多层生物介质传感器的特性研究（英文）

目的 多层生物介质的生物传感器被广泛应用于各大领域,其检测特性对于传感器优劣的评估尤为重要。本文目的在于量化表征多层生物介质的电学特征。方法 基于生物电阻抗谱技术来探究多层生物介质的电化学阻抗谱特性,并结合保角映射的方法来量化表征多层生物介质,阐明其对阻抗的影响规律,继而为生物传感器的研制与开发提供理论基础。有效提取各生物介质层修饰后电阻抗参数(Z^*),从而量化表征多层生物介质层的电阻抗谱特性。结果 对多层模型进行了理论计算并构建了相关试验测试系统,研究结果表明,随着生物介质层的逐步修饰,检测区域电阻抗参数(Z^*)在f=0.1～50 MHz下持续上升,理论计算结果趋势与试验结果趋势较好吻合,论证了此理论计算方法的正确性。结论 本文证实了可根据生物电阻抗谱和保角映射方法量化表征多层生物介质的电阻抗谱特性,对生物传感器的研制与开发有一定的实用价值。     

细菌内毒素电化学生物传感器的构建及性能表征

细菌内毒素为外源性致热源,内毒素的检测在生物制品生产过程中至关重要。构建了一种用于检测细菌内毒素的电化学核酸适体生物传感器,以氨基修饰的内毒素核酸适体EAQ2为配体,通过3-巯基丙酸(MPA)中间连接物,固定修饰在金电极表面,并通过循环伏安(CV)和电化学交流阻抗谱(EIS)两种方式共同表征了生物传感器的构建过程。结果发现MPA组装时间在6 h时能在金电极表面形成稳定的自组装单分子层。构建的生物传感器检测限达0.001 EU/ml,低于目前报道的其他内毒素检测方法的检测限,在0.001~0.1 EU/ml内毒素浓度范围内具有较好的线性关系,相关系数R~2=0.9878,在实际生物样品的检测上具有一定的应用前景。     

微阵列电化学生物传感器研究进展

本文简要介绍了微阵列电化学生物传感器的基本原理和分类,评述了微阵列电化学生物传感器的研究进展。     

阵列电化学生物传感器研究进展

阵列生物传感器技术作为一种高通量、快速、选择性高和集成化的分析技术,已在基因组学和蛋白质组学的研究和药物筛选、环境分析,食品分析,临床诊断等领域中得到广泛的应用.阵列生物传感器主要有阵列光学生物传感器和阵列电化学生物传感器.阵列电化学生物传感器是将生物分子识别物质如酶、抗原/抗体、DNA等固定在阵列电极上,以阵列中每根电极产生的电化学信号作为检测信号的电化学分析器件.阵列电化学生物传感器以灵敏度高、分析速度快、选择性好、易于微型化和集成化以及仪器价格低廉等特点受到了研究工作者的极大关注.本文简单介绍了阵列电化学生物传感器的原理和特点,重点评述了2005年以来阵列电化学生物传感器在单组份检测和多组份同时检测两方面的研究进展,简单讨论了阵列电化学生物传感器研究中存在的问题.     

DNA生物传感器研究进展

本根据作用机理不同将ＤＮＡ生物传感器分为ＤＮＡ光化学传感器,ＤＮＡ电化学传感器和压电晶体传感器,并就几种方面的研究进展进行了综述。     

纳米多孔硅阻抗生物传感器的研究

构建了1种基于多孔硅材料,无需标记的纳米生物传感器,用于对牛血清白蛋白分子进行检测。通过对多孔硅进行表面处理,形成氧化膜,将抗体固定到多孔硅氧化层表面。在磷酸盐缓冲液中,通过电化学检测系统检测加入抗原后,传感器的阻抗值的变化。磷酸缓冲液（PBS）/抗体-氧化层/硅,构成电解液/绝缘层/半导体（electro-lyte-insulator-semiconductor,EIS）结构。传感器的线性检测范围为0.01～0.27mg/mL,检测限为0.01mg/mL。     

DNA生物传感器及其研究进展

就ＤＮＡ生物传感器的工作原理,分类、ＤＮＡ探针的固化方法,以及电化学ＤＮＡ生物传感器、光学ＤＮＡ生物传感器及压电ＤＮＡ生物传感器的研究进展、优缺点和发展趋势加以介绍。     

阵列生物传感器研究进展

简要介绍了阵列生物传感器的基本原理和分类。根据换能器的不同,评述了光学、电化学、质量型、磁致阻抗等阵列生物传感器的研究进展,并对目前阵列生物传感器研究中存在的问题进行了分析。     

阵列生物传感器研究进展

简要介绍了阵列生物传感器的基本原理和分类。根据换能器的不同,评述了光学、电化学、质量型、磁致阻抗等阵列生物传感器的研究进展,并对目前阵列生物传感器研究中存在的问题进行了分析。     

电化学生物传感器快速检测DNA研究进展

本简要地介绍了DNA电化学生物传感器研究的最新进展,重点讨论了改善生物传感器选择性和灵敏度的技术和方法。     

Electrochemical impedance spectroscopy for study of aptamer-thrombin interfacial interactions

A simple and highly sensitive electrochemical impedance spectroscopy (EIS) biosensor based on a thrombin-binding aptamer as molecular recognition element was developed for the determination of thrombin. The signal enhancement was achieved by using gold nanoparticles (GNPs), which was electrodeposited onto a glassy carbon electrode (GCE), as a platform for the immobilization of the thiolated aptamer. In the measurement of thrombin, the change in interfacial electron transfer resistance of the biosensor using a redox couple of [Fe(CN)₆]^3−/4− as the probe was monitored. The increase of the electron transfer resistance of the biosensor is linear with the concentration of thrombin in the range from 0.12 nM to 30 nM. The association and dissociation rate constants of the immobilized aptamer–thrombin complex were 6.7 × 10³ M⁻¹ s⁻¹ and 1.0 × 10⁻⁴ s⁻¹, respectively. The association and dissociation constants of three different immobilized aptamers binding with thrombin were measured and the difference of the dissociation constants obtained was discussed. This work demonstrates that GNPs electrodeposited on GCE used as a platform for the immobilization of the thiolated aptamer can improve the sensitivity of an EIS biosensor for the determination of protein. This work also demonstrates that EIS method is an efficient method for the determination of association and dissociation constants on GNPs modified GCE.     

A Microfluidic-based Electrochemical Biochip for Label-free DNA Hybridization Analysis

Miniaturization of analytical benchtop procedures into the micro-scale provides significant advantages in regards to reaction time, cost, and integration of pre-processing steps. Utilizing these devices towards the analysis of DNA hybridization events is important because it offers a technology for real time assessment of biomarkers at the point-of-care for various diseases. However, when the device footprint decreases the dominance of various physical phenomena increases. These phenomena influence the fabrication precision and operation reliability of the device. Therefore, there is a great need to accurately fabricate and operate these devices in a reproducible manner in order to improve the overall performance. Here, we describe the protocols and the methods used for the fabrication and the operation of a microfluidic-based electrochemical biochip for accurate analysis of DNA hybridization events. The biochip is composed of two parts: a microfluidic chip with three parallel micro-channels made of polydimethylsiloxane (PDMS), and a 3 x 3 arrayed electrochemical micro-chip. The DNA hybridization events are detected using electrochemical impedance spectroscopy (EIS) analysis. The EIS analysis enables monitoring variations of the properties of the electrochemical system that are dominant at these length scales. With the ability to monitor changes of both charge transfer and diffusional resistance with the biosensor, we demonstrate the selectivity to complementary ssDNA targets, a calculated detection limit of 3.8 nM, and a 13% cross-reactivity with other non-complementary ssDNA following 20 min of incubation. This methodology can improve the performance of miniaturized devices by elucidating on the behavior of diffusion at the micro-scale regime and by enabling the study of DNA hybridization events.     

黄曲霉毒素氧化酶/壳聚糖-单壁碳纳米管/聚邻苯二胺修饰电极对杂色曲霉素的检测

将黄曲霉毒素氧化酶(AFO)固定在壳聚糖(CS)-单壁碳纳米管(SWCNTs)杂交膜中,组装在聚邻苯二胺(POPD)修饰的金电极(Au)表面,制备了对杂色曲霉素(ST)敏感的生物传感器(AFO/CS-SWCNTs/POPD/Au)。运用原子力显微镜(AFM)、傅立叶变换红外光谱(FT-IR)和交流阻抗技术(EIS)对电极组装过程进行了表征。循环伏安法研究表明,AFO在修饰电极上发生了准可逆的氧化还原反应,是表面控制过程,其式量电位为-0.436V(vs.Ag/AgCl),说明包埋在CS-SWCNTs中的AFO和电极之间发生了直接电子传递。AFO修饰电极对ST具有明显的电催化作用,其表观米氏常数appKM为7.13μmol/L,催化电流与ST浓度在10～310ng/mL范围内呈线性关系,相关系数为0.997,检出限为3ng/mL(S/N=3),响应时间小于10s。组装的生物传感器具有较好的稳定性与重现性,连续检测20ng/mL的ST标准溶液11次,电流值RSD为3.9%;放置一个月后,其电流响应值仍为初始值的85.6%。该方法具有较高的选择性和灵敏度,应用于实际样品检测时,其回收率在87.6%～105.5%之间...     

Development of a new kind of dual modulated QCM biosensor

To distinguish the mass loading effect from the total frequency change is a problem in the application of a quartz crystal microbalance (QCM) biosensor in the liquid phase. Based on the characteristic damping theory, this paper proposes a new method of dual modulation to solve this problem. Usingg polyethyleneimine to immobilize anti-SE (staphylococcin enterotoxin) antibody (C₂ type), a dual modulated QCM SE biosensor was developed and the experiment proved that it has little cross-reaction with B-type SE. The measuring curve of the sensor was also determined through experiment.     

Impedance studies of a nano-structured conducting polymer and its application to the design of reliable scaffolds for impedimetric biosensors

Electrochemical impedance spectroscopy (EIS) as a powerful, non-invasive and informative technique was used to obtain important information about kinetics of doping process in conducting polymers. Polypyrrole (PPy) and its derivatives can form conducting polymer films which represent excellent charge transfer behaviors during doping processes. It can also have a wide range of applications in bioelectrochemistry. In the present study the conducting polymer of alpha-carboxy pyrrole (alpha-COOH-PPy), appended onto the underlying film of PPy, was prepared by electrochemical methods and its behavior was analyzed using EIS. From highly accurate fitting of impedance results it was found that the charging mechanism is governed by the diffusion process. In addition, the impedance analyses provided values for the bulk polymer parameters including diffusion coefficient (D), equilibrium capacitance (C(0)) and diffusion resistance (R(0)). The surface morphology of the polymeric film was characterized using scanning electron microscopy (SEM). The film was then used to immobilize the cytochrome C (cyt-C) and to perform its electrochemical studies. The modified cyt-C/alpha-COOH-PPy electrode was used for electrocatalytic reduction of H(2)O(2) in solution and its viability as a new impedimetric biosensor was examined. Based on the calibration curve obtained for the proposed impedimetric biosensor, the limit of detection and relative standard deviation were evaluated as 0.25mumolL(-1) and 7%, respectively. Finally, the prolonged stability test was performed and high stability and reproducibility of the new biosensor was confirmed.     

Engineered allosteric ribozymes as biosensor components

RNA and DNA molecules can be engineered to function as molecular switches that trigger catalytic events when a specific target molecule becomes bound. Recent studies on the underlying biochemical properties of these constructs indicate that a significant untapped potential exists for the practical application of allosteric nucleic acids. Engineered molecular switches can be used to report the presence of specific analytes in complex mixtures, making possible the creation of new types of biosensor devices and genetic control elements.     

Construction of an amperometric glucose biosensor based on the immobilization of glucose oxidase onto electrodeposited Pt nanoparticles-chitosan composite film

One-step construction of Pt nanoparticles-chitosan composite film (PtNPs-CS) was firstly proposed as a novel immobilization matrix for the enzymes to fabricate glucose biosensor. This novel interface embedded in situ PtNPs in CS hydrogel was developed by one-step electrochemical deposition in solution containing CS and chloroplatinic acid (H(2)PtCl(6)). Several techniques, including scanning electron microscopy (SEM), cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and chronoamperometry were employed to characterize the assembly process and performance of the biosensor. Under the optimized experimental conditions, the resulting biosensor exhibited excellent linear behavior in the concentration range from 1.2 μM to 4.0 mM for the quantitative analysis of glucose with a limit of detection of 0.4 μM at a signal-to-noise ratio of 3. The apparent Michaelis-Menten constant (K(M)(app)) was evaluated to be 2.4 mM, showing good affinity. The proposed biosensor offered good amperometric responses to glucose due to the nanostructured sensing film provided plenty of active sites for the immobilization of glucose oxidase (GOD).     

Development of DNA electrochemical biosensor based on immobilization of ssDNA on the surface of nickel oxide nanoparticles modified glassy carbon electrode

A sensitive electrochemical method for DNA hybridization based on immobilization of DNA probe and [Ru(NH₃)₅Cl]PF₆ complex onto nickel oxide nanomaterials (NiOx_np) modified glassy carbon electrode was developed. Due to strong affinity of NiOx_np for phosphate groups, oligonucleotides probe with a terminal 5′-phosphate group was attached to the surface of the modified electrode. DNA immobilization and hybridization were characterized by electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry using K₃Fe(CN)₆/K₄Fe(CN)₆ and [Ru(NH₃)₅Cl]PF₆ as probe and indicator, respectively. The Ru-complex current response indicates only the complementary sequence showing an obvious current signal in comparison to non-complementary and three or single point mismatched sequences. The fabricated biosensor possessed good selectivity and sensitivity for complementary probe, taxon: 32630 tumor necrosis factor (TNF). The linear dynamic range, sensitivity and detection limit of the proposed biosensor were 4 × 10⁻¹⁰ M to 1 × 10⁻⁸ M, 34.32 nA nM⁻¹ and 6.8 × 10⁻¹¹ M, respectively. Excellent reproducibility and stability, quite simple and inexpensive preparation are the other advantages of proposed biosensor.     

Electrochemical impedance detection of DNA hybridization based on dendrimer modified electrode

Bioactive ultrathin films with the incorporation of amino-terminated G4 PAMAM dendrimers have been prepared via layer-by-layer self-assembly methods on a gold electrode and used for the DNA hybridization analysis. Surface plasmon resonance (SPR), X-ray photoelectron spectroscopy (XPS), and electrochemical impedance spectroscopy (EIS) are used to characterize the successful construction of the multicomponent film on the gold substrate. The dendrimer-modified surfaces improve the immobilization capacity of the probe DNA greatly, compared to the AET (2-aminoethanethiol) SAM sensor surfaces without dendrimer molecules. DNA hybridization analysis is monitored by EIS. The dendrimer-based electrochemical impedance DNA biosensor shows high sensitivity and selectivity for DNA hybridization assay. The multicomponent films also display a high stability during repeated regeneration and hybridization cycles.