Burk Kyle, Legg Kevin, Danielson Phillip, Parker Glendon
Department of Environmental Toxicology, The University of California, Davis, Davis, CA, USA.
Protein Metrics, Cupertino, CA, USA.
Methods Mol Biol. 2025;2884:143-155. doi: 10.1007/978-1-0716-4298-6_10.
Biological fluids are proteinaceous liquids or suspensions released through different body orifices or through penetration of the skin. These fluids are the result of multiple tissues and cell types and contain extensive, highly complex, and dynamic protein populations that reflect both the transcriptional program of the originating cells and a record of the individual's health status. Body fluids are readily accessible to clinicians and researchers, and as such proteomic analyses are an important component of clinical studies, fertility studies, oral health studies, and forensic investigations. Current mass spectrometry (MS) datasets have a dynamic range of up to six orders of magnitude and are as diverse as the originating tissue types. Mass spectrometry has the potential to provide information across a wide range of applications, including basic research into human biology and pathology, biochemical analysis of protein function, biomarker discovery and detection, as well as forensic investigations wherein investigators interpret a protein profile to identify the body site origin of a biological fluid. The method below describes a specimen processing workflow that is flexible in terms of biological fluid type, sample state (e.g., a dried sample extracted from evidence or neat fluid), and level of degradation. The method described here is compatible with both high sensitivity shotgun liquid chromatography-mass spectrometry LC/MS analysis and targeted (qualitative or quantitative) MS-based analysis of biomarker candidates.
生物体液是通过不同的身体孔窍或透过皮肤渗透而释放出的含蛋白质的液体或悬液。这些体液是多种组织和细胞类型的产物,含有广泛、高度复杂且动态变化的蛋白质群体,既反映了起源细胞的转录程序,也记录了个体的健康状况。临床医生和研究人员很容易获取体液,因此蛋白质组学分析是临床研究、生育研究、口腔健康研究和法医调查的重要组成部分。当前的质谱(MS)数据集的动态范围高达六个数量级,并且与起源的组织类型一样多样。质谱有潜力在广泛的应用中提供信息,包括对人类生物学和病理学的基础研究、蛋白质功能的生化分析、生物标志物的发现与检测,以及在法医调查中,调查人员通过解读蛋白质谱来确定生物体液的身体部位来源。以下方法描述了一种样本处理工作流程,该流程在生物体液类型、样本状态(例如,从证据中提取的干燥样本或纯液体)和降解程度方面具有灵活性。这里描述的方法既适用于高灵敏度鸟枪法液相色谱 - 质谱联用(LC/MS)分析,也适用于基于质谱的生物标志物候选物的靶向(定性或定量)分析。