A standardized and reproducible urine preparation protocol for cancer biomarkers discovery.
A standardized and reproducible urine preparation protocol for cancer biomarkers discovery.
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DOI：
10.4137/bic.s17991
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Beretov J;Wasinger VC;Schwartz P;Graham PH;Li Y
一个合适和标准化的蛋白质纯化技术对于保持一致性和允许蛋白质组学研究之间的数据比较以发现尿液生物标记物是必不可少的。最终，应努力使尿液制备方案标准化。这项研究的目的是开发一种最佳的分析方案，以实现最大的蛋白质产量，并确保该方法适用于检测区分疾病和非疾病状态的尿蛋白模式。在这项初步研究中，我们比较了七种不同的尿样制备方法，以去除盐分，并沉淀和分离尿蛋白。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)图谱显示，丙酮和三氯乙酸(TCA)结合高速离心法(HSC)顺序制备的尿蛋白分离效果最好，且保留了最多的尿蛋白。因此，该方法是所有进一步尿蛋白分析的首选方法。
A suitable and standardized protein purification technique is essential to maintain consistency and to allow data comparison between proteomic studies for urine biomarker discovery. Ultimately, efforts should be made to standardize urine preparation protocols. The aim of this study was to develop an optimal analytical protocol to achieve maximal protein yield and to ensure that this method was applicable to examine urine protein patterns that distinguish disease and disease-free states. In this pilot study, we compared seven different urine sample preparation methods to remove salts, and to precipitate and isolate urinary proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles showed that the sequential preparation of urinary proteins by combining acetone and trichloroacetic acid (TCA) alongside high speed centrifugation (HSC) provided the best separation, and retained the most urinary proteins. Therefore, this approach is the preferred method for all further urine protein analysis.