文献类型：期刊文献

英文题名：Sequential enrichment of singly- and multiply-phosphorylated peptides with zwitterionic hydrophilic interaction chromatography material

作者：Sheng, Qianying[1];Yang, Kaiya[1];Xue, Xingya[3];Li, Xiuling[3,4];Guo, Zhimou[3];Shen, Aijin[3];Ke, Yanxiong[5];Lan, Minbo[1,2];Liang, Xinmiao

机构：[1]E China Univ Sci & Technol, Shanghai Key Lab Funct Mat Chem, Shanghai 200237, Peoples R China;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[3]Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China;[4]Nantong Univ, Coinnovat Ctr Neuroregenerat, Nantong 226019, Peoples R China;[5]E China Univ Sci & Technol, Engn Res Ctr Pharmaceut Proc Chem, Shanghai 200237, Peoples R China

年份：2015

卷号：1413

起止页码：47

外文期刊名：JOURNAL OF CHROMATOGRAPHY A

收录：;EI(收录号：20153501209064);WOS:【SCI-EXPANDED(收录号:WOS:000361577600005)】；

基金：This work was supported by National High Technology Research and Development Program of China (863 Program, 2012AA020203), National Science Foundation of China (21135005 and 21475129) and the Fundamental Research Funds for the Central Universites (222201514022).

语种：英文

外文关键词：Zwitterionic material; Mixed-mode retention mechanisms; Sequential enrichment; Multiply-phosphorylated peptides; Mass spectrometry

摘要：An interesting and novel method for the selective and sequential enrichment of singly- and multiply-phosphorylated peptides with a zwitterionic material "Click TE-Cys" is presented. Retention mechanisms between phosphopeptides and Click TE-Cys are systematically investigated by checking the influence of acetonitrile content, pH value, and buffer concentration on the retention of phosphopeptides. Both hydrophilic interaction and electrostatic interaction are involved in retention between phosphopeptides and Click TE-Cys. Based on these results, an optimized method is established for selective enrichment of phosphopeptides using Click TE-Cys. This method not only exhibits high selectivity for phosphopeptides, but also fractionates singly- and multiply-phosphorylated peptides into two fractions. This method was evaluated using relatively complex samples, including peptide mixtures of alpha-casein and bovine serum albumin (BSA) at a molar ratio of 1:10 and skim milk. This efficient and optimized protocol has great potential for enriching multiply-phosphorylated peptides and could be a valuable tool for specific enrichment of phosphopeptides in phosphoproteome analysis. (C) 2015 Elsevier B.V. All rights reserved.