We have developed a streamlined proteomic sample preparation protocol termed Accelerated Barocycler Lysis and Extraction (ABLE) that substantially reduces the time and cost of tissue sample processing. ABLE is based on pressure cycling technology (PCT) for rapid tissue solubilization and reliable, controlled proteolytic digestion. Here, a previously reported PCT based protocol was optimized using 1–4 mg biopsy punches from rat kidney. The tissue denaturant urea was substituted with a combination of sodium deoxycholate (SDC) and N-propanol. ABLE produced comparable numbers of protein identifications in half the sample preparation time, being ready for MS injection in 3 h compared with 6 h for the conventional urea based method. To validate ABLE, it was applied to a diverse range of rat tissues (kidney, lung, muscle, brain, testis), human HEK 293 cell lines, and human ovarian cancer samples, followed by SWATH-mass spectrometry (SWATH-MS). There were similar numbers of quantified proteins between ABLE-SWATH and the conventional method, with greater than 70% overlap for all sample types, except muscle (58%). The ABLE protocol offers a standardized, high-throughput, efficient, and reproducible proteomic preparation method that when coupled with SWATH-MS has the potential to accelerate proteomics analysis to achieve a clinically relevant turn-around time.

中文翻译：

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质谱加速临床蛋白质组学的Barocycler裂解和提取样品制备

我们已经开发了一种简化的蛋白质组学样品制备方案，称为“加速Barocycler裂解和提取（ABLE）”，可大大减少组织样品处理的时间和成本。ABLE基于压力循环技术（PCT），可实现快速组织溶解和可靠的，受控的蛋白水解消化。在这里，先前报道的基于PCT的方案使用了来自大鼠肾脏的1-4 mg活检穿孔器进行了优化。组织变性尿素被脱氧胆酸钠（SDC）和N的组合取代-丙醇。ABLE在一半的样品制备时间中产生了相当数量的蛋白质鉴定结果，可以在3小时内完成MS注入，而传统的基于尿素的方法需要6小时。为了验证ABLE，将其应用于多种大鼠组织（肾脏，肺，肌肉，脑，睾丸），人HEK 293细胞系和人卵巢癌样品，然后使用SWATH-质谱法（SWATH-MS）。在ABLE-SWATH和常规方法之间，定量蛋白质的数量相似，除肌肉（58％）外，所有样品类型的重叠率均超过70％。ABLE协议提供了一种标准化，高通量，高效且可重现的蛋白质组学制备方法，当与SWATH-MS结合使用时，它有可能加速蛋白质组学分析以实现与临床相关的周转时间。