Callesen Anne K, Mohammed Shabaz, Bunkenborg Jakob, Kruse Torben A, Cold Søren, Mogensen Ole, Christensen Rene dePont, Vach Werner, Jørgensen Per E, Jensen Ole N
Protein Research Group, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Rapid Commun Mass Spectrom. 2005;19(12):1578-86. doi: 10.1002/rcm.1960.
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 alpha-cyano-4-hydroxycinnamic acid with miniaturized SPE and MALDI-MS. Functionalized membrane discs with hydrophobic, ion-exchange or chelating properties allowed reproducible MALDI mass spectra (m/z 1000-12,000) 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.
通过基质辅助激光解吸/电离质谱（MALDI-MS）进行血清分析有望成为癌症和其他人类疾病早期诊断的临床工具。样品制备是在MALDI-MS中获得可重复且分辨率良好信号的关键；这是将基于MALDI-MS的诊断方法转化为临床应用的先决条件。我们研究了多种MALDI基质以及几种用于血清蛋白浓缩和脱盐的小型化固相萃取（SPE）方法，目的是通过MALDI-MS生成可重复的高质量蛋白质谱。我们开发了一种简单的血清分析方案，该方案将2,5-二羟基苯甲酸和α-氰基-4-羟基肉桂酸的基质混合物与小型化SPE和MALDI-MS相结合。具有疏水、离子交换或螯合特性的功能化膜盘能够从血清中获得可重复的MALDI质谱（m/z 1000 - 12,000）。在原理验证应用中，使用螯合材料的SPE和MALDI-MS鉴定出血清中的蛋白质峰，这些峰先前已被报道可用于区分乳腺癌患者和对照。这些初步结果表明，这种用于血清分析的简单SPE/MALDI-MS方法为临床蛋白质组学提供了一个通用且可扩展的平台。
