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用于监测神经炎症细胞因子血小板衍生生长因子的无试剂适体阻抗生物传感器。

Reagentless aptamer based impedance biosensor for monitoring a neuro-inflammatory cytokine PDGF.

作者信息

Liao Wei, Cui Xinyan Tracy

机构信息

Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA 15261, USA.

出版信息

Biosens Bioelectron. 2007 Sep 30;23(2):218-24. doi: 10.1016/j.bios.2007.04.004. Epub 2007 Apr 12.

Abstract

Neural prostheses often suffer from undesired chronic inflammatory tissue response. This can lead to neuronal loss and formation of glial scar tissue, which would serve as a barrier to neural signal transduction. In situ monitoring of neuro-inflammatory cytokines may improve our understanding of device induced inflammatory responses. Furthermore, early detection of the onset and degree of inflammation and releasing drugs accordingly may lead to improved long term performance of such implanted devices. For this reason, biosensor applying aptamer as probe and non-faradic electrochemical impedance spectroscopy (NIS) as the detection method has been developed. Aptamers, certain kinds of DNA or RNA molecules which can bind variety of molecules at high specificity, have the overwhelming advantages over antibodies of low cost and ease of use. Platelet-derived growth factor BB (PDGF-BB), one of the important cytokines involved in neural inflammation, has been selected as our detection target. Binding of PDGF to its aptamer immobilized on the silicon electrode surface lead to a decrease in capacitance measured by NIS. A good linear relationship between the decrease of capacitance and the logarithm of protein concentration was obtained, which proves the feasibility of quantitative measurements. By sweeping the applied electrode potential of potentiostatic EIS, -0.1 V to +0.1 V was determined to be the optimal range for achieving best discrimination between specific target binding and non-specific protein adsorption on aptamer-modified silicon surface. Under such conditions, the specificity of the detection measured by the ratio of the positive to negative control is around 10:1 and the detection limit is approximately 1 microg/ml (40 nM). The online measurement result exhibited negligible response for non-specific adsorption but significant signal changes for the specific target. Since the non-faradic strategy does not require any reagent to be loaded when performing the test, together with the ability of online measurements, this biosensor design is promising for in vivo monitoring.

摘要

神经假体常常遭受不良的慢性炎症组织反应。这会导致神经元丢失和胶质瘢痕组织形成,而胶质瘢痕组织会成为神经信号转导的障碍。对神经炎症细胞因子进行原位监测可能会增进我们对器件诱导的炎症反应的理解。此外,早期检测炎症的发生和程度并相应地释放药物可能会改善此类植入设备的长期性能。出于这个原因,已经开发出了以适体为探针、非法拉第电化学阻抗谱(NIS)为检测方法的生物传感器。适体是能够高特异性结合多种分子的某些种类的DNA或RNA分子,与抗体相比具有成本低和使用方便的压倒性优势。血小板衍生生长因子BB(PDGF - BB)是参与神经炎症的重要细胞因子之一,已被选作我们的检测靶点。PDGF与其固定在硅电极表面的适体结合会导致NIS测量的电容降低。电容降低与蛋白质浓度的对数之间获得了良好的线性关系,这证明了定量测量的可行性。通过扫描恒电位EIS的施加电极电位,确定-0.1 V至+0.1 V是在适体修饰的硅表面上实现特异性靶标结合与非特异性蛋白质吸附之间最佳区分的最佳范围。在这种条件下,通过阳性与阴性对照的比率测量的检测特异性约为10:1,检测限约为1微克/毫升(40 nM)。在线测量结果显示对非特异性吸附的响应可忽略不计,但对特异性靶标的信号变化显著。由于非法拉第策略在进行测试时不需要加载任何试剂,再加上在线测量的能力,这种生物传感器设计在体内监测方面很有前景。

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