SBIR Phase I: Active Titanium Dioxide Nanoprobes as Biosensors for Detection and Diagnostics

这项小企业创新研究（SBIR）第一阶段项目将开发一种生物传感器，用于实时检测蛋白质或生物大分子（病毒、孢子等），与目前最先进的技术相比，该传感器的特异性更高，背景值更低。目前，生物传感器依赖于固态、比色或荧光测定，需要昂贵的设备进行结果评估。在这项提议中，该技术是基于一种生物活性的、光激发的纳米探针，它将在与感兴趣的目标结合时触发报告酶的激活。在这个第一阶段的项目中，两个生物传感器——一个表面传感器和一个溶液传感器——将基于相同的生物共轭纳米探针（BNP）平台开发。这些BNP传感器系统通过利用对感兴趣的目标的多层检测特异性来改进三明治免疫分析的设计。报告信号仅在多个结合分子同时结合时被释放，但如果只有一个或没有结合分子被释放则被阻止释放。这种设计允许在目标分子的不同同种异构体或物种之间进行区分，并减少传感器中的背景噪声。具有这些属性的传感器将重新定义传感器和检测技术的最新技术。在这个第一阶段的项目中，活性生物共轭纳米探针将被开发成表面和溶液传感器。表面传感器可以用作本地或远程检测设备，其数值结果分别显示在LCD屏幕上或发送回监测站点。溶液传感器可用于代替ELISA测定，从而减轻了对平板仪的需求，因为结果可以用分光光度计查询或简单地作为“是”或“否”的视觉结果。在商业上，这些生物传感器将代表着探测化学和生物战以及简易爆炸装置药剂方面的重大进步，因为它们的设计提高了严格程度，可以在绑定发生时进行实时探测，无需专门探测设备就可以报告结果，并允许远程审讯。该传感器设计能够区分不同类型的智能体，并可靠地破译混合智能体，最终利用芯片上的细分平台实现对多个目标的同时检测和区分。本提案中提出的生物传感器平台代表了检测技术的重大进步。这项研究将在国防部、国土安全部、生物技术和制药工业以及学术界等不同领域产生理想的商业产品。

This Small Business Innovation Research (SBIR) Phase I project will develop a biosensor to detect proteins or biomacromolecules (viruses, spores, etc.) in real time with increased specificity and decreased background compared with the current state-of-the-art. Currently, biosensors rely on solid-state, colorimetric or fluorescence assays that require expensive equipment for result evaluation. In this proposal, the technology is based on a biologically-active, photoexcitable nanoprobe that will trigger the activation of a reporter enzyme upon binding to the target of interest. In this Phase I project, two biosensors -- a surface sensor and a solution sensor -- will be developed based on the same bioconjugated nanoprobe (BNP) platform. These BNP sensor systems improve on the design of the sandwich immunoassay by utilizing multi-layered detection specificity towards the target of interest. The reporter signal is released only when multiple binding molecules are engaged concurrently, but is prevented from release if only one or none of the binding molecules are engaged. This design allows for differentiation between different isoforms or species of the target molecule as well as decreases background noise in the sensor. A sensor with these attributes would re-define the state-of-the-art for sensor and detection technology. In this Phase I project active bioconjugated nanoprobes will be developed into both surface and solution sensors. The surface sensor could be used as a local or remote detection device, with numeric results appearing on an LCD screen or sent back to a monitoring site, respectively. The solution sensor could be used instead of an ELISA assay thereby alleviating the need for a platereader, as the results could be interrogated with a spectrophotometer or simply as a "yes" or "no" visual result. Commercially, these biosensors would represent significant advances in detection of chemical and biological warfare and improvised explosive device agents, as their design increases the level of stringency, allows detection in real time as binding occurs, reports results without specialized detection equipment, and allows remote interrogation. The sensor design would differentiate between type of agents and decipher a mixture of agents reliably, eventually using a subdivided platform on a chip to achieve detection and differentiation of multiple targets simultaneously. The biosensor platform presented in this proposal represents a major advance in detection technology. This research should result in desirable commercial products in areas as diverse as the Department of Defense, Homeland Security, the biotechnology and pharmaceutical industries, and academia.