Immobilization of Acetylcholinesterase onto Carbon Nanotubes Utilizing Streptavidin-biotin Interaction for the Construction of Amperometric Biosensors for Pesticides

Abstract

Carbon nanotubes (CNTs) are very promising materials onto which bioactive molecules can be immobilized in the construction of biosensors. Streptavidin was used as a molecular linker to immobilize biotinylated acetylcholinesterase (AChE) on CNTs in a gentle and controllable fashion for pesticide biosensors. Glassy carbon electrodes coated with the CNT-enzyme complex had high affinity for the substrate acetylthiocholine and produced strong peak oxidation currents in electrochemical assays. We also propose a new method, i.e., the use of relative net slope rather than the percentage of inhibition, in the calculation of pesticide concentrations. The biosensors could detect low levels of the pesticide methyl paraoxon.     

Preparation and characterization of carbon nanotubes-polymer/CdSe hybrid nanocomposites through combining electrostatic adsorption and ATRP technique

This paper describes a new strategy through noncovalent functionalization of multi-walled carbon nanotubes (MWNTs) by a kind of new copolymer Polyethyleneimine-graft-Polyacrylonitrile for attaching CdSe nanoparticles onto the MWNTs to fabricate Carbon Nanotube/CdSe heterostructures. Polyethyleneimine (PEI), an amino-rich cationic polyelectrolyte, can interact with the MWNTs through electrostatic interaction. Then, CNT/PEI-g-PAN was successfully prepared by in situ atom transfer radical polymerization (ATRP), which did not introduce defects to the structure of CNTs. Thus, CdSe nanoparticles can be covalently coupled to functionalized carbon nanotubes (CNTs) in a uniform and controllable manner. Moreover, this method ensures good dispersion and high stability in any commonly used organic or inorganic solvent. In this manner, our strategy allows the attachment of various colloidal nanoparticles to CNTs, independent of their surface properties, i.e. hydrophilic or hydrophobic. TEM, XRD, EDS and FT-IR are all used to characterize the CNT/CdSe composite materials. In addition, the optical properties are investigated by UV–vis spectrum.     

A direct route toward assembly of nanoparticle-carbon nanotube composite materials

The preparation of nanocomposite materials from carbon nanotubes (CNTs) and metal or metal oxide nanoparticles has important implications to the development of advanced catalytic and sensory materials. This paper reports findings of an investigation of the preparation of nanoparticle-coated carbon nanotube composite materials. Our approach involves molecularly mediated assembly of monolayer-capped nanoparticles on multiwalled CNTs via a combination of hydrophobic and hydrogen-bonding interactions between the capping/mediating shell and the CNT surface. The advantage of this route is that it does not require tedious surface modification of CNTs. We have demonstrated its simplicity and effectiveness for assembling alkanethiolate-capped gold nanoparticles of 2-5 nm core sizes onto CNTs with controllable coverage and spatially isolated character. The loading and distribution of the nanoparticles on CNTs depend on the relative concentrations of gold nanoparticles, CNTs, and mediating or linking agents. The composite nanomaterials can be dispersed in organic solvent, and the capping/linking shells can be removed by thermal treatment to produce controllable nanocrystals on the CNT surfaces. The nanocomposite materials are characterized using transmission electron microscopy and Fourier transform infrared spectroscopy techniques. The results will be discussed in terms of developing advanced catalytic and sensory nanomaterials.     

Dispersing carbon nanotubes in aqueous solutions by a silicon surfactant: Experimental and molecular dynamics simulation study

The dispersion effect of carbon nanotubes (CNTs) in aqueous solutions by a silicon surfactant (ethoxy modified trisiloxane, named Ag-64) was investigated in detail using experimental method and molecular dynamics simulation. The Si–O–Si chain of silicon surfactant was flexible due to long Si–C bond and it could easily wrap onto the surface of CNTs through hydrophobic and other intermolecular interactions. The hydrophilic part of PEO provided the CNTs dispersed in the aqueous solution and prevented CNTs from aggregating in water through steric stabilization. It was found that Ag-64 could disperse CNTs with different diameters and it was an effective dispersing agent. The results of molecular dynamics simulation indicated that Ag-64 molecules could wrap onto the surface of CNTs leading to steric stabilization so that it could well disperse CNTs, and Van der Waals attraction was the dominating force of Ag-64 adsorbing onto CNTs. Our study may provide experimental and theoretical basis for using silicon surfactants to disperse CNTs, which can open the avenue of new applications for silicon surfactants.     

Cycloaddition reactions: a controlled approach for carbon nanotube functionalization

Controlled functionalization of carbon nanotubes (CNTs) through the use of cycloaddition reactions is described. By employing various cycloaddition reactions, a wide range of molecules could be coupled onto CNTs without disruption of the structural integrity as well as with a statistical distribution of functional groups onto the surface of the CNTs. The cycloaddition reactions represent an effective and tailored approach for preparing CNT-based advanced hybrid materials that would be useful for a wide range of applications from nanobiotechnology to nanoelectronics.     

Chemically grafting carbon nanotubes onto carbon fibers for enhancing interfacial strength in carbon fiber/HDPE composites

We propose a novel method to uniformly graft high‐density carbon nanotubes (CNTs) onto carbon fiber (CF) by using coupling agents. Coupling agents can supply much more active groups, which is beneficial for grafting high‐density CNTs onto CF surface. After CNT grafting treatment, there are still substantial amounts of reactive groups, which can further react with various types of molecules to meet different requirements. To create chemical bonding between CF and high‐density polyethylene, CF‐CNT was further grafted get reinforcement. The interfacial adhesion of the resulting composites showed a dramatic improvement.     

Surfactant-free nanofluids containing double- and single-walled carbon nanotubes functionalized by a wet-mechanochemical reaction

Single-walled carbon nanotubes (SWNTs) and double-walled carbon nanotubes (DWNTs) have been functionalized through the wet-mechanochemical reaction method. Results from the infrared spectrum and zeta potential measurements show that the hydroxyl groups have been introduced onto the treated SWNT and DWNT surfaces. Transmission electron microscope observations revealed that the SWNTs and DWNTs were cut short after being milled. SWNTs and DWNTs with optimized aspect ratio can be obtained by adjusting the ball milling parameters. Thermal conductivity enhancement of water-based nanofluids containing treated carbon nanotubes (CNTs) shows augmentation with the increase of temperature mainly due to the effects of an ordering liquid layer forming around the chemical surfaces of CNTs. Moreover, the thicker interfacial layer of water molecules on the surfaces of CNTs with smaller diameter, such as SWNTs, is in favor of greater thermal conductivity enhancement compared with the thinner one on the surfaces of DWNTs or MWNTs with larger diameter.     

Controlled Chemical Derivatisation of Carbon Nanotubes with Imaging,Targeting, and Therapeutic Capabilities

In drug delivery, carbon nanotubes (CNTs) hold a great potential as carriers because of their ability to easily cross biological barriers and be internalised into cells. Their high aspect ratio allows multi‐functionalisation and their development as a multimodal platform for targeted therapy. In this article, we report the controlled covalent derivatisation of triple‐functionalised CNTs with the anticancer drug gemcitabine, folic acid as a targeting ligand and fluorescein as a probe. The anticancer activity of gemcitabine was maintained after covalent grafting onto the CNTs. The functionalised nanotubes were internalised into both folate‐positive and negative cells, suggesting the passive diffusion of CNTs. Overall, our approach is versatile and offers a precise chemical control of the sidewall functionalisation of CNTs and the possibility to manoeuvre the types of functionalities required on the nanotubes for a multimodal therapeutic strategy.     

碳纳米管在药物和基因转运领域的研究进展

当前,国内外的许多研究小组都致力于开发出新型有效的药物和基因转运系统,用于改善多种治疗因子的药理学作用并降低其毒性。在纳米材料这一类中,碳纳米管（Carbon Nanotubes, CNTs）正逐步引起人们的关注。功能化的CNTs的两个关键优势在于它具有很强的细胞穿透能力和较低的细胞毒性,使其在药物和基因转运领域中的应用成为可能。CNTs可通过形成稳定的共价键或形成以非共价键为基础的超分子结合物来运载肽类、蛋白质、核酸和药物等活性分子,并将其运送至特定的组织、器官中以表达特殊的生物学功能。针对这一研究热点,本文综述了近几年国内外关于碳纳米管在药物和基因转运领域中的应用进展,并探讨了其毒性,以期为这一领域中的研究工作者提供参考。     

水溶液中碳纳米管的表面增强拉曼光谱研究

合成了碳纳米管和金纳米颗粒的复合物, 测量了水溶液相中复合物的表面增强拉曼光谱, 结果表明, 碳纳米管的巯基化修饰可以提高碳纳米管与金纳米颗粒复合的效率, 随着金纳米颗粒负载量的增加, 碳纳米管的拉曼信号逐渐增强. 加入己二胺分子可以减小金纳米颗粒之间的距离使表面增强效应更显著, 碳纳米管的拉曼光谱得到进一步的增强. 还可进一步在复合体系中加入对巯基苯胺和罗丹明B等小分子拉曼探针, 利用金纳米颗粒的表面增强效应, 这种多元复合体系有望作为多通道拉曼成像探针材料.     

水分散性碳纳米管的制备研究进展

采用化学修饰的方法制备水分散性碳纳米管(Carbon nanotubes,CNTs)是扩大CNTs在生物传感器、医药、功能材料和电子电器元件等领域应用的重要手段。化学修饰法主要包括非共价修饰和共价修饰两种,非共价修饰是采用表面活性剂、有共轭平面结构的分子以及聚合物或生物大分子对CNTs进行表面包覆;共价修饰包括"grafting to"和"grafting from"两种类型。本文从反应原理和实施方法上综述了化学修饰法制备水分散性CNTs的研究进展,并对水分散性CNTs的研究和应用前景进行了展望。     

Bioavailability of metallic impurities in carbon nanotubes is greatly enhanced by ultrasonication

Metallic impurities within carbon nanotubes (CNTs) are considered as the main cause of their toxicity. Ultrasonication is a common procedure used to purify and obtain homogeneous dispersions of CNTs as well as to mix them with other components for further processing into composites. Herein, the influence of ultrasonication upon the bioavailability of metallic impurities in CNTs was investigated. We showed that even ultrasonication times as short as 5?min significantly enhanced the bioavailability of metallic impurities, which can therefore interact more actively with biologically important molecules. These findings will have profound impact on the processing of CNTs as well as on nanotoxicity studies.     

Selective Synthesis of Conjugated Chiral Macrocycles: Sidewall Segments of (−)/(+)‐(12,4) Carbon Nanotubes with Strong Circularly Polarized Luminescence

Carbon nanotubes (CNTs) have unusual physical properties that are valuable for nanotechnology and electronics, but the chemical synthesis of chirality‐ and diameter‐specific CNTs and π‐conjugated CNT segments is still a great challenge. Reported here are the selective syntheses, isolations, characterizations, and photophysical properties of two novel chiral conjugated macrocycles ([4]cyclo‐2,6‐anthracene; [4]CAn_2,6 ), as (?)/(+)‐(12,4) carbon nanotube segments. These conjugated macrocyclic molecules were obtained using a bottom‐up assembly approach and subsequent reductive elimination reaction. The hoop‐shaped molecules can be directly viewed by a STM technique. In addition, chiral enantiomers with (?)/(+) helicity of the [4]CAn_2,6 were successfully isolated by HPLC. The new tubular CNT segments exhibit large absorption and photoluminescence redshifts compared to the monomer unit. The carbon enantiomers are also observed to show strong circularly polarized luminescence (g_lum≈0.1). The results reported here expand the scope of materials design for bottom‐up synthesis of chiral macrocycles and enrich existing knowledge of their optoelectronic properties.     

Selective Synthesis of Conjugated Chiral Macrocycles: Sidewall Segments of (−)/(+)-(12,4) Carbon Nanotubes with Strong Circularly Polarized Luminescence

Carbon nanotubes (CNTs) have unusual physical properties that are valuable for nanotechnology and electronics, but the chemical synthesis of chirality- and diameter-specific CNTs and π-conjugated CNT segments is still a great challenge. Reported here are the selective syntheses, isolations, characterizations, and photophysical properties of two novel chiral conjugated macrocycles ([4]cyclo-2,6-anthracene; [4]CAn_2,6 ), as (−)/(+)-(12,4) carbon nanotube segments. These conjugated macrocyclic molecules were obtained using a bottom-up assembly approach and subsequent reductive elimination reaction. The hoop-shaped molecules can be directly viewed by a STM technique. In addition, chiral enantiomers with (−)/(+) helicity of the [4]CAn_2,6 were successfully isolated by HPLC. The new tubular CNT segments exhibit large absorption and photoluminescence redshifts compared to the monomer unit. The carbon enantiomers are also observed to show strong circularly polarized luminescence (g_lum≈0.1). The results reported here expand the scope of materials design for bottom-up synthesis of chiral macrocycles and enrich existing knowledge of their optoelectronic properties.     

A simple route for the attachment of colloidal nanocrystals to noncovalently modified multiwalled carbon nanotubes

A simple strategy for the fabrication of multiwalled carbon nanotubes (MWNTs)–nanocrystal (NC) heterostructures is shown. Different nanoparticles can be covalently coupled to functionalized carbon nanotubes (CNTs) in a uniform and controllable manner. MWNTs have been functionalized by a polymer wrapping—technique that is non-invasive, and does not introduce defects to the structure of CNTs; the polymer is noncovalently adsorbed on the MWNT's surface. Moreover, this method ensures good dispersion and high stability in any commonly used organic or inorganic solvent. In this manner, our strategy allows the attachment of various colloidal nanoparticles to CNTs, independent of their surface properties, i.e. hydrophilic or hydrophobic.     

上转换纳米粒子结合DNA链式扩增技术在阿霉素光控释放中的应用

纳米技术的发展使得纳米材料可以通过不同的表面包覆和修饰而在生物医药中发挥应用。 构建简单、经济、药物释放可控的生物相容性纳米药物仍是纳米生物化学领域的重点。 我们构建的纳米载药体系(DDS)以NaYF₄：Yb/Tm上转换纳米粒子为载体,在其表面通过光致断键型小分子4,5-二甲氧基-2-硝基苯基乙酮(DMNPE)连接一段短单链DNA,利用DNA链式扩增技术(HCR)来调节纳米粒子最终修饰的双链DNA的总量,从而控制对抗癌药物阿霉素(Dox)的担载量,在980 nm激光照射下上转换纳米粒子发射可切断DMNPE连接的近紫外光,协同胞内DNA酶的作用达到对药物的可控释放。 由于近红外光照对生物组织具有较好的穿透能力,此体系能够对病灶位置有更好的光靶向性从而减少药物的毒副作用。     

Synergistically Enhanced Dispersion of Native Protein–Carbon Nanotube Conjugates by Fluoroalcohols in Aqueous Solution

Carbon nanotubes (CNTs) are anticipated as an important new material for use in nanotechnology applications because of their excellent mechanical and electrical properties. For their development, a highly stable dispersion of debundled CNTs is indispensable. Herein we present a new method to enhance dispersibility of single‐walled carbon nanotubes (SWNTs) with proteins using alcohols as co‐solvents. Addition of fluoroalcohols in solution increased the SWNT dispersion by more than one order of magnitude without protein denaturation. Enhancement of SWNT dispersion through addition of alcohols was attributed to the decreased hydrophobic interaction among SWNTs. This novel approach enables us to produce biofunctional CNTs such as one‐dimensional nanobiosensors and drug carriers that can penetrate cells.     

Conditional glycosylation in eukaryotic cells using a biocompatible chemical inducer of dimerization

Chemical inducers of dimerization (CIDs) are cell-permeable small molecules capable of dimerizing two protein targets. The most widely used CID, the natural product rapamycin and its relatives, is immunosuppressive due to interactions with endogenous targets and thus has limited utility in vivo. Here we report a new biocompatible CID, Tmp-SLF, which dimerizes E. coli DHFR and FKBP and has no endogenous mammalian targets that would lead to unwanted in vivo side effects. We employed Tmp-SLF to modulate gene expression in a yeast three-hybrid assay. Finally, we engineered the Golgi-resident glycosyltransferase FucT7 for tunable control by Tmp-SLF in mammalian cells.     

Improved interfacial properties of PI composites through graphene oxide and carbon nanotubes on carbon fiber surface

Carbon nanotubes (CNTs) have been identified as excellent nanoreinforcements for carbon fiber (CF)–reinforced polymers regarding a wide range of engineering applications. The outstanding properties of CNTs, such as their large surface area, high mechanical strength, and low manufacturing cost bring them to be distinguished nanoreinforcements for carbon fiber–reinforced polymers to form multifunctional and multiscale composites. Electrophoretic deposition of graphene oxide for CNTs onto the CF surface was conducted. The presence of graphene oxide–CNTs may effectively increase both the roughness and wettability of the CF surface, resulting in an improvement to the interfacial bonding strength between the CF and the polyimide (PI).     

"Fingertip"-guided noncovalent functionalization of carbon nanotubes by dendrons

Noncovalent functionalization of carbon nanotubes (CNTs) by dendrons was demonstrated. Certain types of dendrons successfully functionalized CNT surfaces through the noncovalent interactions between the peripheries of the dendrons and the sidewalls of CNTs. Dendrons have a unique anisotropic shape and an orthogonal functional group at their apex, and thus can generate a certain spacing between the functional groups upon immobilization on surfaces. Atomic force microscope (AFM) imaging, dispersion experiments, and MicroRaman spectroscopy were employed for the characterization of the functionalization. The binding was found to be governed by the chemical nature of the terminal groups, namely, the "fingertips", through a comparison study on the adsorption efficiency of the dendron analogs. Functional groups such as the carboxylic acid group and the benzyl amide group were effective for the cooperative binding. AFM analysis showed that the average spacing generated by the dendrons was 14-15 nm at a particular adsorption condition. Assembling streptavidin on the tubes through the dendrons and biotin confirmed the realization of the regulated spacing as well as the elimination of unwanted aggregation. The noncovalent functionalization of CNTs by a dendron can be a new approach toward sensible nanobiodevices, not only by introducing biomolecular probes on CNTs without disruption of the electronic network of the tubes, but also by providing the immobilized probe molecules with a space ample enough to minimize steric hindrance for the unhindered interaction with their target species.