Mining Complex Human Fluids for New Potential Protein Biomarkers

 描述（由申请人提供）：在本提案中，我们试图证明一种新的实验方法，以开发一种新的消减方法，从复杂的人体液体样本中去除丰富的蛋白质。最终目标是开发一套方法或工具，以促进从头发现一套完整的低丰度潜在蛋白质生物标志物，从中可以进行更有针对性的研究。特别感兴趣的是发现癌症的潜在生物标志物，过去我们研究了肺癌样本和来自同一队列的参考文献。在本I期提案中，我们试图证明从人血浆中选择性和特异性清除人血清白蛋白亚型和二聚体。这些血清白蛋白包括人血清或血浆中蛋白质的一半，并且使用串联质谱法和差异凝胶电泳法两者引起非常真实的问题。第二阶段将扩展到去除三个数量级的丰富蛋白质，比现有的最先进的方法增加50倍。一个关键特征是选择条件以最小化蛋白质-蛋白质相互作用，这损害了现有方法的性能。所产生的简化的人体流体样品将与2D凝胶电泳和液相色谱-串联质谱法兼容。

 DESCRIPTION (provided by applicant): In this proposal we seek to demonstrate a new experimental approach to develop a new subtractive method to remove abundant proteins from complex human fluid samples. The ultimate goal is to develop a suite of methods or tools to facilitate the de novo discovery of a complete set of low-abundance potential protein biomarkers from which more targeted studies can be performed. A particular interest is in the discovery of potential biomarkers for cancers, and in the past we have studied lung cancer samples and references drawn from the same cohort. In this Phase I proposal, we seek to demonstrate the selective and specific removal of human serum albumin isoforms and dimers from human plasma. These serum albumins comprise half of the protein in human serum or plasma, and cause very real issues with both tandem mass spectrometry and difference gel electrophoresis approaches. The Phase II would extend to work to remove three orders of magnitude of abundant proteins, a 50 times increase from existing state-of-the-art methods. One key feature is that the conditions are chosen to minimize protein-protein interactions, which compromise the performance of existing methods. The simplified human fluid samples produced will be compatible with both 2D gel electrophoresis and liquid chromatography- tandem mass spectrometry approaches.