基于纳米金标记和银增强放大的血吸虫抗体的免疫分析

该文提出了一种新的、高灵敏的基于银增强放大和免疫胶体金技术的血吸虫抗体的免疫分析方法.免疫反应在聚苯乙烯微孔板中以间接分析模式(血吸虫抗原-兔抗血吸虫抗体-纳米金标记的羊抗兔IgG二抗)进行,通过反应上去的纳米金催化银离子的还原,使大量的银沉积在纳米金的周围,用酸将银溶解后,通过阳极溶出伏安法(ASV)对银离子进行检测,溶出峰电量的大小与待分析物血吸虫抗体(一抗)的浓度成正比.对免疫分析的一些实验条件如抗原与待测抗体的反应时间、纳米金标记二抗与待测抗体的反应时间、纳米金标记抗体稀释度以及银增强时间进行了优化.阳极溶出峰电量与日本血吸虫抗体的浓度在6.4μg/L～100mg/L范围内呈良好的线性关系,检测下限为3.0 μg/L.将该方法应用于兔血清中日本血吸虫抗体的测定,取得了满意的结果.     

结合PAH聚电解质和纳米金界面的压电免疫传感器研究

提出了一种聚丙烯氯化铵(PAH)-纳米金固定抗体的压电免疫传感器界面的构建方法.先在压电石英晶振的金电极表面自组装一层半胱氨酸单层膜,通过戊二醛交联带大量NH2基的聚电解质PAH,随后在PAH膜表面自组装一层纳米金粒子,以静电吸附作用固定IgG抗体,研制成一种新的压电免疫传感器的界面,用于对相应抗原的检测.研究了PAH浓度及抗体固定化等实验条件的影响,探讨了传感器的主要响应特性与再生性能,并与戊二醛直接固定的传感器的性能进行了比较.结果表明,前者固定的抗体的活性较高,响应频率较大,检测的线性范围较宽,非特异性吸附小,能有效地改善传感器的灵敏度和检测限,而且容易进行传感器的再生.     

压电免疫传感器

本文概要介绍了压电免疫传感器的原理和生物活性物质的固定化方法，并对文献中的各类压电免疫传感器的发展作了综述。     

压电免疫传感器的实现

压电免疫传感器是利用压电石英晶体对表面电极区附着质量的敏感性,并依据生物功能分子（如抗原和抗体）之间的选择特异性,使压电晶体表面产生微小的压力变化,引起其振动频率改变。在此原理基础上可实现对多种抗原或抗体进行实肘、快速地定量测定,具有高特异性、高灵敏度、响应快和小型简便等特点。     

压电晶体式涡街流量传感器放大电路的设计

通过对涡街信号频谱的分析，发现涡街传感器主要干扰源为固有振荡频率，并设计了解决此种干扰的传感器放大电路。     

基于纳米金/硫堇修饰金电极的脱落酸安培免疫传感器的研制

提出了一种基于纳米金/硫堇修饰金电极的ABA安培免疫传感器。该传感器基于H2O2-HRP-硫堇催化波体系构建,其中硫堇为传感介质。当HRP存在时,通过加入H2O2,硫堇的还原电流大幅增加,并且电流的增加依赖于HRP活性。HRP活性又由ABA与HRP酶标抗体结合物调控,产生一个减小的催化波。用BSA封闭硫堇单分子层修饰后可能存在的活性位点以避免非特异性吸附。优化了测定条件,包括酶标抗体和硫堇的最佳比例、培育时间、缓冲液的pH值和H2O2浓度。此传感器的还原电流在ABA浓度0.5~1000ng/mL范围内呈线性下降,回归方程为y=0.0209x 17.071,相关系数为0.9922,检测限为0.2ng/mL。     

压电免疫传感器的研制及应用

介绍了压电免疫传感器的压电质量传感原理、免疫反应及压电免疫传感过程。同时将该传感器成功地应用于牛血滑白蛋白的检测．     

自组装纳米金内毒素压电传感器的研究

利用纳米金颗粒自组装于金片表面上,用于固定多粘菌素B(PMB),研制了一种内毒素压电传感器。先在石英晶体电极表面自组装1,6—己二硫醇,再通过纳米金与巯基的共价键合作用,在巯基自组装膜的另一端的巯基表面形成一均匀的纳米金单层膜,实现PMB在传感器表面的固定。实验探讨了影响纳米金自组装和PMB固定等主要实验参数和条件;考核了采用此固定化方法传感器的响应性能。结果表明:与单纯采用自组装方法比,传感器在灵敏度和分辨力方面均有较大的改善。     

压电免疫传感器用于转铁蛋白免疫反应的动力学研究

采用蛋白 A 将抗体固定在9MHz 双面镀金石英晶体上,研制了一种用于转铁蛋白检测的压电免疫传感器,并用该技术监测了转铁蛋白抗原在压电石英晶体上的吸附与解吸过程。研究了蛋白 A 浓度、抗体效价及抗原抗体反应时间等对传感器灵敏度的影响。着重研究了免疫反应体系的反应动力学。结果表明:转铁蛋白在压电石英晶体上的键合行为符合 Langmuir 吸附方程,免疫反应的结合常数 K_(ass)为6.75×10~7(mol/L)~(-1),并计算了该免疫反应的速率常数 K_f 为2.14×10~7(mol/L)~(-1)S~(-1)。     

聚邻氨基苯甲酸/纳米金复合膜电化学免疫传感器用于水体中大肠杆菌的检测

以聚邻氨基苯甲酸/纳米金复合膜(poly-o-ABA/AuNP)为基底固载大肠杆菌抗体制备了一种新型的电化学免疫传感器.利用大肠杆菌与抗体之间的免疫反应构建了三明治夹心结构,以对苯二酚为电子媒介体,通过辣根过氧化酶(HRP)催化H2O2产生的响应电流对大肠杆菌进行检测.结果表明,在优化的实验条件下,该传感器的响应电流与大肠杆菌的浓度在1.0×103～1.0×107cfu/mL范围内呈良好的线性关系,检测限为2×102cfu/mL.经过预富集过程,该方法成功实现了对河水中大肠杆菌的检测.     

Large-area Raman enhancement substrates using spontaneous dewetting of gold films and silver nanoparticles deposition

We present a cost-effective method for making large-area surface enhanced Raman scattering (SERS) substrates by using spontaneous dewetting of ultrathin gold film. The dewetting of 5-nm-thick gold film formed high-density gold islands ranged from 40 nm to 80 nm. The measured SERS signal was 5 times stronger than synthesised gold nanoparticles. The SERS signals can be further increased by depositing small silver nanoparticles on the dewetted film. The experimental results showed 5-nm-thick silver coating increased SERS signals up to 10 times. The calculations by finite-difference time-domain method verified such SERS enhancement originated from enhanced electric fields between gold islands and silver nanoparticles. In the application, we demonstrated an all-optical measurement of pH values in microfluidic devices by using SERS signals of para-mercaptobenzoic acid.     

Performance enhancement of a piezoelectric linear array transducer by half-concave geometric design

A geometric design for array elements of a piezoelectric linear array transducer is proposed and evaluated. The design concept is based on the half-concave geometry, in which the radiating surface is concave while the other surface remains plane. A ∼1.8 MHz piezoelectric linear array transducer with half-concave elements has been designed, fabricated and evaluated. A dicing method was developed to shape the concave surfaces of the piezo elements and matching layers. By comparing the transducer performance, a traditional linear array transducer with similar dimension has been fabricated. It was found that the half-concave array transducer has significantly broader −6 dB bandwidth (96%), higher effective electromechanical coupling coefficient (0.62), and lower insertion loss (−21 dB) compared to those (76%, 0.55, and −25 dB, respectively) of the plane array transducer. The enhanced coupling coefficient and bandwidth are caused by the broaden resonance of the elements, which is induced by the continuously varying thickness in the designed geometry. The increased sensitivity is mainly attributed to the focused radiating surface.     

Characteristics of a label-free piezoelectric immunosensor detecting Pseudomonas aeruginosa

A label-free immunosensor system detecting a psychrophylic bacterium, Pseudomonas aeruginosa was developed as follows. Four types of anti-P. aeruginosa antibody were individually chemisorbed onto one-side gold electrodes of piezoelectric quartz crystals according to a thiolated antibody coupling procedure initiated with a thiol-cleavable heterobifunctional cross-linker, sulfosuccinimidyl-6-[3-(2-pyridyldithio)propionamido]hexanoate. A flow-type biosensor system was operated optimally at 0.2 M sodium potassium phosphate, pH 7.2 with a minimal matrix effect and the selected flow rate for it was 0.155 ml/min. A biosensor response was detected by measuring a steady-state resonant frequency shift after the response time around 8 min. The frequency shifts obtained were quite specific according to the antibody types and P. aeruginosa strains. The biosensor responses to varying concentrations of the P. aeruginosa cells ranging from 1.3×10⁷ to 1.3×10⁸ CFU/ml were determined as 17–176 Hz and a linear calibration curve (r=0.942) was obtained by plotting the responses in a double-logarithmic scale. The selectivity of the biosensor between P. aeruginosa and Xanthomonas spp. which both belong to the aerobic pseudomonads was, however, not so good owing to the property of the antibody used. The sensor chip could be reused at least seven times without an appreciable decrease in sensitivity.     

Conductimetric immunosensor for atrazine detection based on antibodies labelled with gold nanoparticles

A novel conductimetric immunosensor for atrazine detection has been designed and developed. This immunosensor is mainly based on antibodies labelled with gold nanoparticles. Additionally, the immunosensor consists of an array of two coplanar non-passivated interdigitated metallic μ-electrodes (IDμE) and immunoreagents specifically developed to detect this pesticide. The chemical recognition layer was covalent immobilized on the interdigital space. Immunochemical detection of the concentration of atrazine is achieved by a competitive reaction that occurs before the inclusion of the labelled antibodies. It is shown that the gold nanoparticles provide an amplification of the conductive signal and hence makes possible to detect atrazine by means of simple DC measurements.The conductimetric immunosensor and its biofunctionalization steps have been characterized by chemical affinity methods and impedance spectroscopy.This work describes the immunosensor structure, fabrication, physico-chemical and analytical characterization, and the immunosensor response using conductivity measurements. The immunosensor developed detects atrazine with limits of detection in the order of 0.1–1 μg L⁻¹, far below the maximum residue level (MRL) (100 μg L⁻¹) established by European Union (EU) for residues of this herbicide in the wine.Although in this paper the competitive reaction occurs in buffer, an initial study of the wine matrix effect is also described.     

Colorimetric immunoassay chip based on gold nanoparticles and gold enhancement

A colorimetric immunoassay chip has been developed based on gold nanoparticles for indicating the antibody–antigen binding activity and gold enhancement for amplifying the specific binding signal. Our investigations showed that the results of immunoassay can be represented by the level of color intensity. They were easily observed by a regular camera or naked eye, which is not needed of sophisticated laboratory equipment. Optimization of experimental conditions was carried out and the colorimetric detection had been compared to the standard chemifluorescent detection. Under the optimized conditions, colorimetric immunoassay chip had been demonstrated to detect different amount of immobilized antigens, i.e., human IgG. The results, i.e., color intensity, were mapped to the concentration of immobilized antigens in a dynamic range of 1–5,000 ng/ml. The proposed detection method does not require any sophisticated optical systems; therefore, it is possible to be miniaturized and integrated into a microfluidic system for developing a portable immunoassay device.     

Design and analysis of a novel piezoelectric inertial actuator with large stepping displacement amplified by compliant mechanism

Microsystem Technologies - A novel piezoelectric inertial actuator is developed in this paper, to realize the requirements of large stroke and high output displacement accuracy. This proposed...     

带质量块微型双稳态压电板的动力学分析

本文对带质量块的微型双稳态压电板进行动力学分析.以中心固支四边自由的带质量块微型压电层合板为研究对象,应用应变梯度理论考虑尺寸效应,综合考虑力、电、热耦合作用,采用Von Karman大变形理论,运用Hamilton原理建立非线性动力学方程.利用特征值法探究不同内禀长度和不同压电铺设面积的情况下,温度和电压对其固有频率和稳定性的影响.其次研究了不同外激励下系统的非线性动力学响应.通过本文的研究发现,随着压电铺设面积的增大,力、电、热耦合效应增强,对系统的稳定性影响越显著;通过研究温度和电压对系统振动幅值的影响为振动控制提供了理论依据.同时发现尺寸效应对结构刚度影响较大,验证了微型结构考虑尺度效应的必要性.本文的研究结果会为今后的工程实际应用提供一定的理论参考价值.     

Modeling and optimal design of piezoelectric cantilevermicroactuators

A novel model is described for predicting the static behavior of a piezoelectric cantilever actuator with an arbitrary configuration of elastic and piezoelectric layers. The model is compared to deflection measurements obtained from 500-μm-long ZnO cantilever actuators fabricated by surface micromachining. Modeled and experimental results demonstrate the utility of the model for optimizing device design. A discussion of design considerations and optimization of device performance is presented     

Compensator design for stability enhancement with collocatedcontrollers

A continuum model rather than a finite element model is used. The optimal compensator design is formulated as a stochastic regulator problem and is shown to be solvable by the general infinite-dimensional theory developed by the author despite the lack of exponential stabilizability. Infinite-dimensional steady-state Riccati equations characterizing the feedback control gain and the Kalman filter gain operators can be solved explicitly. The associated performance indexes including the mean-square control-effort are calculated in closed form. As a first approximation, the compensator transfer function can be realized as a bank of bandpass filters in parallel centered at the undamped mode frequencies. Numerical calculations for the gain and bandwidths for a typical configuration are presented. The performance of the compensator is evaluated when in fact in the true model there is no actuator noise. The theoretical problem involved is to show that the infinite-dimensional stochastic process is asymptotically stationary. It is possible to calculate the steady-state covariance in closed form and thereby calculate performance indexes of interest explicitly, facilitating the choice of optimal design parameters     

基于银染色放大和免疫胶体金的金属免疫分析

提出了一种新的、高灵敏的基于银染色放大和免疫胶体金技术的金属免疫分析方法.免疫反应在聚苯乙烯微孔板中以夹心分析模式进行,通过纳米金对银离子的催化还原在金标抗体上沉积大量的金属银,用酸溶解后,在玻碳电极上用阳极溶出伏安法(ASV)对银离子进行检测,溶出峰电流的大小间接与待分析物人免疫球蛋白G(IgG)的浓度成正比.对阳极溶出分析的测定参数如沉积电位、富集时间和免疫分析的一些条件如抗原-抗体反应时间、金标抗体稀释度等进行了优化.阳极溶出峰电流与IgG浓度在1.66 μg/L～27.25 mg/L范围内呈良好的线性关系,检测下限为1.1 μg/L.