walls of a silica capillary with poly(acrylamide) and filling these capillaries with buffers containing methylcellulose as a sleving medium, all fragments in the 1-kbp DNA ladder were separated. In addition, this technique facilitated the separation of the very large fragments in a lambda DNA-HindIII digest. Optimum resolution was obtained at low separation potentials using buffers containing at least 0.5% methylcellulose. The performance of this technique, i.e., resolution and quantitation, make capillary electrophoresis a powerful complement to slab gel electrophoresis and may make it a preferred alternative to both agarose gel electrophoresis and HPLC for applications such as the confirmation of plasmid integrity.     

Preparation of monodispersed mesoporous silica spheres with tunable pore size and pore-size effects on adsorption of Au nanoparticles and urease

Monodispersed mesoporous silica spheres (MMSS) with controllable porosity and pore size were successfully prepared by calcination method in the presence of complex salts. The effect of calcination temperature on the pore size of MMSS was examined. The results show that the pore size of MMSS samples can be tuned in the range from 3.20 to 46.80 nm by varying the calcination temperature. It is worth mentioning that the pore size of MMSS can be controlled on a much larger scale by this method compared to the templating approach, by which the pore size can only be expanded up to 10 nm. It is very advantageous for the application in loading enzymes. Moreover, it could be found that the method is feasible, effective and simple. In addition, the use of various MMSS samples as adsorbents for Au nanoparticles of different sizes as well as urease has also been demonstrated. It was confirmed that MMSS with adequate surface charge and optimum matching pore size showed excellent adsorption properties for Au nanoparticles and urease.     

Size-controlled synthesis of monodispersed gold nanoparticles stabilized by polyelectrolyte-functionalized ionic liquid

The size-controlled synthesis of monodispersed gold nanoparticles (AuNPs) stabilized by polyelectrolyte-functionalized ionic liquid (PFIL) is described. The resulting AuNPs' size, with a narrow distribution, can be tuned by the concentration of HAuCl(4). Such PFIL-stabilized AuNPs (PFIL-AuNPs) showed a high stability in water at room temperature for at least one month; they were also quite stable in solutions of pH?7-13 and high concentration of NaCl. In addition, the PFIL-AuNPs exhibited obvious electrocatalytical activity toward β-nicotinamide adenine dinucleotide (NADH for short, a cofactor in enzymatic reactions of NAD(+)/NADH(-)-dependent dehydrogenases) oxidation, suggesting a potential application for bioelectroanalysis.     

Mesoporous silica nanoparticles deliver DNA and chemicals into plants

Surface-functionalized silica nanoparticles can deliver DNA and drugs into animal cells and tissues. However, their use in plants is limited by the cell wall present in plant cells. Here we show a honeycomb mesoporous silica nanoparticle (MSN) system with 3-nm pores that can transport DNA and chemicals into isolated plant cells and intact leaves. We loaded the MSN with the gene and its chemical inducer and capped the ends with gold nanoparticles to keep the molecules from leaching out. Uncapping the gold nanoparticles released the chemicals and triggered gene expression in the plants under controlled-release conditions. Further developments such as pore enlargement and multifunctionalization of these MSNs may offer new possibilities in target-specific delivery of proteins, nucleotides and chemicals in plant biotechnology.     

High-efficiency DNA separation by capillary electrophoresis in a polymer solution with ultralow viscosity

The viscosities of some polymer solutions for DNA separation in capillary electrophoresis are generally very high, which makes them hard to pump into the capillaries. We have developed a novel sieving buffer, based on low-molecular-weight hydroxypropylmethylcellulose, to separate DNA fragments. The viscosity of this sieving matrix was at least 1 order of magnitude lower than that of traditional buffers with similar sieving effect. The influence of additives such as urea and mannitol was investigated. It was found that the double-stranded DNA (ds DNA) fragments began to denature in 3.5 M urea, and 7 M urea can denature the ds DNA completely. The presence of mannitol will decrease the overlap threshold of the polymer solution (the concentration at which the polymer molecules begin to entangle with each other), which makes it possible to separate DNA fragments in a polymer solution of relatively low concentration. The influence of the electrical field was also investigated, and it was found that the mobility of DNA fragments up to 2000 bp in length did not change greatly with different electric fields. This phenomenon implies that the DNA fragments at this range do not change their conformation with the increase of electric field as was previously believed. The possible mechanism for the separation of DNA fragments is also discussed.     

Fluorescent detection of ATP based on signaling DNA aptamer attached silica nanoparticles

Novel methods for rapid, sensitive and low-cost biomolecule detection have attracted particular interest because of their wide use in medical diagnostics, food inspection and biomedical research applications. In this work, we report a simple and efficient silica nanoparticle (NP)-based fluorescent assay for ATP detection. It takes advantage of the washing and separation properties of NPs and the structure-switch property of DNA aptamers, resulting in fluorescence change of the supernatant in the presence of targets. A linear response for ATP detection was observed from 0 to 6?mM with a detection limit of ～34?μM. This detection strategy could be generalized to other aptamer-based detection systems.     

A 265-base DNA sequencing read by capillary electrophoresis with no separation matrix

Electrophoretic DNA sequencing without a polymer matrix is currently possible only with the use of some kind of "drag-tag" as a mobility modifier. In free-solution conjugate electrophoresis (FSCE), a drag-tag attached to each DNA fragment breaks linear charge-to-friction scaling, enabling size-based separation in aqueous buffer alone. Here we report a 265-base read for free-solution DNA sequencing by capillary electrophoresis using a random-coil protein drag-tag of unprecedented length and purity. We identified certain methods of protein expression and purification that allow the production of highly monodisperse drag-tags as long as 516 amino acids, which are almost charge neutral (+1 to +6) and yet highly water-soluble. Using a four-color LIF detector, 265 bases could be read in 30 min with a 267-amino acid drag-tag, on par with the average read of current next-gen sequencing systems. New types of multichannel systems that allow much higher throughput electrophoretic sequencing should be much more accessible in the absence of a requirement for viscous separation matrix.     

Antibacterial activity of copper monodispersed nanoparticles into sepiolite

Copper monodispersed nanoparticles (2–5 nm) embedded into submicron particles of sepiolite (Mg₈Si₁₂O₃₀(OH)₄(H₂O)₄·8H₂O), suitable to be used for biological applications have been obtained after a specific treatment and subsequent reduction process. Cu/Sepiolite particles have revealed as a strong bactericide (similar to Triclosan) so that they were able to decrease the starting microorganism concentrations of Staphylococcus aureus or Escherichia coli by 99.9%.     

Effect of silica nanoparticles on compressive properties of an epoxy polymer

The effect of nanosilica on compressive properties of an Epikote 828 epoxy at room temperature was studied. A 40 wt% nanosilica/epoxy masterbatch (nanopox F400) was used to prepare a series of epoxy based nanocomposites with 5–25 wt% nanosilica content. Static uniaxial compression tests were conducted on cubic and cylindrical specimens to study the compressive stress–strain response, failure mechanisms and damage characteristics of the pure and nanomodified epoxy. It was found that the compressive stiffness and strength were improved with increasing nanosilica content without significant reduction in failure strain. The presence of nanosilica improved ductility and promoted higher plastic hardening behaviour after yielding in comparison with the unmodified resin system. This result suggested that nanoparticles introduced additional mechanisms of energy absorption to enhance the compressive properties without reducing the deformation to failure.     

Improving the signal sensitivity and photostability of DNA hybridizations on microarrays by using dye-doped core-shell silica nanoparticles

The development of new highly sensitive and selective methods for microarray-based analysis is a great challenge because, for many bioassays, the amount of genetic material available for analysis is extremely limited. Currently, imaging and detection of DNA microarrays are based primarily on the use of organic dyes. To overcome the problems of photobleaching and low signal intensities of organic dyes, we developed a new class of silica core-shell nanoparticles that encapsulated with cyanine dyes and applied the dye-doped nanoparticles as labeling in the DNA microarray-based bioanalysis. The developed nanoparticles have core-shell structure containing 15-nm Au colloidal cores with 95 dye-alkanethiol (dT)20 oligomers chemisorbed on the each Au particle surface and 10-15-nm silica coatings bearing thiol functional groups. To be utilized for microarray detection, the dye-doped nanoparticles were conjugated with DNA signaling probes by using heterobifunctional cross-linker. The prepared nanoparticle conjugates are stable in both aqueous electrolytes and organic solvents. Two-color DNA microarray-based detection was demonstrated in this work by using Cy3- and Cy5-doped nanoparticles in sandwich hybridization. The use of the fluorophore-doped nanoparticles in high-throughput microarray detection reveals higher sensitivity with a detection limit of 1 pM for target DNA in sandwich hybridization and greater photostable signals than the direct use of organic fluorophore as labeling. A wide dynamic range of approximately 4 orders of magnitude was also found when the dye-doped nanoparticles were applied in microarray-based DNA bioanalysis. In addition, the use of these dye-doped nanoparticles as the labeling in hybridization also improved the differentiation of single-nucleotide polymorphisms. This work offers promising prospects for applying dye-doped nanoparticles as labeling for gene profiling based on DNA microarray technology.     

Effect of amino groups of mesoporous silica nanoparticles on CpG oligodexynucleotide delivery

Abstract

In this study, we proposed to modify mesoporous silica nanoparticles (MSNs) with 3-aminopropyltriethoxysilane (NH₂-TES), aminoethylaminopropyltriethoxysilane (2NH₂-TES) and 3-[2-(2-aminoethylamino)ethylamino] propyl-trimethoxysilane (3NH₂-TES) for binding of cytosine-phosphate-guanosine oligodexynucleotides (CpG ODN), and investigated the effect of different amino groups of MSNs on the CpG ODN delivery. Serum stability, in vitro cytotoxicity, and cytokine interleukin-6 (IL-6) induction by MSN-NH₂/CpG, MSN-2NH₂/CpG and MSN-3NH₂/CpG complexes were investigated in detail. The results showed that three kinds of aminated-MSN-based CpG ODN delivery systems had no cytotoxicity to RAW264.7 cells, and binding of CpG ODN to MSN-NH₂, MSN-2NH₂ and MSN-3NH₂ nanoparticles enhanced the serum stability of CpG ODN due to protection by the nanoparticles. However, three aminated MSN-based CpG ODN delivery systems exhibited different CpG ODN delivery efficiency, and MSN-NH₂/CpG complexes had the highest ability to induce IL-6 secretion.     

High-resolution separation of graphene oxide by capillary electrophoresis

Separation and purification of graphene oxide (GO) prepared from chemical oxidation of flake graphite and ultrasonication by capillary electrophoresis (CE) was demonstrated. CE showed the ability to provide high-resolution separations of GO fractionations with baseline separation. The GO fractionations after CE were collected for Raman spectroscopy, atomic force microscopy, and transmission electron microscopy characterizations. GO nanoparticles (unexfoliated GO) or stacked GO sheets migrated toward the anode, while the thin-layer GO sheets migrated toward the cathode. Therefore, CE has to be performed twice with a reversed electric field to achieve a full separation of GO. This separation method was suggested to be based on the surface charge of the GO sheets, and a separation model was proposed. This study might be valuable for fabrication of GO or graphene micro- or nanodevices with controlled thickness.     

Direct synthesis of monodispersed ZnO nanoparticles in an aqueous solution

Monodispersed ZnO nanoparticles with mesopores were successfully prepared via a simple route through the transformation of Zn(NH₃)₄²⁺ precursor in the presence of sodium oleate and hydrazine at 80 °C with the pH of 8.5. Hydrazine and sodium oleate were used to control the size at 30-60 nm and to improve dispersion properties of ZnO nanoparticles. The samples were characterized by TEM, XRD, IR and TG-DTA, and the results suggest that the grains are composed of ZnO and a small quantity of oleate. The oleate plays an important role in preventing the ZnO nanoparticles from aggregating.     

单分散纳米TiO2的合成及表征

以钛酸四丁酯为原料和三乙醇胺为形态控制剂,采用溶胶-凝胶-水热法可以简单快速地合成单分散的TiO2纳米粒子.研究了反应物料配比、凝胶形成温度、水热处理温度等反应条件对粒子粒径及分散性的影响.采用XRD和TEM等手段对粒子的结构与形貌进行了表征.结果表明,三乙醇胺与钛酸四丁酯物质的量配比为2:1,凝胶形成温度为100℃,水热处理温度为140℃时,所合成的TiO2粒子分散性最好.     

Shape changes of Pt nanoparticles induced by deposition on mesoporous silica

Polyvinylpyrollidone (PVP)-capped platinum nanoparticles (NPs) are found to change shape from spherical to flat when deposited on mesoporous silica substrates (SBA-15). Transmission electron microscopy (TEM), small-angle X-ray scattering (SAXS), and extended X-ray absorption fine structure (EXAFS) analyses are used in these studies. The SAXS results indicate that, after deposition, the 2 nm NPs have an average gyration radius 22% larger than in solution, while the EXAFS measurements indicate a decrease in first neighbor co-ordination number from 9.3 to 7.4. The deformation of these small capped NPs is attributed to interactions with the surface of the SBA-15 support, as evidenced by X-ray absorption near-edge structure (XANES).     

Adsorption of avermectin on porous hollow silica nanoparticles by supercritical technology

Porous hollow silica nanoparticles with O.D. of approximately 100 nm and a wall thickness of approximately 10 nm were prepared by using inorganic CaCO3 templates. The produced PHSN were employed as a novel carrier to study the adsorption of avermectin in supercritical carbon dioxide by applying different adsorption pressure, adsorption temperature, adsorption time and volume of cosolvent. The results indicated that while increasing adsorption pressure and time always showed a positive effect on the avermectin adsorption until adsorption saturation is reached, both the adsorption temperature and volume of cosolvent require an optimal value for achieving the maximum adsorption. It was found that the optimal adsorption could be obtained at an adsorption pressure of 15 MPa and an adsorption temperature of 313 K for 90 minutes with 5 ml cosolvent. In addition, the desorption behavior of avermectin from the avermectin-loaded PHSN samples showed a sustained style: approximately 60% of avermectin was released in the first 20 minutes, while the other 40% followed a typical sustained desorption pattern and was dissolved out slowly for a time period of 3000 minutes, which is different from the quick and complete desorption from solid SiO2 carriers.