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表面等离子体共振技术在分子生物学中的应用 总被引:6,自引:0,他引:6
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。 相似文献
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利用表面等离子体共振仪检测黄瓜花叶病毒 总被引:1,自引:0,他引:1
目的:研究一种便捷、高效地检测黄瓜花叶病毒(CMV)的方法。方法:利用表面等离子体共振(SPR)技术检测CMV。首先用11-MUA修饰SPR金片,再用EDC/NHS活化,之后通过NHS酯基与CMV抗体结合,用BSA封闭未结合的NHS酯基。将SPR金片装入SPR仪,通入待检样品,通过折射率变化实时监测实验过程。结果:该方法检测CMV的灵敏度能够达到10ng/mL,具有良好的特异性,与同属的花生矮化病毒、番茄不孕病毒无交叉反应。结论:建立的SPR方法操作简单、灵敏度高、特异性好,是一种新的高效检测CMV的方法。 相似文献
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表面等离子体激元共振与生物分子相互作用分析 总被引:6,自引:0,他引:6
对表面等离子体激元共振(surface plasmon resonance, SPR)的原理和在生物学研究方面的应用进行了综述.这种技术可以直接原位、实时地跟踪生物学实验研究系统,而不需要附加参数如进行标记等手段,具有高敏感性,也可以连续监测吸附或解吸附过程,目前有关的应用涉及到生物学结合分析、动力学及亲和力测定、免疫识别研究、结构与活性研究和核酸研究等多个领域. 相似文献
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表面等离子共振(surface plasmon resonance,SPR)技术旨在检测物体表面附近折射率的变化,其特点是无标记、实时、灵敏和快速,该技术多用于研究分子的相互作用,包括动力学、效率常数和大分子构象变化等。电化学(electrochemical,EC)技术是一项用于定性定量研究电子转移、物质氧化还原、界面吸附等过程的成熟技术,具有简单、低成本和设备小型化的优点。现有的DNA杂交技术,例如光学、电化学或压电转导技术,主要关注于提高DNA杂交检测系统的选择性和灵敏度。传统的SPR在DNA分析方面,由于无法测量折射率的极小变化而在超灵敏检测中的应用受到限制。因此,随着纳米材料的研发和联用技术的飞速发展,SPR与EC联用的生物传感器研究越来越成为人们关注的热点。近年来,关于SPR和EC联用在DNA检测方面的综述鲜有报道。对SPR和EC检测DNA的技术原理、联用方法、应用进展等方面作出了简要的介绍,以期为表面等离子共振和电化学联用的DNA传感器相关研究提供参考。 相似文献
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生物芯片技术在药物研究中的应用前景 总被引:3,自引:0,他引:3
1991年 ,美国Stephenfodor等提出了DNA芯片的概念[1] ,随着人类基因组计划 (HumanGenomeProject,HGP)的实施 ,生物芯片技术已成为基因组计划中的一种重要技术手段。生物芯片 (biologicalchip或biochip) ,把生化分析系统中的样品制备、生化反应和结果检测三个部分有机地结合起来连续完成。与传统的检测方法相比 ,具有高通量、高信息量、快速、微型化、自动化、成本低、污染少、用途广等特点[2 ] 。生物芯片包括基因芯片、蛋白质芯片或肽芯片、细胞芯片、组织芯片、元件型的微阵列… 相似文献
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基因药物是未来药物的发展方向,必须研究适宜的给药系统以促进基因药物的吸收和控制药效。基因药物的给药途径主要包括注射、口服、肺靶向、脑靶向和心血管基因转换等。所采用的剂型主要包括微球、脂质体、微乳等。本文就基因药物给药途径、所采用的主要剂型及各自所具备的优势作一综述。另外,简要介绍了基因药物给药系统的纳米技术。 相似文献
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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 总被引:5,自引:0,他引:5
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。 相似文献
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A. V. Kabashin V. E. Kochergin A. A. Beloglazov P. I. Nikitin 《Biosensors & bioelectronics》1998,13(12):1263-1269
A technique of phase-polarisation contrast (PPC) for the enhancement of the contrast of a surface plasmon resonance (SPR) intensity profile is proposed and experimentally realised. The technique exploits the peculiarities of light phase and polarisation behaviour under SPR. It applies to non-optimum SPR coupling conditions and enables one to lower the resonant minimum of reflected intensity nearly to zero, and hence to increase substantially the ratio of the intensity from the resonance to that at the minimum. We observed the contrast enhancement by more than one order of magnitude when we applied the PPC scheme. The PPC can be efficiently employed in commercial SPR sensors, as it significantly reduces restrictions on allowable parameters of SPR-supporting metal films and biomolecular layers immobilised on them, facilitates SPR observation, and increases the accuracy of SPR shift measurements. 相似文献
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Kibong Choi Heeju Youn Kwangjoong Kim Jungdo Choi 《Biotechnology and Bioprocess Engineering》1998,3(1):19-23
We enhanced the sensitivity of surface plasmon resonance biosensor by the conversion of the real-time direct binding immunoassay
into the sandwich immunoassay, in which colloidal gold particles coated with anti-mouse IgG was used. By the immobilization
of anti-mouse IgG onto the carboxymethyl dextran surface of thin gold film, the direct binding of analyte (mouse IgG) onto
the sensor chip, and the injection of colloidal gold particles coated with antimouse IgG, about 100 times of sensitivity enhancement
was obtained. This result suggests that nanoparticles, which has a high refractive index, homogeneous ultrafine structure
and capability of size control, would be applicable for the detection of very small quantity of biomaterial. 相似文献
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Surface plasmon resonance (SPR) technique was used to directly detect an intact form of insect pathogen: the baculovirus, Autographa californica multiple nuclear polyhedrosis virus (AcMNPV). An SPR sensor chip with three bio-functional layers was used to detect the intact AcMNPV: amine-reactive crosslinker with a disulfide bond that chemisorbs to gold film, Protein A, and a mouse IgG monoclonal antibody raised against a surface protein of the target viral pathogen. A two-channel (reference & test) micro-fluidic SPR system is used for reliable measurement. Bio-specific response to the AcMNPV is compared with the response for tobacco mosaic virus (TMV) as control. Successive exposure of the sensor chip to both viruses verifies a specific response to AcMNPV. This serves as a prerequisite to the development of a new type of viral pathogen detection sensors. 相似文献
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A surface plasmon resonance (SPR) sensor probe with integrated reference surface is described. In order to fabricate the integrated reference surface, two dielectric layers with different thickness were deposited on the single gold SPR sensor surface via plasma polymerization of hexamethyldisiloxane. The working sensor surface was a 34 nm dielectric layer with immobilized bovine serum albumin (BSA) antigen and an adjacent thin 1 nm dielectric layer without BSA provided reference surface. A specific immunoreaction of anti-BSA antibody was detected after immersion of the SPR probe into sample solution. Simultaneous observation of reference and working surface response enabled determination of the immunoreaction without the need for the baseline measurement. Moreover, compensation of nonspecific adsorption could be confirmed using anti-human serum albumin antibody. 相似文献
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The determination of equilibrium binding constants is an important aspect of the analysis of protein-protein interactions. In recent years surface plasmon resonance experiments (e.g., with a BIAcore instrument) have provided a valuable experimental approach to determining such constants. The standard method is based on measuring amounts of analyte bound at equilibrium for different analyte concentrations. During the course of a typical surface plasmon resonance experiment the measured equilibrium levels for a given analyte concentration often decrease. This appears to be due to a loss of activity of the protein coupled to the sensor chip or other phenomena. The loss in signal can lead to an erroneous determination of the equilibrium constant. A data analysis approach is introduced that aims to compensate for the loss of activity so that its influence on the results of the experiments is reduced. 相似文献
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It is established that achieving higher binding affinities in carbohydrate-protein interactions requires multivalent presentations of the sugar ligands at the receptor binding site. Several inhibition, calorimetric, mass balance, and other studies have reiterated the beneficial effects of molecular level clustering of the sugar ligands for tight binding to the receptors. We have undertaken an effort to study the multivalent effects involving larger assemblies, represented by micelles, and their lectin interactions. The micelles were constituted with monomer bearing one- or two-sugar moieties at the monomolecular level and with varying the distances between the sugar moieties. Micellar aggregation studies and dynamic light scattering (DLS) studies afforded details of the aggregation numbers and the hydrodynamic diameters of various glycolipid (GL) micelles. The GL micelles were used as analytes of surface plasmon resonance (SPR) experiments on a lectin concanavalin A (Con A)-immobilized surface. SPR studies of the micelle-lectin interactions demonstrate that the ligand-receptor binding can be fit into the bivalent analyte model of interaction. Furthermore, micelles formed from two-sugar containing GLs are able to elicit favorable kinetic association rate constants in comparison to the micelles constituted with one-sugar containing GLs. The kinetic rate constants across the micelles and the effect of the sugar valencies in the GLs are discussed. 相似文献
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《Expert review of proteomics》2013,10(3):425-433
Due to the enormous complexity of the proteome, focus in proteomics shifts more and more from the study of the complete proteome to the targeted analysis of part of the proteome. The isolation of this specific part of the proteome generally includes an affinity-based enrichment. Surface plasmon resonance (SPR), a label-free technique able to follow enrichment in real-time and in a semiquantitative manner, is an emerging tool for targeted affinity enrichment. Furthermore, in combination with mass spectrometry (MS), SPR can be used to both selectively enrich for and identify proteins from a complex sample. Here we illustrate the use of SPR-MS to solve proteomics-based research questions, describing applications that use very different types of immobilized components: such as small (drug or messenger) molecules, peptides, DNA and proteins. We evaluate the current possibilities and limitations and discuss the future developments of the SPR-MS technique. 相似文献
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Berit Johne 《Molecular biotechnology》1998,9(1):65-71
An epitope may be defined as a specific site on an antigen module characterized by the binding of one monoclonal antibody
(MAb). Epitope mapping by surface plasmon resonance in the BIAcore biosensor may be performed to characterize an antigen or
a group of specific MAbs or both. This article describes the BIAcore instrument and methods for such mapping. Examples include
molecular interaction studies with simple and complex proteins, such as myoglobin and calprotectin, respectively. 相似文献