SBIR Phase I: Universal Proteome Sample Preparation Kit Development

这个小型企业创新研究(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，该试剂盒消除了变异性，并为研究人员提供他们寻找重要的蛋白质药物靶标和生物标志物所需的信息。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的开发到质量控制监测和生物标记物的发现，再到临床诊断。所有蛋白质组学应用都需要健壮、可重复和自动化的样品制备方法。然而，目前这些方法并不规范。缺乏标准化往往会导致实验失败和结果不可靠，这可能需要多次实验重复，耗费时间和金钱。标准化的蛋白质样品制备试剂盒将显著节省工作流程，缩短上市时间，并加快生物标记物的发现和药物开发。SBIR第一阶段项目的智力优势是开发一种通用的多肽分离试剂盒，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，容易出现多肽丢失和偏差，需要一天以上的时间才能完成。通过NSF I-Corps项目发现，超过70%的学术界和工业界研究人员对目前的蛋白质样品制备方法感到不满。为了满足这一需求，开发了一种提高多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有所不同。首先，与现有方案相比，所有操作都将在一个管子中完成，步骤数减少了一半。其次，使用快速、通用和可逆的化学标签，而不是依赖物理分离，防止样品丢失或偏差。第三，稳定的胰酶衍生物将允许完全消化蛋白质样品，然后有效地去除完整和破碎的胰酶，防止污染。这一奖项反映了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 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.