SPR生物传感器及其应用进展

基于表面等离子体共振 (SPR)技术的光学生物传感器是进行生物分子相互作用分析的一种先进手段。与传统的超速离心、荧光法等相比 ,它具有实时检测、无需标记、耗样最少等特点 ,在药物筛选、临床诊断、食物及环境监控和膜生物学等领域中的新兴应用日益扩大 ,并且已成为生命科学和制药研究的一种标准的生物物理学工具。综述了近几年国际上生物传感器的应用进展情况 ,并简要展望了该技术的发展和应用前景     

一种葡聚糖芯片的再生方法、表征及其应用*

表面等离子体共振（surface plasmon resonance, SPR）生物传感器,作为一种适时快捷,无需标记的生物分子相互作用研究工具,已广泛应用于生物化学分析与研究。羧甲基化葡聚糖修饰的CM5传感芯片是Biacore 系列仪器应用最为普遍的核心部件,目前CM5芯片主要从法玛西亚公司购买,价格昂贵,且一旦共价交联的受体分子失活,就不能重复利用。阐述了一种简便、低成本、用于SPR生物传感器的葡聚糖修饰金膜芯片的再生方法及其表征和应用。用此方法再生的芯片能被循环伏安法和原子力显微镜很好地表征,并成功地用于抗前列腺特异性抗原(prostate-specific antigen,PSA)固定和PSA检测, 同时测定了PSA与其抗体之间的动力学和亲和常数。     

生物传感芯片质谱及其在蛋白质组研究中的应用

目前蛋白质组研究技术在已有的研究基础上取得了长足进展 ,包括固相 ｐＨ梯度双向凝胶电泳、生物质谱技术、蛋白质双向电泳图谱的数字化和各种分析技术、蛋白质间相互作用分析的方法如酵母双杂交系统、噬菌体展示、表面等离子共振等。但均各有其利弊。目前质谱是进行蛋白质组成分鉴定的支撑技术 ,但是它在蛋白质间相互作用分析及结构与功能的关系分析方面显得无能为力 ,而生物分子相互作用分析技术 (Ｂｉｏｍｏｌｅｃｕｌａｒｉｎｔｅｒａｃｔｉｏｎａｎａｌｙｓｉｓ ,ＢＩＡ)是进行蛋白质相互作用分析的较好技术 ,将两者结合起来在蛋白质组…     

常见生物传感芯片及其应用进展

生物传感芯片是一类综合了生物芯片和生物传感器的优点的新型生物芯片,在保持传统生物芯片的高通量、可寻址、并行处理等特点的基础上,与生物传感器技术相结合,进一步提高了芯片检测的灵敏度和特异性。常见的生物传感芯片主要有光纤传感芯片、表面等离子体共振传感芯片、热生物传感芯片、压电晶体传感芯片等,可用于各种生物大分子,如蛋白质、核酸等的检测,金属离子的测定,病原体的检测,药物筛选等。     

SPR生物传感器的特点及其在生物特异性相互作用分析中的应用

利用表面等离子模共振技术（SPR）进行生物特异性相互作用分析（BIA）已成为现代基因工程技术中的一种先进的手段。与传统的研究方法如酶联免疫吸附测定（ELISA）相比,它具有方便快捷、灵敏度高,应用范围广,实时监控等多项特点,利用这种新型研究手段对于生命科学的基础研究。医学诊断以及治疗等方面有着十分重的意义。本粗略概括了近几年来利用SPR生物传感器进行基础研究的基本情况以及对其的改进,并简要分析了此项技术的优点以及发展前景。     

表面等离子体共振技术在分子生物学中的应用

表面等离子体共振(SPR)技术可以实时、原位地测定生物分子间的相互作用而无需任何标记,可以连续监测吸附和解离过程,并可以进行多组分复合物的相互作用的研究。SPR技术在DNA的复制和转录、DNA的修复、核酸与药物的作用以及肽库和抗体库的筛选等分子生物学领域的应用研究取得了令人瞩目的进展,显示了常规技术无法比拟的优越性。     

基于SPR生物传感器的免疫学检测

利用一种全新的生物大分子相互作用检测仪表面等离子激元共振（SPR）生物传感器,对乙肝表面抗原,抗体,破伤风类毒素,抗体等生物制品进行生物特异性相互作用分析（BIA）,并对其在免疫学检测上的特征进行了探讨。     

偏振干涉角度调制SPR传感技术研究及应用进展

针对一种新兴生物检测方法——表面等离子体波共振(SPR)技术,文中SPR传感系统采用偏振干涉和角度调制方案,使SPR传感灵敏度与光复反射系数的模和相位都相关,从而实现较大线形范围内的高灵敏测量。同时开展了该SPR传感系统在环保领域的应用研究,SPR共振信号可实时随甲烷含量线性改变,气体检测灵敏度达到1 070ppm,实验结果验证了这种SPR传感技术的检测性能并显示了其在环保监测领域的应用潜力。     

SPR蛋白质芯片在输入性疟疾筛查中的应用

输入性疟疾已是我国疟疾防控的主要危险因素,如何对入境人员进行疟疾快速筛查是急需解决的难题。蛋白质芯片已被广泛应用于高通量筛选和诊断,本研究尝试构建了表面等离子共振技术 (Surface plasmon resonance,SPR) 蛋白芯片用于恶性疟疾的快速检测。采用聚乙二醇高分子处理的特异性吸附表面,以恶性疟疾特异性抗原富组氨酸蛋白Ⅱ (Histidine-rich protein Ⅱ,HRP2) 作为捕获探针,建立疟疾的微阵列芯片,并对芯片的最佳抗原固定浓度,检测的灵敏性和特异性,以及抗干扰能力进行了分析。该芯片可成功应用于恶性疟疾的筛查,具有无标记、即时快速的特点,与荧光定量PCR法相比,两种方法在敏感度和特异性方面无统计学差异。研究结果为一步研制疟疾分型鉴定蛋白质芯片奠定了基础,有利于对入境人员进行疟疾快速筛查。     

基于波长寻址的表面等离子体共振传感技术

本文提出了基于光谱扫描技术的非机械扫描的表面等离子体共振(SPR)传感技术,采用白光为SPR激发光源,通过单色仪控制入射光的波长实现光谱寻址,在保证灵敏度和动态范围的同时,使系统在整个动态范围内具有较好的线性,简化了传感器结构。理论分析了光谱扫描SPR传感技术的灵敏度和动态范围,搭建了实验系统,并测量了不同浓度的酒精水混合溶液的SPR信号变化。结果表明:系统折射率测量范围为1.30-1.38,灵敏度可达3.1×105RIU。     

SPR studies of carbohydrate-lectin interactions as useful tool for screening on lectin sources

Lectins are proteins or glycoproteins from plants, animals or microorganisms, which typically bind specifically to sugar residues, e.g., located in cell walls or membranes. This reaction might change the physiology of the cell wall and influences the metabolism inside the cell. Some lectins of plants stimulate the immune system by unspecific activation of T-cells or influence cell division; others cause agglutination of cells (e.g., erythrocytes) and are therefore from therapeutic interest.

In a new approach, biomolecular interaction analysis (BIA) was utilised for a screening program on lectins. The BIA has been done by surface plasmon resonance (SPR). The system can be used either for characterisation of lectin-binding domains or for a screening on lectins from natural sources. Several lectin-binding surfaces on the basis of SPR have been established.     

Real-Time, label-free monitoring of tumor antigen and serum antibody interactions

Conventional techniques for the detection of biomolecular interactions can be limited by the need for exogenous labels, time- and labor-intensive protocols, as well as by poor sensitivity levels. A refractometer instrument has been reconfigured to detect biomolecular interactions through changes in surface plasmon resonance (SPR). The binding kinetics and affinity values of anti-NY-ESO-1 monoclonal antibody, ES121, to the cancer-testis antigen NY-ESO-1 were determined according to the surface heterogeneity model and resulted in K(D) values of 1.3x10(-9) and 2.1x10(-10) M. The reconfigured instrument was then used to measure the interaction between tumor antigens and serum antibodies against these antigens in preselected cancer patient sera samples. The tumor antigens assayed included NY-ESO-1, SSX2 and p53, all used as recombinant proteins containing polyhistidine tags. These results demonstrated that the instrument is capable of detecting the binding of serum antibodies from cancer patient sera to immobilized tumor antigens, consistent with those observed previously in ELISA-based experiments. These results demonstrate the potential of SPR technology for the rapid diagnosis and monitoring immune responses.     

Biosensor detection of triplex formation by modified oligonucleotides

Due to the instability of DNA oligonucleotides in biological solutions, antisense or antigene therapies aimed at modulation of specific gene expression will most likely require the use of oligonucleotides with modified backbones. Here, we examine the use of a surface plasmon resonance biosensor (BIAcore) to compare triplex-directed binding of modified oligonucleotides targeted to a region of the murine c-myc promoter. We describe optimization of experimental conditions to minimize nonspecific interactions between the oligonucleotides and the sensor chip surface, and the limitations imposed by certain backbones and sequence types. The abilities of pyrimidine oligonucleotides with various modified backbones to form specific triple helices with an immobilized hairpin duplex were readily determined using the biosensor. Modification of the third-strand oligonucleotide with RNA or 2(')-O-methyl RNA was found to enhance triplex formation, whereas phosphorothioate or phosphotriester substitutions abrogated it. A comparison of these results to DNase I footprinting experiments using the same oligonucleotides showed complete agreement between the two sets of data.     

基于表面等离子共振的新型生物传感技术及其在生命科学中的应用

生物分子的活性功能是通过分子之间的相互作用来体现的,了解这种相互作用的过程对于生命科学领域的研究及揭示生命发生发展的基本机制具有重要的意义。基于表面等离子共振(surface plasmon resonance,SPR)的新型生物传感技术——BIAcore(biomolecular interaction analysis)是研究生物分子相互作用的理想工具。它可以实时跟踪检测生物分子间结合、解离的整个过程,已被广泛应用于蛋白质组学、信号转导、新药开发、遗传学分析和食品检测等领域,并且显示出广阔的应用前景。     

Biophysical characterization of glycosaminoglycan-IL-7 interactions using SPR

Glycosaminoglycans (GAGs) interact with a number of cytokines and growth factors thereby playing an essential role in the regulation of many physiological processes. These interactions are important for both normal signal transduction and the regulation of the tissue distribution of cytokines/growth factors. In the present study, we employed surface plasmon resonance (SPR) spectroscopy to dissect the binding interactions between GAGs and murine and human forms of interleukin-7 (IL-7). SPR results revealed that heparin binds with higher affinity to human IL-7 than murine IL-7 through a different kinetic mechanism. The optimal oligosaccharide length of heparin for the interactions to human and murine IL-7 involves a sequence larger than a tetrasaccharide. These results further demonstrate that while IL-7 is principally a heparin/heparan sulfate binding protein, it also interacts with dermatan sulfate, chondroitin sulfates C, D, and E, indicating that this cytokine preferentially interacts with GAGs having a higher degree of sulfation.     

Aptamer–Au NPs conjugates-enhanced SPR sensing for the ultrasensitive sandwich immunoassay

The goal of this work is to explore the amplification effect of aptamer–gold nanoparticles (Au NPs) conjugates for ultrasensitive detection of large biomolecules by surface plasmon resonance (SPR). A novel sandwich immunoassay is designed to demonstrate the amplification effect of aptamer–Au NPs conjugates by using human immunoglobulin E (IgE) as model analyte. Human IgE, captured by immobilized goat anti-human IgE on SPR gold film, is sensitively detected by SPR spectroscopy with a lowest detection limit of 1 ng/ml after anti-human IgE aptamer–Au NPs conjugates is used as amplification reagent. Meanwhile, the non-specific adsorption of aptamer–Au NPs conjugates on goat anti-human IgE is confirmed by SPR spectroscopy and then it is minimized by treating aptamer–Au NPs conjugates with 6-mercaptohexan-1-ol (MCH). These results confirm that aptamer–Au NPs conjugates is a powerful sandwich element and an excellent amplification reagent for SPR-based sandwich immunoassay.     

A novel aldehyde dextran sulfonate matrix for affinity biosensors

Aldehyde dextran sulfonate (ADS), a modified oligosaccharide polymer, was used to prepare a new matrix structure for affinity biosensors. The principal difference between the ADS matrix and similar structures developed previously results from presence of two active functional groups in the matrix, namely, aldehyde and sulfonate. These groups perform two different functions in the matrix. The aldehyde group is responsible for covalent bonding in the biomaterials, and the negatively charged sulfonate group provides electrostatic attraction of the positively charged biomolecules. By varying the ratio between the aldehyde and sulfonate groups in the matrix, one can control contributions from the two binding modes (covalent and electrostatic). A number of oligosaccharides, such as simple dextran, aldehyde dextran (AD), aldehyde dextran sulfonate (ADS) and aldehyde ethylcellulose (AEC), were used for preparation of matrix structures. The properties of the obtained matrices were analysed and compared. Surface plasmon resonance (SPR) was used as the main technique to characterize the matrix structures.