填充型纳流液相色谱长柱的蛋白质组学分析性能考察  

Performance of Long Packed Nanoflow Liquid Chromatographic Columns in Proteomics Analysis

作　　者：王晓飞 秦飞 郭睿 张博[1] WANG Xiao-Fei;QIN Fei;GUO Rui;ZHANG Bo(College of Chemistry & Chemical Engineering,Xiamen University,Xiamen 361005,China;Xiamen Medical College,Xiamen 361023,China;Column Scientific Ltd,Xiamen 361102,China)

机构地区：[1]厦门大学化学化工学院,厦门361005 [2]厦门医学院,厦门361023 [3]厦门宜柱科技有限公司,厦门361102

出　　处：《分析化学》2019年第3期388-393,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(Nos.21475110;21535007;J1310024);中国博士后基金项目(No.2018M642572);福建省中青年教师教育科研项目(No.JT180645);厦门市科技计划项目(No.350Z20173019);中央高校基本科研业务费(No.20720160051)资助~~

摘　　要：纳流液相色谱长柱用于蛋白质组学分析中,可以显著提高多肽的分离度和检出率,进而提高蛋白质的鉴定数目和可信度。本研究基于纳流液相色谱-电喷雾质谱联用平台,考察了柱床长度为1.5 m的超长纳流液相色谱柱,将其用于蛋白质组学分析中。与常规纳流色谱柱相比,使用长柱时,复杂多肽样品得到了优良的分离效果。蛋白质组学数据检索结果表明,使用1.5 m长柱进行前端的色谱分离,采用质谱识别肽段数提升了83.6%,肽段数目提升了74.4%,唯一肽段数目提升了75.0%,蛋白质鉴定数目提升了38.4%。研究结果表明,这类填充型纳流液相色谱长柱为高分辨蛋白质组学提供了一种新的分离工具。Long nanoLC columns have been used in proteomics to improve the resolution and detectability of peptides to enhance protein identification number and analysis confidence.In this study,based on a nanoflow liquid chromatography-electrospray ionization mass spectrometry platform,the performance of a 1.5-m long packed nanoLC column and its applicability in proteomics were investigated.Compared with the commonly used short length nanoLC column,the long packed column achieved excellent resolution for the complex peptide mixture.The proteomics data showed that,using the 1.5-m long nanoLC column,the numbers of peptide spectrum matches,the identified peptides,the unique peptides and the protein groups were increased by 83.6%,74.4%,75.0%and 38.4%,respectively.The results demonstrated that the packed long nanoLC columns could be used as a new separation tool for high-resolution proteomics.

关 键 词：纳流液相色谱 毛细管液相色谱 填充柱 蛋白质组学 质谱 

分 类 号：Q51[生物学—生物化学] O652.63[理学—分析化学]