SBIR Phase II: Universal Proteome Sample Preparation Kit Development

这个小型企业创新研究(SBIR)第二阶段项目的更广泛的影响/商业潜力是开发一种通用的蛋白质样品制备试剂盒，该试剂盒可以从任何生物样品中提取蛋白质，去除污染物，并为下游分析方法做准备，比现有技术具有更高的产量和更深的覆盖范围。蛋白质是细胞的分子机器，完成保持人类健康所需的所有工作，结合蛋白质分析对于发展对集成生物系统的清晰理解、实现精确医学的前景以及快速药物发现正变得越来越重要。样品制备是任何基于蛋白质的项目工作流程的第一步，如果没有一种高产、快速和可重复的方法来制备样品，下游分析就很难解释，而且变化无常，导致项目时间长和资源浪费。拟议的创新提高了样本产量、重复性和速度，加快了寻求发现新疗法和诊断方法的研究，并提高了成本效益。它还将从同一样本中提纯其他重要的生物分子，如DNA、RNA和代谢物，使研究人员能够从每个样本中获得更多信息，在准备一个样本所需的时间内准备多个样本，并通过使用相同的起始材料提高重复性。拟议的项目将利用可逆的蛋白质标记化学来创建一管工作流程，用于捕获、洗涤和洗脱用于分析的蛋白质样本。由于这种新型的可逆蛋白质标记化学不依赖于催化剂，不产生副产物，并且完全双正交，因此它具有与所有类型的缓冲系统通用的潜力，包括通常用于细胞裂解和蛋白质增溶的苛刻变性剂和洗涤剂。这项拟议的工作将优化这种样品制备技术在最常用的缓冲系统、模式生物中的利用，并将在血液、尿液和脑脊液等诊断样品类型的制备中显示出优异的产量。此外，由于我们的可逆蛋白质标签不会与其他生物分子发生交叉反应，我们将展示我们也可以使用这项技术来纯化其他重要的生物分子，如DNA、RNA和代谢物。该项目开发的技术将彻底改变研究人员准备分析样品的方式，为他们提供模块化工具，可以从相同的起始样品中提纯他们需要的任何生物分子，减少时间，提高产量，并通过在相同的起始样品中进行比较来减少批次之间的变异性。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is to develop a universal protein sample preparation kit that extracts proteins from any biological sample, removes contaminants, and prepares them for downstream analytical methods with higher yields and deeper coverage than currently available technology. Proteins are the molecular machines of the cell that do all the work required to keep humans healthy, and incorporation of protein analysis is becoming increasingly important for developing a clear understanding of integrated biological systems, realizing the promise of precision medicine, and rapid drug discovery. Sample preparation is the first step in any protein-based project workflow and without a high-yield, fast, and reproducible way to prepare samples, downstream analyses are difficult to interpret and highly variable, leading to long project times and wasted resources. The proposed innovation improves sample yields, reproducibility, and speed, accelerating research seeking to discover new therapies and diagnostics and improving cost-effectiveness. It will also purify other important biomolecules from the same sample, such as DNA, RNA, and metabolites, allowing researchers to obtain more information from each sample, prepare multiple samples in the time it takes to prepare one, and improve reproducibility by using the same starting material. The proposed project will utilize reversible protein tagging chemistry to create a one-tube workflow for the capture, wash, and elution of protein samples for analysis. Because this novel reversible protein tagging chemistry is not dependent on a catalyst, does not produce byproducts, and is completely biorthogonal, it has the potential to work universally with all types of buffer systems, including harsh denaturants and detergents commonly used for cell lysis and protein solubilization. This proposed work will optimize the utilization of this sample preparation technology in the most used buffer systems, model organisms, and will show superior yields in the preparation of diagnostic sample types such as blood, urine, and cerebrospinal fluid. Additionally, because our reversible protein tag does not cross react with other biomolecules, we will show that we can also use this technology to purify other important biomolecules such as DNA, RNA, and metabolites. The technology developed in this project will revolutionize how researchers prepare samples for analysis, providing them with modular tools that can purify any biomolecules they need from the same starting sample, reducing time, improving yields, and decreasing batch to batch variability by comparing within the same starting sample.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.