Lasaosa Maria, Jakoby Thomas, Delmotte Nathanaël, Huber Christian G, Heinzle Elmar, Tholey Andreas
Technische Biochemie, Universität des Saarlandes, Saarbrücken, Germany.
Proteomics. 2009 Oct;9(19):4577-81. doi: 10.1002/pmic.200900274.
A simple and fast screening method for the selection of fractions of first dimension separation to be analyzed in second dimension-MS/MS experiments in offline multidimensional liquid chromatographic separation schemes for shotgun proteome analysis was developed. The method is based on the measurement of total peptide content of the first dimension fractions by MALDI MS and was established using a tryptic digest of a bacterial proteome. The results of the screening process were in good agreement with those obtained in a detailed proteome analysis performed by RPxion-pair RP-MALDI TOF/TOF MS analysis. The method supports a straightforward planning of experiments, also enabling a reduction of overall measurement time in shotgun proteome analysis.
开发了一种简单快速的筛选方法，用于在鸟枪法蛋白质组分析的离线多维液相色谱分离方案中，选择在二维-MS/MS实验中进行分析的一维分离级分。该方法基于通过基质辅助激光解吸电离质谱（MALDI MS）测量一维级分的总肽含量，并使用细菌蛋白质组的胰蛋白酶消化物进行建立。筛选过程的结果与通过反相离子对反相-基质辅助激光解吸电离飞行时间/飞行时间质谱（RPxion-pair RP-MALDI TOF/TOF MS）分析进行的详细蛋白质组分析所获得的结果高度一致。该方法支持实验的直接规划，还能减少鸟枪法蛋白质组分析中的总体测量时间。
