Hyphenated LC-MS Platform for Characterization of Low Abundance Proteins with Pos

描述（由申请人提供）：开发新的分析工具和方法以可靠地鉴定和定量蛋白质翻译后修饰（PTM）（如磷酸化）至关重要。现有的方法大多是基于质谱与液相色谱（LC-MS）的组合。LC起着分离平台的作用，尽管其分析能力可以增加有关蛋白质化学和物理性质的宝贵信息。本项目的目的是开发一个商业上可行的分析平台，将整合两个序列相关的数据流从LC和串联质谱组合。该平台的主要特点是将PTM表征LC-MS硬件和一套开发的软件工具相结合，利用新型生物大分子临界条件液相色谱（BioLCCC）技术预测PTM肽的序列依赖性保留时间。这种新方法有望提高修饰蛋白分析的通量、分析物种的动态范围，并允许快速、明确地鉴定沿着蛋白质氨基酸序列的修饰位点。 公共卫生相关性：蛋白质组学工具的应用在现代基础科学、药物发现和临床应用中发挥着重要作用。我们提出了一个新的平台，磷蛋白鉴定使用液体分离，精细富集技术，和多模式串联质谱，以提高蛋白质组学的可靠性和吞吐量。

DESCRIPTION (provided by applicant): It is vitally important to develop novel analytical tools and approaches for reliable identification and quantitation of protein posttranslational modifications (PTMs) such as phosphorylation. The existing approaches are mostly based on the combination of mass spectrometry with liquid chromatography (LC-MS). LC plays a role of a separation platform although its analytical capabilities can add an invaluable piece of information about chemical and physical properties of proteins. The purpose of this project is to develop a commercially viable analytical platform that will integrate two sequence related data streams from LC and tandem mass spectrometry combination. The main feature of the proposed platform, which will include a combination of PTM characterization LC-MS hardware and a set of developed software tools, is the utilization of the novel Liquid Chromatography of Biomacromolecules at Critical Conditions (BioLCCC) technology for sequence-dependent retention time prediction for peptides with PTMs. The new approach is expected to increase the throughput of modified protein analysis, the dynamic range of analyzed species, and to allow rapid, unambiguous identification of the sites of modifications along the protein amino acid sequences. PUBLIC HEALTH RELEVANCE: The application of proteomics tools plays an important role in modern basic science, drug discovery and clinical applications. We propose a new platform for phosphoprotein identification using liquid separation, refined enrichment technique, and multimode tandem mass spectrometry to increase reliability and throughput in proteomics.