Department of Medical and Clinical Biophysics, Faculty of Medicine, University of P.J. Šafárik in Košice, Košice, Slovakia.
Department of Clinical Pharmacology and Pharmacoepidemiology, Heidelberg University Hospital, Heidelberg, Germany.
Proteomics. 2021 May;21(9):e2000318. doi: 10.1002/pmic.202000318. Epub 2021 Mar 5.
Multiple applications of proteomics in life and health science, pathology and pharmacology, require handling size-limited cell and tissue samples. During proteomic sample preparation, analyte loss in these samples arises when standard procedures are used. Thus, specific considerations have to be taken into account for processing, that are summarised under the term microproteomics (μPs). Microproteomic workflows include: sampling (e.g., flow cytometry, laser capture microdissection), sample preparation (possible disruption of cells or tissue pieces via lysis, protein extraction, digestion in bottom-up approaches, and sample clean-up) and analysis (chromatographic or electrophoretic separation, mass spectrometric measurements and statistical/bioinformatic evaluation). All these steps must be optimised to reach wide protein dynamic ranges and high numbers of identifications. Under optimal conditions, sampling is adapted to the studied sample types and nature, sample preparation isolates and enriches the whole protein content, clean-up removes salts and other interferences such as detergents or chaotropes, and analysis identifies as many analytes as the instrumental throughput and sensitivity allow. In the suggested review, we present and discuss the current state in μP applications for processing of small number of cells (cell μPs) and microscopic tissue regions (tissue μPs).
蛋白质组学在生命科学和健康科学、病理学和药理学中的多种应用需要处理有限大小的细胞和组织样本。在蛋白质组学样品制备过程中，当使用标准程序时，这些样品中的分析物会丢失。因此，必须针对处理考虑具体因素，这些因素在术语“微量蛋白质组学 (μPs)”下进行了总结。微量蛋白质组学工作流程包括：采样（例如，流式细胞术、激光捕获显微切割）、样品制备（通过裂解、蛋白质提取、消化（自上而下的方法）和样品净化可能破坏细胞或组织碎片）以及分析（色谱或电泳分离、质谱测量和统计/生物信息学评估）。所有这些步骤都必须进行优化，以达到广泛的蛋白质动态范围和高鉴定数量。在最佳条件下，采样适用于研究的样本类型和性质，样品制备分离和富集整个蛋白质含量，净化去除盐和其他干扰物，如洗涤剂或变性剂，分析尽可能多地识别仪器的吞吐量和灵敏度允许的分析物。在建议的综述中，我们展示并讨论了用于处理少量细胞（细胞 μPs）和微观组织区域（组织 μPs）的微量蛋白质组学应用的现状。
