Qiu Feng, Liu Hong-Ying, Zhang Xue-Ji, Tian Ya-Ping
Department of Clinical Biochemistry, Chinese PLA General Hospital, 28 Fu-Xing Road, Beijing 100853, China.
Front Biosci (Landmark Ed). 2009 Jan 1;14(10):3712-23. doi: 10.2741/3482.
MALDI-TOF MS is being widely applied to analyze serum samples for the diagnosis of human diseases and for the identification of potential biomarkers. However, different types of magnetic beads afford different purification capability for low abundance proteins and peptides in body fluids. The aim of our study was to establish a proteome fractionation technique for choosing magnetic beads for proteomic analysis in breast cancer research. We used three different kinds of magnetic beads (MB-HIC C8, MB-IMAC-Cu and MB-WCX) to pre-analyze 28 breast cancer patients and 24 healthy control serum samples for proteomic profiling by MALDI-TOF MS analysis. The serum samples purified by MB-WCX group provided the best proteomic pattern .These samples had the most average peak numbers, the highest peak intensities, and the best capturing ability of low abundance proteins or peptides in serum samples. Therefore, we chose MB-WCX as our best magnetic beads to pre-analyze large-scale cancer and healthy control samples. The application of the standardized, pre-analytical serum samples with right type of magnetic bead-based fractionation will decrease variability of proteome patterns in human serum assessed by MALDI-TOF MS.
基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)正被广泛应用于分析血清样本,以用于人类疾病的诊断和潜在生物标志物的鉴定。然而,不同类型的磁珠对体液中低丰度蛋白质和肽段具有不同的纯化能力。我们研究的目的是建立一种蛋白质组分级分离技术,用于在乳腺癌研究中选择用于蛋白质组分析的磁珠。我们使用三种不同类型的磁珠(MB-HIC C8、MB-IMAC-Cu和MB-WCX)对28例乳腺癌患者和24例健康对照者的血清样本进行预分析,通过MALDI-TOF MS分析进行蛋白质组谱分析。MB-WCX组纯化的血清样本提供了最佳的蛋白质组图谱。这些样本具有最多的平均峰数、最高的峰强度,以及对血清样本中低丰度蛋白质或肽段的最佳捕获能力。因此,我们选择MB-WCX作为最佳磁珠,用于对大规模癌症和健康对照样本进行预分析。应用标准化的、基于正确类型磁珠分级分离的分析前血清样本,将降低通过MALDI-TOF MS评估的人血清蛋白质组图谱的变异性。