SBIR Phase I: Highly Selective and Rapid Separation of RNA to Reduce Medical Diagnostic Costs

这一小型企业创新研究项目的更广泛影响/商业潜力是开发能够大幅降低基于核糖核酸（RNA）的医学检测成本和复杂性的技术。从生物样本中分离RNA是RNA诊断测试的第一步，也是最困难的一步。从细菌到人类，所有生物都受到病毒感染的威胁，并具有强大的天然防御能力；这些蛋白质防御对人类有益，但会干扰RNA检测。DNA污染也会干扰RNA检测。因此，用于清除RNA样本中蛋白质和DNA污染的试剂有23亿美元的市场。提出的产品有选择性地从样品中分离RNA，具有近乎完美的选择性，消除了这些昂贵的RNA清理试剂的需要。RNA分离步骤的简化具有更广泛的影响，允许在当地诊所部署更小、更容易操作的设备，允许在医疗点和农村环境中更容易、更快速地进行测试。这个SBIR一期项目建议开发一种新的碳基磁珠涂层，选择性地从样品中分离RNA，比现有产品的产量更高，而且没有DNA或蛋白质污染。该公司已经开发出一种初步技术，用于从培养细胞中分离RNA时，其性能优于竞争对手的产品。在这个第一阶段的项目中，材料特性和使用协议将被优化，以最大限度地提高实际应用中的性能，例如从血液中分离RNA。材料的特性，因为它们在此应用中的性能将使用x射线光电子能谱（XPS）进行研究，并优化，以最大限度地选择性RNA超过污染分子，如DNA和蛋白质。RNA将从软组织、血液和细胞培养物等多种生物物质中分离出来，过程中的每一步都将进行优化，以最大限度地提高产量和RNA完整性（RIN），同时通过northern blot、荧光光谱和qRT-PCR保持完美的纯度。该公司还将开始探索用于该材料商业化生产的制造系统。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project is to develop technology that will substantially reduce the cost and complexity of ribonucleic acid (RNA)-based medical testing. Isolation of RNA from biological samples is the first and often the most difficult step in RNA diagnostic testing. All living things from bacteria to humans are threatened by viral infection and have strong natural defenses; these protein defenses are beneficial to humans but interfere with RNA testing. DNA contamination also interferes with RNA testing. As a result, there is a $2.3B market for reagents to clean-up protein and DNA contamination in RNA samples. The proposed product selectively isolates RNA from samples with near perfect selectivity, eliminating the need for these expensive RNA clean-up reagents. Simplification of the RNA isolation step has the broader impact of allowing for smaller and easier to operate devices that can be deployed to local clinics, allowing for easier and faster testing in point-of-care and rural settings.This SBIR Phase I project proposes to develop a new carbon-based coating for magnetic beads that selectively isolates RNA from samples with better yield than current products and without DNA or protein contamination. The company has developed a preliminary technology that performs better than competing products when used to isolate RNA from cultured cells. During this Phase I project, the material properties and usage protocols will be optimized to maximize performance in real-world applications, such as isolation of RNA from blood. The material properties as they relate to performance in this application will be studied using x-ray photoelectron spectroscopy (XPS) and optimized to maximize selectivity for RNA over contaminating molecules such as DNA and proteins. RNA will be isolated from diverse biological substances such as soft tissue, blood, and cell cultures, and each step in the process will be optimized to maximize yield and RNA integrity (RIN) while maintaining perfect purity as determined by northern blot, fluorescence spectroscopy, and qRT-PCR. The company also will begin to explore a manufacturing system for commercial production of the material.