SBIR Phase I: Gas-Phase Two-Dimensional Electrophoresis

这个小型企业创新研究(SBIR)第一阶段项目将开发一种原型气相双向电泳仪(GP2DE)，与传统的二维凝胶电泳法(2DGE)相比，该仪器用于分析混合物中的蛋白质结构，样本量更少，吞吐量更高。该仪器利用之前开发的多维离子迁移率光谱(MDIMS)方法，通过高性能IMS在时间和空间上根据分子大小和形状分离气相蛋白质离子。了解蛋白质的功能对于实现个性化医学所需的进步至关重要，个性化医学将在科学和经济上有力地推动蛋白质组学领域的发展。因此，设计专门的疗法需要对完整的蛋白质结构有复杂的了解。GP2DE将提供一种快速工具，使用耦合离子迁移率光谱分析来检测蛋白质结构的差异，从而在几秒钟到几分钟内进行2D凝胶电泳型分离。提高蛋白质结构分析的吞吐量解决了在更快地建立结构知识库的劳动密集型2D GE过程中效率低下的问题。特别是，能够检测蛋白质翻译后修饰的工具，如糖基化或磷酸化，将对蛋白质组学有价值。GP2DE可以很容易地检测到这些结构变化，因为结构的不同会导致不同的离子迁移率。该项目的广泛影响/商业潜力如果成功，将为预算和/或设施空间有限的研究人员提供一种灵活的、通用的生物分子分析工具。这对于本科生实验室特别有用，因为缩短的学习曲线将允许在典型的时间和资源限制下进行更有成效的研究。所提出的气相双向电泳(G2DPE)是从先进的IMS技术发展而来的，用于结构尺寸和形状分析，以便于用户使用。未来五年，蛋白质组学研究产品的市场规模预计将达到数十亿美元。GP2DE减少了执行更耗时的2DGE分析的需要，这些分析具有更高的劳动力和供应运营成本，降低了研究成本，并提高了实验室生产率。因此，拥有另一种解释蛋白质结构信息的强大工具将推动更适合个人生理的下一代生物疗法的发展速度。开展的研究将在同行评议的科学期刊和科学会议上传播，以促进采用和推进蛋白质组学的多维IMS方法。

This Small Business Innovation Research (SBIR) Phase I project will develop a prototype gas-phase two-dimensional electrophoresis (GP2DE) instrument for analyzing protein structures in mixtures with less sample and higher throughput compared to traditional two-dimensional gel electrophoresis (2DGE). The instrument leverages the previously developed multi-dimensional ion mobility spectrometry (MDIMS) method for separating gas-phase protein ions based on molecular size and shape in both time and space with high-performance IMS. Understanding proteins' functions will be crucial to achieving the advances required for personalized medicine, which will be a strong driver of the proteomics field both scientifically and economically. Consequently, designing specialized therapies requires intricate knowledge of the complete protein structure. GP2DE will provide a rapid tool for detecting differences in protein structures using coupled ion mobility spectrometry analyses to give 2D gel electrophoresis-type separations in seconds to minutes. Increasing the throughput for analyses of protein structures addresses the inefficiencies in the labor intensive 2DGE process to build the structure knowledge base more rapidly. In particular, tools that can detect post-translational modifications of proteins, such as glycosylation or phosphorylation, will be valuable to proteomics. GP2DE can detect these structural changes easily because the structural differences will result in different ion mobilities. The broader impact/commercial potential of this project, if successful, will provide a flexible, general purpose biomolecule analysis tool for researchers with limited budgets and/or facility space. This would be particularly useful for undergraduate laboratories where the shortened learning curve would allow for more productive research under typical time and resource constraints. The proposed gas-phase two-dimensional electrophoresis (G2DPE) evolves from advanced IMS technologies for structural size and shape analyses that are geared to be user-friendly. The market for proteomics research products has been projected to be several billion dollars in the next five years. GP2DE reduces the need to perform many more time consuming 2DGE analyses that have higher operating costs from labor and supplies, decreasing research costs and adding to laboratory productivity. Therefore, having another powerful tool for elucidating protein structural information will propel the rate of development of the next-generation of biotherapies better tailored to individual physiologies. The research conducted will be disseminated in peer-reviewed scientific journals and at scientific conferences to foster the adoption and advancement of multi-dimensional IMS methods for proteomics.