Rapid mass spectrometric conversion of tissue biopsy samples into permanent quantitative digital proteome maps.
Rapid mass spectrometric conversion of tissue biopsy samples into permanent quantitative digital proteome maps.
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每个临床标本都是固有的独特、有限和不可再生的。因此，诸如组织活检的小样本通常在有限数量的分析之后被完全消耗。在这里，我们提出了一种方法，使快速和可重复的少量组织（近似活检获得的数量）转换成一个单一的，永久的数字文件，代表质谱可测量的蛋白质组的样品。该方法结合了压力循环技术（PCT）和SWATH质谱（MS），并且可以分析，重新分析，比较和计算机挖掘所得的蛋白质组图谱，以检测和定量多个样品中的特定蛋白质。我们用这种方法处理和转换18个活检样品从9个肾细胞癌患者SWATH-MS碎片离子图。从这些蛋白质组图谱中，我们检测并定量了2,000多种蛋白质，在所有样品中具有高度的重现性。所鉴定的蛋白质清楚地将肿瘤肾组织与健康组织分开，并区分不同的组织形态学肾癌亚型。
Clinical specimens are each inherently unique, limited and non-renewable. As such, small samples such as tissue biopsies are often completely consumed after a limited number of analyses. Here we present a method that enables fast and reproducible conversion of a small amount of tissue (approximating the quantity obtained by a biopsy) into a single, permanent digital file representing the mass spectrometry-measurable proteome of the sample. The method combines pressure cycling technology (PCT) and SWATH mass spectrometry (MS), and the resulting proteome maps can be analyzed, re-analyzed, compared and mined in silico to detect and quantify specific proteins across multiple samples. We used this method to process and convert 18 biopsy samples from 9 renal cell carcinoma patients into SWATH-MS fragment ion maps. From these proteome maps we detected and quantified more than 2,000 proteins with a high degree of reproducibility across all samples. The identified proteins clearly separated tumorous kidney tissues from healthy tissue, and differentiated distinct histomorphological kidney cancer subtypes.