Summary

The analysis of biological fluids to identify proteins that may indicate a disease setting, state and progression, is an increasingly explored field. Despite the expectatives created, there are several hurdles that must be solved to reach an extensive proteome coverage using mass spectrometry, mainly due to the complex composition of the matrices. In this regard, bile is specially challenging and yet, very attractive, as a proximal fluid that might provide valuable information for the management of liver and pancreas associated diseases. Proteins account for less than 5% of bile organic components and, although optimized protocols for protein extraction have been developed, only partial descriptions of bile proteome have been achieved. In this manuscript a new procedure is described that significantly improves protein recovery from rat bile, which reduces by a factor of six the sample amount required for a typical proteomics analysis. Moreover, the number of proteins reliably identified in a single nanoLC-MS/MS run from 1 μg protein was increased by three-fold. This procedure provides a valuable resource to dig deeper into the molecular composition of bile and open new avenues to identify new hallmarks of disease such as cholangiocarcinoma, hepatocellular carcinoma and pancreatic cancer for their better clinical management.

中文翻译：

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深入研究胆汁蛋白质组。

概要

对生物流体进行分析以鉴定可能指示疾病状况，状态和进展的蛋白质的研究越来越多。尽管产生了期望值，但由于基质的复杂组成，使用质谱仪达到广泛的蛋白质组覆盖率仍需要解决几个障碍。在这方面，胆汁作为一种近端液体特别具有挑战性，但是非常有吸引力，它可以为管理肝脏和胰腺相关疾病提供有价值的信息。蛋白质占胆汁有机成分的比例不到5％，尽管已开发出优化的蛋白质提取方案，但仅获得了部分胆汁蛋白质组的描述。在本手稿中，我们描述了一种新的程序，该程序可以显着提高大鼠胆汁中蛋白质的回收率，这将典型蛋白质组学分析所需的样品量减少了六倍。此外，在单个nanoLC-MS / MS中从1μg蛋白质中可靠鉴定出的蛋白质数量增加了三倍。该程序提供了宝贵的资源，可更深入地研究胆汁的分子组成并开辟新途径，以鉴定诸如胆管癌，肝细胞癌和胰腺癌等疾病的新特征，以更好地进行临床管理。