Proteomic technologies for complex samples

细胞中的生物过程本质上是动态的，这种动态可能受到疾病和其他因素的影响。在蛋白质水平上，这种动态表现为特定蛋白质数量的变化以及这些蛋白质所携带的修饰。因此，识别和监测细胞中蛋白质水平及其修饰变化的能力可以极大地帮助我们了解生物过程，并可能为药物开发提供新的方法。不幸的是，目前还没有能够跟踪细胞中存在的蛋白质集合（蛋白质组）的技术。近年来，我们和其他研究小组致力于将蛋白质组分解成亚蛋白质组。我们已经证明，通过研究亚蛋白质组，可以分析以前从未观察到的蛋白质，并且可以使用简单的基于质谱的技术。我们建议开发一套基于亲和纯化和小型化的独立技术，这将允许亚蛋白质组的纯化和加工。特别是，我们将开发简单的二维和三维小型化系统，将微尺度技术与微尺度技术结合起来，针对特定的亚蛋白质组，然后处理该蛋白质组中的蛋白质。这些独立的系统将与许多实验室中现成的简单质谱技术兼容。我们建议建立定量亚蛋白质组蛋白水平的系统，并观察这些亚蛋白质组蛋白磷酸化的变化。我们设想，随着时间的推移，我们将拥有一系列这些亚蛋白质组分析系统，这些系统可以商业化，并用于提供更好的模型，增加对生物过程的理解，从而为药物开发提供新的方法。这将是蛋白质组学领域的一个重要发展，因为它将允许更多的蛋白质被识别和量化，并且它将很容易适应其他研究实验室。新技术的发展对加拿大非常重要，特别是那些可以商业化和可以直接应用于更好地了解人类疾病的技术。

Biological processes in cells are intrinsically dynamic and this dynamicity can be affected by diseases and other factors. At the protein level, the dynamicity is manifested by changes in the amount of specific proteins and in the modifications that these proteins carry. Therefore, the ability to identify and monitor the level of proteins in the cell and the changes in their modification can greatly help our understanding of biological process and potentially provide new approaches for drug development. Unfortunately, techniques that are capable of following the ensemble of proteins present in cells (the proteome) are currently not available. In recent years, we and other groups have focused on breaking the proteome into parts called sub-proteomes. We have demonstrated that by studying sub-proteomes, proteins never observed before can be analyzed and that simple mass spectrometry based techniques can be used. We propose to develop a suite of stand-alone technologies, based on affinity purification and miniaturization that will allow the purification and processing of sub-proteomes. In particular, we will develop simple two and three dimensional miniaturized systems that combine microscale techniques to target specific sub-proteomes with microscale techniques and then process the proteins in that proteome. These stand-alone systems will be compatible with simple mass spectrometry techniques readily available in many laboratories. We propose to build systems for quantitating protein levels in sub-proteomes and to look at the changes in protein phosphorylations in these sub-proteomes. We envision that over time we will have an array of these sub-proteome analytical systems that can be commercialized and used to provide a better model and increased understanding of biological processes resulting in new approaches for drug development. This will be an important development for the field of proteomics because it will allow more proteins to be identified and quantified, and it will be readily adaptable in other research laboratories. The development of novel technologies is of great importance to Canada, especially those that can be commercialized and can have direct application for better understanding human diseases.