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

这个小型企业创新研究(SBIR)第一阶段项目将开发一种生物传感器来检测蛋白质或生物大分子(病毒、孢子等)。与当前最先进的技术相比，具有更高的特异性和更少的背景。目前，生物传感器依赖于固态、比色或荧光分析，这些分析需要昂贵的设备来进行结果评估。在这项提议中，这项技术是基于一种生物活性的、可光激发的纳米探针，该探针将在与感兴趣的目标结合时触发报告酶的激活。在这个第一阶段项目中，将基于相同的生物共轭纳米探针(BNP)平台开发两个生物传感器--一个表面传感器和一个溶液传感器。这些BNP传感器系统通过利用对感兴趣目标的多层检测特异性来改进夹心免疫分析的设计。只有当多个结合分子同时结合时，报告信号才会释放，但如果只有一个结合分子结合或没有结合分子结合，则阻止报告信号释放。这种设计允许目标分子的不同异构体或物种之间的区分，以及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先进水平。在这个第一阶段的项目中，活性生物共轭纳米探测器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，数字结果分别显示在LCD屏幕上或发送回监测点。由于可以用分光光度计询问结果，或者简单地以“是”或“否”的视觉结果来询问结果，因此可以使用溶液传感器来代替酶联免疫吸附测定，从而减少了对培养皿的需要。在商业上，这些生物传感器将代表着在检测化学和生物战以及简易爆炸装置毒剂方面的重大进步，因为它们的设计提高了严格程度，允许在发生绑定时进行实时检测，无需专门的检测设备即可报告结果，并允许远程审问。传感器的设计将区分代理的类型，并可靠地破译混合的代理，最终使用芯片上的细分平台来同时实现对多个目标的检测和区分。本提案中提出的生物传感器平台代表着检测技术的重大进步。这项研究应该会在国防部、国土安全部、生物技术和制药行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.