Rapid protein identification using a microscale electrospray LC/MS system on an ion trap mass spectrometer
Rapid protein identification using a microscale electrospray LC/MS system on an ion trap mass spectrometer
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DOI：
10.1016/s1044-0305(97)00282-1
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发表时间：
1998-03-01
影响因子：
3.2
通讯作者：
Lee, TD
中科院分区：
文献类型：
--
作者：
Davis, MT;Lee, TD
发展了一种快速鉴定凝胶分离蛋白的方法。用胰酶消化凝胶蛋白后，用在线液相色谱、电喷雾电喷雾质谱(LC/MS)对多肽混合物进行分析。然后，将包含准确质量值或序列特定片段离子信息的质谱图数据与已知蛋白质序列的数据库进行匹配。LC/MS系统的主要特点是使用新型集成的微型LC柱-电喷雾界面和可变流量的溶剂输送，以优化样品加载和梯度洗脱的效率。有了这些改进，每个样品的分析只需要10分钟。该方法对取样量从50到500 fmol是常规的。为了跟上快速洗脱的LC峰的步伐，离子陷阱质谱仪的分析参数必须仔细调整。尽管专为快速LC分离而设计，集成柱-电喷雾界面也能够使用一种称为“峰停车”的技术对选定的组分进行扩展分析。(C)1998年美国质谱学学会。
A methodology has been developed for the rapid identification of gel separated proteins. Following in gel protein digestion with trypsin, the resulting peptide mixture is analyzed by on-line liquid chromatography, electrospray mass spectrometry (LC/MS). The mass spectral data containing either accurate mass values or sequence specific fragment ion information is then matched to a database of known protein sequences. Key features of the LC/MS system are the use of a novel integrated, microscale LC column-electrospray interface and variable flow solvent delivery to optimize the efficiency of sample loading and gradient elution. With these enhancements, only 10 min is required to analyze each sample. The method is routine for sample amounts ranging from 50 to 500 fmol. The analysis parameters for the ion trap mass spectrometer have to be carefully adjusted in order to keep pace with the rapidly eluting LC peaks. Although designed for rapid LC separations, the integrated column-electrospray interface is also able to provide extended analyses of selected components using a technique known as "peak parking." (C) 1998 American Society for Mass Spectrometry.