Serum protein profiling by miniaturized solid-phase extraction and matrix-assisted laser desorption/ionization mass spectrometry
Serum protein profiling by miniaturized solid-phase extraction and matrix-assisted laser desorption/ionization mass spectrometry
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DOI：
10.1002/rcm.1960
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发表时间：
2005-01-01
影响因子：
2
通讯作者：
Jensen, ON
中科院分区：
文献类型：
--
作者：
Callesen, AK;Mohammed, S;Jensen, ON
基质辅助激光解吸/电离质谱仪(MALDI-MS)作为一种早期诊断癌症和其他人类疾病的临床工具，具有广阔的应用前景。样品制备是在MALDI-MS中获得重现性和良好分辨率信号的关键，也是将基于MALDI-MS的诊断方法转化为临床应用的先决条件。我们研究了几种MALDI基质和几种用于血清蛋白质浓缩和脱盐的微型固相萃取(SPE)方法，目的是利用MALDI-MS建立可重复性的、高质量的蛋白质图谱。我们开发了一种简单的血清分析方法，将2，5-二羟基苯甲酸和x-氰基-4-羟基肉桂酸的基质混合物与微型固相萃取和MALDI-MS结合起来。具有疏水、离子交换或螯合特性的功能化膜盘允许从血清中获得可重现的MALDI质谱图(m/z1000-12000)。在一项原理验证应用中，使用螯合材料和MALDI-MS的固相萃取确定了血清中的蛋白质峰，这些蛋白质峰以前曾被报道用于区分乳腺癌诊断患者和对照组。这些初步结果表明，这种简单的血清SPE/MALDI-MS方法为临床蛋白质组学提供了一个通用和可扩展的平台。版权所有(C)2005 John Wiley&Sons，Ltd.
Serum profiling by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) holds promise as a clinical tool for early diagnosis of cancer and other human diseases. Sample preparation is key to achieving reproducible and well-resolved signals in MALDI-MS; a prerequisite for translation of MALDI-MS based diagnostic methods to clinical applications. We have investigated a number of MALDI matrices and several miniaturized solid-phase extraction (SPE) methods for serum protein concentration and desalting with the aim of generating reproducible, high-quality protein profiles by MALDI-MS. We developed a simple protocol for serum profiling that combines a matrix mixture of 2,5-dihydroxybenzoic acid and x-cyano-4-hydroxycinnarnic acid with miniaturized SPE and MALDI-MS. Functionalized membrane discs with hydrophobic, ion-exchange or chelating properties allowed reproducible MALDI mass spectra (m/z 1000-12000) to be obtained from serum. In a proof-of-principle application, SPE with chelating material and MALDI-MS identified protein peaks in serum that had been previously reported for distinguishing a person diagnosed with breast cancer from a control. These preliminary results indicate that this simple SPE/MALDI-MS method for serum profiling provides a versatile and scalable platform for clinical proteomics. Copyright (c) 2005 John Wiley & Sons, Ltd.