色谱

色谱

• 研究论文 • 上一篇    下一篇

基于整体材料的微流控芯片反相液相色谱-串联质谱平台用于蛋白质分析

梁玉1,2, 吴慈1,2, 戴忠鹏1, 梁作成1, 梁振1, 张丽华1*, 张玉奎1   

  1. 1. 中国科学院分离分析化学重点实验室, 大连化学物理研究所, 国家色谱研究分析中心, 辽宁 大连 116023; 2. 中国科学院研究生院, 北京 100039
  • 收稿日期:2011-04-08 修回日期:2011-05-07 出版日期:2011-06-28 发布日期:2011-07-25
  • 通讯作者: 张丽华,研究员,博士生导师,主要从事生物分子的高效分离与表征等相关研究工作. Tel: (0411)84379720
  • 基金资助:

    国家自然科学基金(20935004)、科技部基础研究计划(2007CB714503, 2007CB914100)和科技部科技支撑计划(2008BAK41B02, 2009BAK59B02)项目.

Microchip-based reversed-phase liquid chromatography-tandem mass spectrometry platform for protein analysis

LIANG Yu1,2, WU Ci1,2, DAI Zhongpeng1, LIANG Zuocheng1, LIANG Zhen1, ZHANG Lihua1*, ZHANG Yukui1   

  1. 1 Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, National Chromatographic R. & A. Center, Dalian 116023, China; 2. Graduate University of Chinese Academy of Sciences, Beijing 100039, China
  • Received:2011-04-08 Revised:2011-05-07 Online:2011-06-28 Published:2011-07-25

PDF

563

被引次数

5

摘要： 微流控芯片高效液相色谱-串联质谱系统具有高通量、高灵敏度等优点,已成为生物样品分析的热点领域之一。本文在玻璃芯片上以甲基丙烯酸十二酯(LMA)和三羟甲基丙烷三甲基丙烯酸酯(TMPTMA)为单体,制备了以聚丙烯酸酯整体材料为固定相的捕集柱和分离柱。通过在芯片通道末端连接细内径的毛细管作为芯片-质谱接口,并以常规的液相色谱泵和微阀控制流体,构建了芯片反相液相色谱-电喷雾串联质谱(RPLC-ESI-MS/MS)平台,并将其用于分析牛血清白蛋白(BSA)的酶解产物。经过3次平行分析,BSA的序列覆盖率分别为39.37%、37.89%和34.10%(相对标准偏差为7.3%)。采用不同批次制作的芯片构建RPLC-ESI-MS/MS平台,对BSA酶解产物进行分析,其序列覆盖率相当。上述结果表明,该平台具有灵敏度高和重现性好等优点,有望用于蛋白质样品的快速分离和高灵敏度鉴定。

关键词: 串联质谱, 蛋白质分析, 反相液相色谱, 微流控芯片, 整体材料

Abstract: Due to the high throughput and high sensitivity, the hyphenation of microchip-based high performance liquid chromatography with tandem mass spectrometry has been paid much attention. In our recent work, with poly(lauryl methacrylate-co-trimethylolpropane trimethacrylate) monolithic materials prepared in microchannels as trap and separation columns, conventional micro-liquid chromatography pumps and valves for fluidic control, and a small-bore open-tube capillary attached to the outlet channel as chip-mass spectrometer (MS) interface, the microchip-based reversed-phase liquid chromatography-tandem mass spectrometry (RPLC-MS/MS) platform was established, and applied for the identification of proteins. By such platform, 100 ng digest of bovine serum albumin (BSA) was successfully analyzed with the sequence coverages as 39.37%, 37.89% and 34.10% (with the relative standard deviation (RSD) of 7.3%) in three runs, separately. To evaluate the chip-to-chip reproducibility, BSA was identified by such platform with the microchips from different batches containing trap column, separation column and chip-MS interface. The obtained sequence coverage and the number of peptides identified were comparable. All these results showed high sensitivity and good reproducibility of such platform, demonstrating the great potential for rapid protein analysis.

Key words: monolith, protein analysis, tandem mass spectrometry (MS/MS), microchip, reversed-phase liquid chromatography (RPLC)