SBIR Phase I: Universal Proteome Sample Preparation Kit Development

这个小企业创新研究（SBIR）第一阶段项目的更广泛的影响/商业潜力是开发一种蛋白质样品制备试剂盒，消除变异性，并为研究人员提供他们需要的信息，以找到重要的蛋白质药物靶标和生物标志物。 蛋白质组学的高通量蛋白质分析方法正在对生物技术和制药行业产生重大影响，从基于蛋白质的新药开发到质量控制监测和生物标志物发现到临床诊断。所有蛋白质组学应用都需要稳健、可重复和可自动化的样品制备方法。然而，目前这些方法还没有标准化。缺乏标准化往往导致实验失败和不可靠的结果，这可能需要许多轮的实验重复，花费时间和金钱。标准化的蛋白质样品制备试剂盒将大大节省工作流程，缩短上市时间，并加快生物标志物的发现和药物开发。SBIR第一阶段项目的智力价值是开发一种通用的肽分离试剂盒，以满足蛋白质组学社区的不同需求。 目前，样品制备方法通常有十几个步骤，容易出现肽损失和偏倚，需要一天以上才能完成。 通过NSF I-Corps计划，发现学术界和工业界超过70%的研究人员对当前的蛋白质样品制备方法不满意。 为了满足这一需求，开发了一种提高肽样品制备速度和效率的样品制备试剂盒。这种方法与目前的方法在三个基本方面不同。 首先，所有操作都将在单个试管中的溶液中完成，与现有方案相比，步骤数量减少一半。第二，采用快速、通用和可逆的化学标签，而不是依赖于物理分离，防止样品损失或偏差。第三，稳定的胰蛋白酶衍生物可以完全消化蛋白质样品，然后有效去除完整和碎片的胰蛋白酶，防止污染。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值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 I project is to develop a protein sample preparation kit that eliminates variability and provides researchers with the information they need to find important protein drug targets and biomarkers. The high throughput protein analysis methods of proteomics are having a significant impact on the biotechnology and pharmaceutical industries, from the development of new protein-based drugs to quality control monitoring and biomarker discovery to clinical diagnostics. All proteomics applications require robust, repeatable and automatable sample preparation methods. Currently, however, these methods are not standardized. The lack of the standardization often leads to experimental failure and unreliable results, which may require many rounds of experimental repeats, costing time and money. Standardized protein sample preparation kits will provide significant workflow savings, improve time to market, and accelerate biomarker discovery and drug development.The intellectual merit of this SBIR Phase I project is to develop a universal peptide isolation kit to serve the diverse needs of the proteomics community. Currently, methods for sample preparation typically have more than a dozen steps, are prone to peptide loss and bias, and require more than a day to complete. Through the NSF I-Corps program, it was discovered that more than 70% of researchers in academia and industry are unhappy with current protein sample preparation methods. To address this need, a sample preparation kit that increases the speed and efficiency of peptide sample preparation was developed. The approach differs from current methods in three fundamental ways. First, all manipulations will be done in solution in a single tube with half the number of steps compared to existing protocols. Second, a fast, universal, and reversible chemical tag is employed rather than relying on physical separation, preventing sample loss or bias. Third, a stabilized derivative of trypsin will allow for complete digestion of protein samples followed by efficient removal of intact and fragmented trypsin, preventing contamination.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.