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  

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

作　　者：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] 

机构地区：[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

出　　处：《色谱》2011年第6期469-474,共6页Chinese Journal of Chromatography

基　　金：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

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。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 micsochips 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.

关 键 词：微流控芯片 反相液相色谱 串联质谱 整体材料 蛋白质分析 

分 类 号：O658[理学—分析化学]