基于核酸适体电化学生物传感器用于腺苷的免标记检测

以纳米MnO2作为适体固定的构建平台,制备了一种基于核酸适体的新型腺苷电化学生物传感器.固定于电极表面的适体探针与目标腺苷杂交后使电极界面的结构发生改变,通过[Fe(CN)6]3-/4-氧化还原探针监测传感器表面电子传递电阻的变化,以此作为检测信号进行腺苷的免标记检测.表面电子传递电阻的变化值与腺苷浓度的对数在1.0×...     

面向活体分析的核酸适体电化学生物传感研究

化学是生命活动的物质基础.在活体层次上精准获取生理活性分子的分布和含量的变化规律对于了解和研究生理和病理过程具有重要的意义.电化学方法,特别是基于核酸适体的电化学生物传感器,由于兼具高特异性识别、检测对象广以及易于微型化等优良特性,已成为复杂生理环境中实现快速、灵敏和高选择性检测的有效工具.本文综述了核酸适体电化学生物传感器的设计以及在活体分析中的应用,并对其未来发展趋势进行了展望.     

基于核酸适体的电化学生物传感器*

核酸适体是一类体外筛选的、可与目标分子高效、高特异亲合的RNA或DNA寡核苷酸片段,与常规识别分子（如抗体等）相比,核酸适体作为一类新型识别分子具有明显特色和优势,已被广泛应用于生物传感等分子识别和应用研究领域。本文就基于核酸适体的电化学生物传感器（标记型和非标记型）的近期进展作简要评述,包括适体简介、标记型（“信号衰减”型、“信号增强”型、酶标记型和纳米粒子标记型）和非标记型电化学适体生物传感器等内容。     

电化学和电化学发光核酸适体传感器

核酸适体具有亲合力强、选择性高、稳定性好、易于修饰等优点,广泛用于对目标物如蛋白质、小分子等的灵敏检测.电化学具有成本低、灵敏度高、仪器小巧等优点.近年来,构建基于核酸适体的电化学传感器,已经成为一个热门的研究领域.本文重点评述了2005年以来核酸适体的电化学传感器的研究进展,并展望其发展前景.     

基于核酸适体的电化学生物传感研究

核酸适体是一类经由指数富集的配体系统进化（SELEX）技术在体外筛选获得的单链寡核苷酸片段,由于具有可人工批量合成、价格低廉、易于功能化修饰、特异性强、亲和力高、免疫原性低、批间差异小、热稳定性好等优良特性,在分析化学、疾病治疗以及生物医学研究等诸多领域备受关注。结合代表性案例,该文综述了核酸适体在电化学生物传感领域的应用。首先简要概述了核酸适体及电化学适体传感器的特点,分类阐述了电化学适体传感器在小分子化合物、蛋白质、外泌体、循环肿瘤细胞（CTCs）以及病原微生物检测中的应用,并重点介绍了相关检测方法的原理、分析特性以及所应用的信号放大策略,最后对电化学适体传感器的发展进行了展望。     

基于核酸适体的试纸条检测三磷酸腺苷

将核酸适体（Aptamer）的特异性与纳米金颗粒的独特光学性质相结合,制备了一种适用于小分子检测的干式试纸条。该试纸条以修饰功能化核酸适体（PloyT13-Aptamer）的纳米金为识别元件,在控制线（C）上修饰ployA13序列,与识别元件结合以判断试纸条的有效性;在测试线（T）上修饰与Aptamer部分互补的DNA序列,与待测物形成竞争关系,通过T线上纳米金显色的深浅来定性或定量分析待测物浓度。结果表明,优化实验条件下,观察T线颜色变化可实现三磷酸腺苷（ATP）的肉眼定性检测,视觉检出限为10μmol/L。使用Image J软件进行定量分析,试纸条检测范围为10～1000μmol/L,检出限为2.4μmol/L。该试纸条生物传感器可以在10 min内得出检测结果且特异性良好,血清中回收率为104.4%～119.0%,为ATP的现场快速检测提供了一种经济有效的方法。     

基于核酸适体构象变化的荧光钾离子检测

噻唑橙(Thiazole orange,TO)在游离状态下几乎没有荧光,但当其与富G的DNA结合后,会产生很强的荧光增强效应。据此,利用TO识别有无钾离子存在时富G序列的构象变化,建立了一种基于核酸适体(Aptamer)构象效应进行钾离子检测的无标记荧光检测方法。在优化实验条件下,钾离子浓度在(1～60)×10-6mol/L和(4～9)×10-4mol/L范围内与荧光强度呈良好的线性关系,检出限(S/N=3)为2.2×10-7mol/L。对Li+、Na+、Ca2+、Mg2+、NH4+5种离子进行对照试验的结果表明,该方法不仅具有较好的灵敏度还具有较好的选择性,且无需对DNA进行标记,成本低,分析速度快。     

核酸适体生物传感器*

构建高速度、高特异性、高灵敏的蛋白质检测技术是目前蛋白质组学研究所面临的紧迫任务。传统蛋白质的检测主要利用抗体-抗原的特异相互作用。利用寡核苷酸间的严格的识别和亲和力而设计的人工合成寡核苷酸-适体（aptamer）的出现,使抗体抗原反应发生新的革命性变化。核酸适体对蛋白质的结合力和特异性可与抗原抗体间的作用力相媲美,且与抗体相比有许多优越性。因此利用核酸适体构建蛋白质的检测方法己引起许多科学工作者的关注。本文综述了核酸适体的发现(包括SELEX技术的原理),特点, 核酸适体生物传感器的原理、分类和应用,并对核酸适体生物传感器的发展进行了展望。     

无标记型核酸适体荧光传感器检测氯霉素的含量

建立了DNA核酸适体检测氯霉素的荧光分析新方法。将前人筛选得到的80bp的氯霉素DNA核酸适体截短至40bp,实验发现截短并不影响核酸适体与氯霉素的结合能力。40bp的DNA核酸适体与另一条互补DNA链形成双链DNA,SYBR Green I荧光染料能插入双链并有强荧光发射,加入氯霉素后,氯霉素能和DNA核酸适体形成稳定的复合物,诱导DNA双链打开,SYBR Green I释放,荧光降低。实验结果显示:在10~200μmol/L范围内,荧光降低百分数和氯霉素之间有良好线性关系,检测限为6μmol/L。     

一种胶质瘤细胞的核酸适体电化学阻抗传感器

本文基于交流阻抗谱技术发展了一种新型的高灵敏核酸适体传感器用于胶质瘤细胞的检测。该传感器通过巯基在金表面的混合自组装,将腱肽蛋白c的核酸适体探针固定于金电极表面。基于腱肽蛋白在胶质瘤细胞表面的高表达,利用细胞与电极表面核酸适体探针的特异性生物识别对铁氰化物电化学阻抗的抑制,建立了胶质瘤细胞检测的核酸适体传感器。考察并优化了核酸适体/巯基丙酸混合自组装比例、反应时间、温度和离子强度等对传感器性能有显著影响的分析条件,结果表明该传感器的阻抗响应与胶质瘤细胞浓度呈线性关系,线性范围为100~50 000 cell/mL,其检出限可达50 cell/mL。     

A Label-Free Fluorescence Aptasensor Based on G-Quadruplex/Thioflavin T Complex for the Detection of Trypsin

A novel, label-free fluorescent assay has been developed for the detection of trypsin by using thioflavin T as a fluorescent probe. A specific DNA aptamer can be combined by adding cytochrome c. Trypsin hydrolyzes the cytochrome c into small peptide fragments, exposing the G-quadruplex part of DNA aptamer, which has a high affinity for thioflavin T, which then enhances the fluorescence intensity. In the absence of trypsin, the fluorescence intensity was inhibited as the combination of cytochrome c and the DNA aptamer impeded thioflavin T’s binding. Thus, the fluorescent biosensor showed a linear relationship from 0.2 to 60 μg/mL with a detection limit of 0.2 μg/mL. Furthermore, the proposed method was also successfully employed for determining trypsin in biological samples. This method is simple, rapid, cheap, and selective and possesses great potential for the detection of trypsin in bioanalytical and biological samples and medical diagnoses.     

碳纳米管的功能化及其电化学性能

超级电容器作为一种新型的储能元件,以其快速储存、释放能量等优点,近年来成为各国科研工作的研究重点和焦点[1￣3],并在数据记忆存储系统、便携式仪器设备、后备电源、通讯设备、计算机、燃料电池、电动车混合电源等许多领域都有广泛的应用前景[4]。目前,超级电容器用的电极材     

Electrochemical Biosensing Systems Based on Carbon Nanotubes and Carbon Nanofibers

Abstract

Carbon nanomaterials are in the forefront of research in a variety of chemical and physical disciplines. Of these, certain nanostructures seem to be suitable for the development of electrochemical biosensors. In particular carbon nanotubes, and carbon nanofibers have specific chemical and physical characteristics that lent them ideal for the development of biosensors with unique analytical characteristics. In particular, their conductivity, surface area, inherent and induced chemical functionalities, and biocompatibility provide the grounds for the development of a new era of electrochemical biosensors. In this review, we will examine the recent developments of biosensor design based on these new nanostructures.     

多壁碳纳米管结构与其电化学容量之间关系的研究

采用化学气相沉积法,通过改变催化剂的成分、碳源、反应和后处理条件来制 备不同管径、管长、石墨化程度的多壁碳纳米管。经电化学容量性能测试、透射电 子显微镜观察和N_2吸附等结构表征,发现管径分布为30.0～40.0 nm、管长越短、 石墨化程度越低、比表面积越大、孔容越大的多壁碳纳米管具有更好的电化学容量 。     

A Label‐Free Electrochemical Aptasensor for Thrombin Using a Single‐Wall Carbon Nanotube (SWCNT) Casted Glassy Carbon Electrode (GCE)

An electrochemical aptasensor with a thrombin binding aptamer (TBA) was developed using a single‐wall carbon nanotube (SWCNT) casted GCE. The TBA was immobilized on SWCNTs through π‐stacking without any special modification, resulting in helical wrapping to the surface. In the presence of thrombin, the TBA binds with thrombin and the TBA concentration on the SWCNT surface decreases. The remaining amount of TBA can be analyzed by an electrochemical method without any label, because the guanine bases of the nucleic acid are measurable by electrochemical methods. The electrochemical oxidation of guanine nucleotides was enhanced by electrocatalytic mediation using Ru(bpy)₃²⁺ for higher sensitivity and reduction of the overpotential for electrochemical detection.     

基于 G-四联体门控制效应的铅离子适配体传感器

将富鸟嘌呤(G)序列核酸适配体与门控制效应相结合,通过控制门的开关实现信号放大,构建了新型电化学生物传感器,用于铅离子(Pb2+)的高灵敏检测。首先将单-6-巯基-β-环糊精(6-SH-β-CD)自组装在金电极上,形成有序排列的分子自组装膜,且分子之间留有空隙,可作为探针通过的门结构。随后将发夹结构的富含 G 序列的核酸适配体修饰在环糊精次面端口,制得可特异性识别 Pb2+的电化学生物传感器。富 G 序列适配体结合 Pb2+后可折叠形成 G-四联体结构(G4-Pb2+),覆盖住电极表面的探针通道,产生关门效应,使探针氧化还原电流强度减小,进而形成门控制效应,利用该效应可进行 Pb2+的定量检测。门控制效应显著提高了信噪比和检测的灵敏度,在1×10-13~5×10-11 mol/ L 浓度范围内,Pb2+浓度的负对数与 DPV 响应电流呈良好的线性关系,检出限为3.6×10-14 mol/ L(DL=3δb / K)。传感器用于实际水样品中 Pb2+的测定,结果令人满意。     

Electrodeposition of Cobalt Oxides on Carbon Nanotubes for Sensitive Bromhexine Sensing

We develop an electrochemical sensor for the determination of bromhexine hydrochloride (BHC), a widely use mucolytic drug. The sensor is prepared by electrodeposition of cobalt oxides (CoO_x) on a glassy carbon electrode modified with carboxylated single-walled carbon nanotubes (SWCNT). A synergistic effect between CoO_x and SWCNT is observed, leading to a significant improvement in the BHC electrooxidation current. Based on cyclic voltammetry studies at varying scan rates, we conclude that the electrochemical oxidation of BHC is under mixed diffusion–adsorption control. The proposed sensor allows the amperometric determination of BHC in a linear range of 10–500 µM with a low applied voltage of 0.75 V. The designed sensor provides reproducible measurements, is not affected by common interfering substances, and shows excellent performance for the analysis of BHC in pharmaceutical preparations.     

Applications of Carbon Nanotubes in Electrochemical DNA Biosensors

The discovery of carbon nanotubes (CNTs) about a decade ago has brought fascinating evolutions in electronics, material industry, as well as bio-techniques for DNA analysis, gene therapy, drug delivery etc. It has also dramatically promoted the development of DNA biosensing techniques, especially electrochemical DNA biosensor. The application of CNTs in electrochemical DNA biosensors includes two main aspects: on one hand, using CNTs as a novel substrate not only enables immobilization of DNA molecules but also serves as a powerful amplifier to amplify signal transduction event of DNA hybridization. On the other hand, CNTs can also be employed as a powerful carrier to pre-concentrate enzymes or electroactive molecules for electrochemical sensing of DNA hybridization as a novel indicator. In this review, we place emphasis on recent studies of CNTs-based electrochemical DNA biosensors based on these two aspects, with advantages and disadvantages of each aspect introduced herein.     

一种估算多壁碳纳米管电化学容量的方法

Five kinds of multi-walled carbon nanotubes were prepared by CVD(Chemical Vapour Deposition) and purified under the same conditions, then were used as electrodes for super capacitors in order to compare their performance. The differences measured in terms of specific capacitance by galvanostatic cycling were presented, porosity measurements using nitrogen at 77 K made on carbon nanotubes allowd a better understanding of the electrochemical behavior of these carbon nanotubes. The experiment results show that pore diameter in the range of more than 3nm is required to maximize the capacitance in the electrolyte of 1.0 mol·L^-1 LiClO₄/EC+DEC(V_EC∶V_DEC=1∶1), and the BET surface area contributed by this extent is almost linear with the specific capacitance, in which an average value is about 0.11 F·m^-2, and the specific capacitance estimated from above is very close to the value by experiment.