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LABEL-FREE DETECTION OF ANTIGENS USING IMPLANTABLE SERS NANOSENSORS

机译:使用可植入SERS纳米传感器的无标签抗原检测

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Monitoring the presence, production and transport of proteins inside individual living cells can provide vital information about cellular signaling pathways and the overall biological response of an organism. For example, cellular response to external stimuli, such as biological warfare (BW) agents, can be monitored by measuring interleukin-II (IL-2) expression inside T-cells as well as other chemical species associated with T-cell activation. By monitoring such species, pre-symptomatic detection of exposure to BW agents can be achieved, leading to significantly increased post-exposure survival rates. To accomplish such monitoring, we have developed and optimized implantable nanosphere-based nanosensors for the intracellular analysis of specific proteins in a label-free fashion. These sensors consist of 300-520 nm diameter silica spheres that have been coated with silver and antibodies to allow for trace protein detection via surface enhanced Raman spectroscopy (SERS). They have been optimized for SERS response by evaluating the size of the nanospheres best suited to 632.8 nm laser excitation, as well as the various nanosensor fabrication steps (i.e., silver deposition process, antibody binding, etc.). During usage, the presence of the specific protein of interest is monitored by either directly measuring SERS signals associated with the protein and/or changes in the SERS spectrum of the antibodies resulting from conformational changes after antigen binding. In this work, human insulin was used as a model compound for initial studies into the sensitivity of these optimized nanosensors.
机译:监测单个活细胞内部蛋白质的存在,产生和运输可以提供有关细胞信号通路和生物整体生物学反应的重要信息。例如,可以通过测量T细胞内的白介素II(IL-2)表达以及与T细胞活化相关的其他化学物质来监测细胞对外部刺激(例如生物战(BW)剂)的反应。通过监测此类物种,可以实现症状前检测到BW药剂的暴露,从而显着提高暴露后生存率。为了完成这种监视,我们已经开发和优化了可植入的基于纳米球的纳米传感器,用于以无标记的方式对特定蛋白质进行细胞内分析。这些传感器由直径300-520 nm的二氧化硅球组成,这些二氧化硅球已涂有银和抗体,可通过表面增强拉曼光谱(SERS)检测痕量蛋白质。通过评估最适合632.8 nm激光激发的纳米球的大小以及各种纳米传感器的制造步骤(即银沉积过程,抗体结合等),对SERS响应进行了优化。在使用过程中,通过直接测量与蛋白质相关的SERS信号和/或抗原结合后构象变化导致的抗体SERS光谱变化,来监控目标蛋白的存在。在这项工作中,人类胰岛素被用作模型化合物,以初步研究这些优化的纳米传感器的灵敏度。

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