Identification of mammalian cell lines using MALDI-TOF and LC-ESI-MS/MS mass spectrometry
Identification of mammalian cell lines using MALDI-TOF and LC-ESI-MS/MS mass spectrometry
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DOI：
10.1016/j.jasms.2005.12.007
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发表时间：
2006-04-01
影响因子：
3.2
通讯作者：
Smith, LM
中科院分区：
文献类型：
--
作者：
Zhang, X;Scalf, M;Smith, LM
复杂生物样品的直接质谱分析在蛋白质组学领域正成为一种越来越有用的技术。基质辅助激光解吸/电离质谱（MALDI-MS）是一种快速、灵敏的分析工具，非常适合从复杂样品中获得肽和蛋白质的分子量。本文描述了一种快速、简单的细胞蛋白分析方法，其中哺乳动物细胞直接在MALDI基质2,5-二羟基苯甲酸（DHB）中裂解，并使用MALDI飞行时间（TOF）进行质量分析。利用这些分析中产生的独特MALDI质谱“指纹”，可以区分几种不同的哺乳动物细胞系。MALDI-后源衰减（PSD）、MALDI串联飞行时间（TOF-TOF）、MALDI-傅立叶变换离子回旋共振（FTICR）、纳米流液相色谱-电喷雾电离和串联质谱（LC-ESI-MS/MS）等技术被用于鉴定MALDI光谱中所代表的蛋白质。对在DHB中裂解的细胞上清进行胰酶消化，随后进行LC-ESI-MS/MS分析，是迄今为止鉴定蛋白质最成功的方法。
Direct mass spectrometric analysis of complex biological samples is becoming an increasingly useful technique in the field of proteomics. Matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS) is a rapid and sensitive analytical tool well suited for obtaining molecular weights of peptides and proteins from complex samples. Here, a fast and simple approach to cellular protein profiling is described in which mammalian cells are lysed directly in the MALDI matrix 2,5-dihydroxybenzoic acid (DHB) and mass analyzed using MALDI-time of flight (TOF). Using the unique MALDI mass spectral "fingerprint" generated in these analyses, it is possible to differentiate among several different mammalian cell lines. A number of techniques, including MALDI-post source decay (PSD), MALDI tandem time-of-flight (TOF-TOF), MALDI-Fourier transform ion cyclotron resonance (FTICR), and nanoflow liquid chromatography followed by electrospray ionization and tandem mass spectrometry (LC-ESI-MS/MS) were employed to attempt to identify the proteins represented in the MALDI spectra. Performing a tryptic digestion of the supernatant of the cells lysed in DHB with subsequent LC-ESI-MS/MS analysis was by far the most successful method to identify proteins.