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1.
在生命科学领域中,生物分子间的相互作用具有非常重要的作用。通过分子间相互作用分析不仅可阐明细胞生物学事件,而且为疾病发生机制和药物发现提供基础。MST技术是一种基于检测在温度梯度中的生物分子电泳迁移率的变化而检测生物分子间结合、解离过程,获取分子间相互作用的模式和动力学常数等方面信息的新技术,是近年来发展的研究生物分子相互作用的强有力工具,已广泛应用于生命科学领域研究。本文综述了MST的技术原理、分析方法及其在生命科学领域的应用进展。  相似文献   

2.
简要概述了生物电化学的研究领域,包括生物体系和生物界面模拟、生物分子的电化学、生物电催化、光合作用模拟和活组织电化学;总结了生物电化学传感器、生物芯片和生物电化学反应器在环境监测中的应用现状,并提出了其发展趋势,即不断向商品化方向发展,实现环境污染物的在线检测;利用基因技术,创造出检测能力更强的生物传感器和生物芯片;与其他精密分析仪器相结合,向多功能、集成化、智能化、微型化方向发展。  相似文献   

3.
生物分子的活性功能是通过分子之间的相互作用来实现的,了解这种相互作用的关系时生命科学的研究及揭示生命发生发展的基本机制具有着重要的意义.基于表面等离子共振(SPR)的分析分子相互作用(BIA)的技术是新型的生物传感技术,其无需标、能实时跟踪检测生物分子间结合、解离的整个过程,通过分析传感图谱获取分子相互作用的模式和动力学常数等方面的信息.SPR是研究生物分子相互作用的强有力工具,SPR技术已被广泛应用于生命科学领域的研究,并且显示出广阔的应用前景.概述了SPR技术原理、分析方法及其评述了其存在的问题.  相似文献   

4.
最近几年,生物传感器和流动注射、纳米技术等全新的技术进行结合,获得了前所未有的关注度,在未来也将会有无限的发展空间和可能性。基于生物分子之间的亲和性,亲和型生物传感器能够让生物活性之间创建全新的传感器装置,能够有很高的性能,例如特异性好,灵敏度高、成本花费少等优点,在生物医药学领域有很高的发展可能性。可以用于生物医学的标记物,对核酸、蛋自质等物质进行检测,研究药物作用机理,促进临床用药的筛选等相关工作。  相似文献   

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   

6.
生物正交化学反应是一类可以在生理条件下发生的化学反应,具有简单、高效、高特异性的特点,在生物医学的研究中被广泛应用.基于生物体天然生命过程的代谢工程,可对生物分子进行无损、高效的生物代谢修饰,是一种理想的生物修饰技术.通过生物代谢途径可有效地将各种化学报告基团引入靶标物的生物分子中,有利于携带配对基团的标记物与其发生生物正交反应,从而在活体系统中实现生物分子的标记示踪和药物递送.这种基于代谢工程与生物正交化学的标记策略因为具有两者之间的优势,在生物医学工程中的标记、成像示踪、诊断等领域展现出巨大的研究价值与应用潜力.本文介绍了生物正交和代谢工程的原理与生物医学研究进展,阐述了生物正交化学在分子成像和药物传递等方面的研究与应用.  相似文献   

7.
基于抗原-抗体识别的免疫分析技术在小分子化学性污染物监测领域占有重要地位,已成功应用于农药、兽药、生物毒素等的快速检测,为保障食品安全发挥了重要作用.但是,如何提高小分子半抗原的免疫原性及抗体的亲和力仍然是制约该领域发展的关键技术瓶颈.树状分子作为一类新型的高分子化合物,具有分子组成明确、结构规整、高度支化、纳米尺寸、单分散性以及表面呈现高密度功能团等众多优良的结构特性和良好的组织相容性,在小分子免疫分析领域具有潜在的应用优势.本文主要综述了树状分子作为载体在抗体制备及免疫分析方面的应用,重点介绍了树状分子作为载体在免疫原及包被原制备、作为免疫佐剂提高抗原免疫原性以及作为信号放大载体在提高免疫分析灵敏度等方面的研究现状,最后对其在小分子化学性污染物免疫检测领域的应用前景进行了评述,期望能为本领域研究人员提供借鉴.  相似文献   

8.
SPR技术在免疫学研究中的应用   总被引:1,自引:0,他引:1  
表面等离子共振(Surface Plasmon Resonance,SPR)技术是研究生物分子相互作用的强有力工具之一,该技术使生物分子之间相互作用的实时检测成为可能,并且灵敏度高、无需标记.通过分析传感图谱及分子相互作用的响应值获取分子相互作用的模式和动力学常数等方面的信息,并且获得的信息是能够定性和定量.SPR技术现在已广泛应用于生物、化学、免疫学研究及新药开发等领域.本文主要就SPR技术在免疫学研究中抗体活性检测、抗原表位预测等方面的应用进行了综述.  相似文献   

9.
弹性是生物分子网络重要且基础的属性之一,一方面弹性赋予生物分子网络抵抗内部噪声与环境干扰并维持其自身基本功能的能力,另一方面,弹性为网络状态的恢复制造了阻力。生物分子网络弹性研究试图回答如下3个问题:a. 生物分子网络弹性的产生机理是什么?b. 弹性影响下生物分子网络的状态如何发生转移?c. 如何预测生物网络状态转换临界点,以防止系统向不理想的状态演化?因此,研究生物分子网络弹性有助于理解生物系统内部运作机理,同时对诸如疾病发生临界点预测、生物系统状态逆转等临床应用具有重要的指导意义。鉴于此,本文主要针对以上生物分子网络弹性领域的3个热点研究问题,在研究方法和生物学应用上进行了系统地综述,并对未来生物分子网络弹性的研究方向进行了展望。  相似文献   

10.
蛋白质相互作用是生命活动中一种极其重要的生物分子关系, 对此领域的研究不仅具有理论意义, 还具有较强的应用价值. 近年来, 随着研究的深入, 各种蛋白质相互作用的生物医学文献激增, 挖掘其中的蛋白质相互作用关系成为人们面临的一大挑战. 当前, 已提出了多种文本挖掘方法, 对分散于生物医学文献中的蛋白质相互作用信息进行结构化或半结构化处理. 对这些工作进行分析, 总结出基于生物文本挖掘蛋白质相互作用信息的一般流程, 从蛋白质命名实体的识别、蛋白质相互作用关系的提取和蛋白质相互作用注释信息的提取3个子任务进行阐述, 同时介绍了生物文本挖掘领域的评测会议和一些挖掘蛋白质相互作用相关信息的工具. 最后, 对该领域存在的一些重要问题进行分析, 并预测了未来可能的发展方向, 以期对该领域相关研究提供一定的参考.  相似文献   

11.
The electron–electron double resonance (DEER) method, which provides distance distributions between two spin labels, attached site specifically to biomolecules (proteins and nucleic acids), is currently a well-recognized biophysical tool in structural biology. The most commonly used spin labels are based on nitroxide stable radicals, conjugated to the proteins primarily via native or engineered cysteine residues. However, in recent years, new spin labels, along with different labeling chemistries, have been introduced, driven in part by the desire to study structural and dynamical properties of biomolecules in their native environment, the cell. This mini-review focuses on these new spin labels, which allow for DEER on orthogonal spin labels, and on the state of the art methods for in-cell DEER distance measurements.  相似文献   

12.
Colloidal gold nanoparticles (AuNPs), with unique properties such as highly resonant particle plasmons, direct visualization of single nanoclusters by scattering of light, catalytic size enhancement by silver deposition, conductivity, and electrochemical properties, are very attractive materials for several applications in biotechnology. Furthermore, as excellent biological tags, AuNPs can be easily conjugated with biomolecules and retain the biochemical activity of the tagged biomolecules, making AuNPs ideal transducers for several biorecognition applications. The goal of this article is to review recent advances of using AuNPs as labels for signal amplification in biosensing applications. We focus on the signal amplification strategies of AuNPs in biosensing/biorecognition, more specifically, on the main optical and electrochemical detection methods that involve AuNP-based biosensing. Particular attention is given to recent advances and trends in sensing applications.  相似文献   

13.
Nanoparticle labels conjugated with biomolecules are used in a variety of different assay applications. In this paper, a sensitive fluoroimmunoassay for recombinant human interleukin-6 (IL-6) with the functionalized Rubpy-encapsulated fluorescent core-shell silica nanoparticles labeling technique has been proposed. IL-6 was measured based on the specific interaction between captured IL-6 antigen and functionalized fluorescent core-shell nanoparticles-labeled anti-IL-6 monoclonal antibody. The calibration graph for IL-6 was linear over the range 20-1250 pg ml(-1) with a detection limit of 7 pg ml(-1) (3 sigma). The regression equation of the working curve is I(F)=7.665+32.499[IL-6] (ng ml(-1)) (r=0.9980). The relative standard deviation (R.S.D.) for 11 parallel measurements of 78 pg ml(-1) IL-6 was 3.2%. Furthermore, the application of fluorescence microscopy imaging in the study of the antibody labeling and sandwich fluoroimmunoassay with the functionalized fluorescent core-shell silica nanoparticles was also explored. This proposed method has the advantage of showing the specificity of immunoassay and sensitivity of fluorescent nanoparticle labels technology. The results demonstrate that the method offers potential advantages of sensitivity, simplicity and reproducibility for the determination of IL-6, and is applicable to the determination of IL-6 in serum samples and enabling fluorescence microscopy imaging for the determination of IL-6.  相似文献   

14.
We report the development of phosphorylcholine (PC) group-covered nanoparticles for multiple immobilization reactions; the surface of these nanoparticles facilitates bioreactions such as enzymatic reactions and molecular diagnoses. The nanoparticles were covered with a bioconjugate PC group containing a polymer backbone, and their surface properties were as follows: (1) suppression of nonspecific protein adsorption and (2) stabilization of immobilized biomolecules. In this study, biomolecules were immobilized on PC-covered nanoparticles by using different spacer lengths between the polymer backbone and biomolecules. The stability of the immobilized biomolecules was evaluated using horseradish peroxidase-labeled IgG, and the bioconjugate nanoparticles were stored at 4, 25, and 40 °C. The residual enzymatic activity of the peroxidase was monitored at a particular time. On the other hand, to test the role of these nanoparticles in molecular diagnosis, we used IgG-conjugated nanoparticles and the fluorescence resonance energy transfer (FRET) phenomenon. The IgG molecules were labeled with either donor or acceptor molecules, and each labeled IgG was simultaneously immobilized on the PC-covered nanoparticles. These labeled IgG molecules induce the FRET phenomenon upon capture of the target antigen provided they are in close proximity. The resulting fluorescence was readable via the FRET phenomenon. In the present study, C-reactive protein (CRP) was used as the target antigen, and the effect of the spacer length is discussed. The bioconjugated nanoparticles covered with PC groups are promising tools for tuning bioreactions.  相似文献   

15.
A method using confocal Raman microspectroscopy for the detection of cellular proteins in single intact cells was developed. Two approaches were used to improve the detection of these cellular components. First, compounds with high Raman scattering were investigated for potential use as Raman labels. Raman labels were conjugated to either biomolecules or biotin and used as markers in the detection of cellular enzymes and receptors. Second, silver colloids were used to increase the surface-enhanced Raman scatter (SERS) of these Raman labels. Cresyl violet and dimethylaminoazobenzene are Raman labels that provide very sensitive SERS detection by a confocal Raman microscope with a HeNe laser at wavelength of 632.8 nm. The detection of 12-lipoxygenase and cyclooxygenase-1 in single bovine coronary artery endothelial cells and the binding of angiotensin II to its receptors in zona glomerulosa cells was demonstrated.  相似文献   

16.
Silver nanoparticles have been modified with self-assembled monolayers of hydroxyl-terminated long chain thiols and encapsulated with a silica shell. The resulting core–shell nanoparticles were used as optical labels for cell analysis using flow cytometry and microscopy. The excitation of plasmon resonances in nanoparticles results in strong depolarized scattering of visible light, permitting detection at the single nanoparticle level. The nanoparticles were modified with neutravidin via epoxide–azide coupling chemistry, to which biotinylated antibodies targeting cell surface receptors were bound. The nanoparticle labels exhibited long-term stability in solutions with high salt concentrations without aggregation or silver etching. Labeled cells exhibited two orders of magnitude enhancement of the scattering intensity compared with unlabeled cells.  相似文献   

17.
This paper presents the microstructured deposition of nanoparticles, which are chemically modified to be conjugated to biomolecules or dyes. In our approach, we separated the technical steps “surface chemistry” and “microstructuring” into two fully independent processes. This was realized by firstly synthesizing specific nanoparticles with tailor‐made surfaces. Thus, the surface chemistry was customized to the demands of most different kinds of specific proteins. Then, microstructured monolayer arrays of these nanoparticles were generated by photolithography, microcontact printing or spotting with a microarrayer. The overall system resulted in a nanoparticle‐based microarray for the selective binding of protein ligands providing both wide flexibility and high specificity.  相似文献   

18.
人体内各种复杂的生命活动离不开蛋白质之间的相互作用。这种相互作用具有瞬时性和结合力弱等特点,并受到多种动态调节,特别是蛋白质翻译后修饰(post-translation modifications, PTM)。传统的亲和质谱检测方法存在蛋白纯化的局限性,在高效检测到动态变化方面存在不足。邻近标记是一种能够给与靶蛋白质瞬时靠近,或者互作(邻近)的蛋白质加上生物素的技术,它与质谱检测技术的联合使用能检测细胞过程中弱的、瞬时的蛋白质相互作用,有效解决上述问题。本文综述了基于生物素的邻近标记方法的发展现状,从依赖于融合序列的生物素标记开始,依次介绍有关生物素连接酶、过氧化物酶及其进化后的2代标记方法等经典生物素标记的方法和原理,比较各个方法间的差异和优缺点;也列举了一些近年来新出现的标记方法,如将生物素连接酶进行拆分、鉴定蛋白质在不同复合物中功能的方法、抗体靶向的标记方法,以及其他来源的生物素连接酶突变体,例如枯草芽孢杆菌(Bacillus subtilis)的C端氨基酸突变的生物素连接酶,能够应用在苍蝇和蠕虫中的生物素连接酶突变体。本文对这些方法进行归纳总结,旨在为初步接触该领域的科研工作者提供参考,同时也希望能够提供一些新的思路,推动蛋白质相互作用组学的发展。  相似文献   

19.
p-Isothiocyanatophenyl derivatives of Pt(II)- and Pd(II)-coproporphyrin I are described as stable monofunctional reagents which enable simple covalent labeling of proteins and other biomolecules under mild conditions in aqueous solutions. Labeling procedure was optimized for antibodies, avidin, and neutravidin. Photophysical properties of resulting conjugates important for their use in binding assays based on time-resolved phosphorescence detection were studied. The functional activity and long-term storage stability of antibody conjugates were assessed in comparison with unmodified proteins. The new labels and their conjugates were evaluated in the solid-phase immunoassays using commercial time-resolved phosphorescence readers Victor(2) and Arcus-1230 (Wallac). Potential applications of these reagents in in vitro diagnostics are discussed.  相似文献   

20.
This paper describes the combination of electrochemical immunosensor using gold nanoparticles (GNPs)/carbon nanotubes (CNTs) hybrids platform with horseradish peroxidase (HRP)-functionalized gold nanoparticle label for the sensitive detection of human IgG (HIgG) as a model protein. The GNPs/CNTs nanohybrids covered on the glass carbon electrode (GCE) constructed an effective antibody immobilization matrix and made the immobilized biomolecules hold high stability and bioactivity. Enhanced sensitivity was obtained by using bioconjugates featuring HRP labels and secondary antibodies (Ab2) linked to GNPs at high HRP/Ab2 molar ratio. The approach provided a linear response range between 0.125 and 80 ng/mL with a detection limit of 40 pg/mL. The immunosensor showed good precision, acceptable stability and reproducibility and could be used for the detection of HIgG in real samples, which provided a potential alternative tool for the detection of protein in clinical laboratory.  相似文献   

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