A facile microdialysis interface for on-line desalting and identification of proteins by nano-electrospray ionization mass spectrometry
A facile microdialysis interface for on-line desalting and identification of proteins by nano-electrospray ionization mass spectrometry
复制标题
一种简便的微透析接口，用于通过纳米电喷雾电离质谱法在线脱盐和鉴定蛋白质
DOI：
10.1002/rcm.3622
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发表时间：
2008-08-01
影响因子：
2
通讯作者：
Zhang, Yukui
中科院分区：
文献类型：
--
作者：
Sun, Liangliang;Duan, Jicheng;Zhang, Yukui
盐，特别是无机盐对电喷雾电离质谱（ESI-MS）的影响是限制其应用的最大障碍之一。在众多的透析方法中，微透析技术对大分子，如蛋白质是有利的。在这项工作中，采用醋酸纤维素中空纤维膜，（MWCO 3000 Da），构建了一个简单、方便、有效的微透析界面，死体积小于1 μ L，用于高盐缓冲液中蛋白质的在线质谱分析。该系统已成功地应用于肌红蛋白在高浓度醋酸铵或氯化钠缓冲溶液中的分析。实验结果表明，通过使用这样的微透析接口，盐浓度，即使高达1 M，可以降低至少2个数量级，而样品损失小于10%，展示了这样的接口在扩大应用的潜力nano-ESI-MS在大分子分析。版权所有（c）2008约翰威利父子有限公司。
The adverse effect of salts, especially inorganic salts, on electrospray ionization mass spectrometry (ESI-MS) is one of the most serious obstacles that might limit its application. Among the numerous desalting approaches, the microdialysis technique is favorable for large molecules, such as proteins. In this work, employing a hollow fiber membrane of cellulose acetate (MWCO 3000 Da), a simple, facile and efficient microdialysis interface with the dead volume of less than 1 mu L was constructed for the on-line desalting and identification of proteins dissolved in high salt concentration buffer by nano-ESI-MS. Furthermore, with counterflow added, the desalting procedure was accelerated, and could be finished within 1 min. This system was successfully applied to the analysis of myoglobin dissolved in either high concentration ammonium acetate or sodium chloride buffer. The experimental results showed that, by using such a microdialysis interface, the salt concentration, even as high as 1 M, could be decreased by at least 2 orders of magnitude, while sample loss was less than 10%, demonstrating the potential of such an interface in broadening the application of nano-ESI-MS in the analysis of large molecules. Copyright (c) 2008 John Wiley & Sons, Ltd.