Abstract Urine reflects the renal function and urinary and kidney systems, but it may also reflect the presence of cancer in other parts of the body. Urine also has potential for providing prognostic information during therapeutic treatments thanks to non-invasive monitoring. A quick and reproducible protein purification procedure is essential to allow data comparison between proteomic studies in urine biomarker discovery. The article describes a simple, reproducible and cheap sample preparation procedure with a maximum protein yield (400 µg) obtained from only 10 mL of urine utilising cut-off filter desalting and digestion. The reported procedure removes yellowish background coloration residues and thus prevents the errors in spectrophotometric protein concentration determination. Different extraction solvents used in the presented procedure point to the possibility of partial elimination of abundant proteins (albumin and keratin family), as well as to the improvement of the sequence coverage of proteins identified, which helps to reveal changes in the urinary proteome. With this workflow, proteins can be easily obtained on standard laboratory equipment within 3 h. Data are available via ProteomeXchange with identifier PXD019738.

中文翻译：

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鸟枪法蛋白质组学尿样制备的优化

摘要 尿液反映肾功能和泌尿和肾脏系统，但也可能反映身体其他部位是否存在癌症。由于无创监测，尿液还具有在治疗过程中提供预后信息的潜力。快速且可重复的蛋白质纯化程序对于尿液生物标志物发现中蛋白质组学研究之间的数据比较至关重要。该文章描述了一种简单、可重现且廉价的样品制备程序，利用截断过滤器脱盐和消解仅从 10 mL 尿液中获得最大蛋白质产量 (400 µg)。报告的程序去除了黄色背景着色残留物，从而防止了分光光度法蛋白质浓度测定中的错误。所提出的程序中使用的不同提取溶剂表明有可能部分消除丰富的蛋白质（白蛋白和角蛋白家族），以及提高所鉴定蛋白质的序列覆盖率，这有助于揭示尿蛋白质组的变化。使用此工作流程，可以在 3 小时内在标准实验室设备上轻松获得蛋白质。数据可通过 ProteomeXchange 获得，标识符为 PXD019738。