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

这个小企业创新研究（SBIR）项目的更广泛的影响/商业潜力是开发技术，大大降低基于核糖核酸（RNA）的医学检测的成本和复杂性。 从生物样本中分离RNA是RNA诊断测试中的第一步，通常也是最困难的一步。从细菌到人类的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，有一个23亿美元的市场，用于清除RNA样品中的蛋白质和DNA污染。所提出的产品以近乎完美的选择性从样品中选择性地分离RNA，从而消除了对这些昂贵的RNA清除试剂的需求。RNA分离步骤的简化具有更广泛的影响，允许更小，更容易操作的设备，可以部署到当地诊所，允许更容易和更快的检测在床旁和农村设置。这个SBIR第一阶段项目提出开发一种新的碳基涂层的磁珠，选择性地分离RNA从样品中具有更好的产量比目前的产品，没有DNA或蛋白质污染。该公司已经开发出一种初步技术，在用于从培养细胞中分离RNA时，该技术的性能优于竞争产品。 在第一阶段项目中，材料特性和使用方案将得到优化，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。 将使用X射线光电子能谱（XPS）研究与本应用中的性能相关的材料特性，并进行优化，以最大限度地提高RNA对污染分子（如DNA和蛋白质）的选择性。RNA将从不同的生物物质（如软组织、血液和细胞培养物）中分离，并且该过程中的每个步骤都将进行优化，以最大限度地提高产量和RNA完整性（RIN），同时保持通过北方印迹、荧光光谱和qRT-PCR测定的完美纯度。该公司还将开始探索一种用于商业生产这种材料的制造系统。

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.