Fiedler Georg Martin, Ceglarek Uta, Leichtle Alexander, Thiery Joachim
Institute of Laboratory Medicine, Clinical Chemistry and Molecular Diagnostics, University Hospital Leipzig, Medical Faculty, University Leipzig, Leipzig, Germany.
Methods Mol Biol. 2010;641:47-63. doi: 10.1007/978-1-60761-711-2_4.
Proteome/peptidome profiling of human urine is a promising tool for the discovery of novel disease-associated biomarkers. However, a wide range of preanalytic variables influence the results of proteome/peptidome analysis regardless of the method used. We present a validated pretreatment protocol, which allows standardization of preanalytic modalities and facilitates reproducible peptidome profiling of human urine by means of magnetic bead (MB) separation in combination with MALDI-TOF MS. Such a procedure is necessary for generating consistent and reliable data from which meaningful results may be obtained for biomarker discovery and general proteomic experiments.
对人类尿液进行蛋白质组/肽组分析是发现新型疾病相关生物标志物的一种很有前景的工具。然而,无论使用何种方法,一系列广泛的分析前变量都会影响蛋白质组/肽组分析的结果。我们提出了一种经过验证的预处理方案,该方案能够实现分析前模式的标准化,并通过磁珠(MB)分离结合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)促进对人类尿液进行可重复的肽组分析。这样的程序对于生成一致且可靠的数据是必要的,从中可以获得有意义的结果用于生物标志物发现和一般蛋白质组学实验。