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题名: Two-dimensional liquid chromatography separation of peptides using reversed-phase/weak cation-exchange mixed-mode column in first dimension
作者: Cai, Xiaoming1;  Guo, Zhimou1;  Xue, Xingya1;  Xu, Junyan1;  Zhang, Xiuli1;  Liang, Xinmiao1
关键词: Two-dimensional liquid chromatography ;  Mixed-mode chromatography ;  Polar-copolymerized ;  Proteomics ;  Peptide separation ;  LC/MS
刊名: JOURNAL OF CHROMATOGRAPHY A
发表日期: 2012-03-09
DOI: 10.1016/j.chroma.2011.06.042
卷: 1228, 页:242-249
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: Selection of a first-dimensional separation method is very important in proteomic studies performed with two-dimensional liquid chromatography (2D-LC), since reversed-phase chromatography (RPLC) is usually chosen as the second-dimensional separation method in most studies. In this paper, we assess the potential use of mixed-mode reversed-phase/weak cation-exchange (RP/WCX) chromatography in 2D-LC proteomic studies. First, a new RP/WCX mixed-mode stationary phase (named C18WCX) was synthesized based on the polar-copolymerized approach, and a C18WCX column with separation efficiency comparative to conventional C18 columns was developed. This new mixed-mode column primarily provides hydrophobic interactions under acidic condition, but can offer hydrophobic and cation-exchange interactions under neutral and weak basic conditions. An off-line 2D-RP/WCX-RPLC system was established using the C18WCX column with a mobile phase of pH 6.5 in the first dimension and a 08 column with a mobile phase of pH 3.0 in the second dimension. The orthogonality of this two-dimensional system, evaluated through the separation of 123 tryptic peptides, was shown to be higher than that of the conventional 2D-RP-RPLC approach. Applying this 2D-LC method to rat brain samples, we identified 1031 proteins and 4397 unique peptides. In addition, this new two-dimensional method improved the identification of basic peptides. Therefore, we propose that this novel 2D-RP/WCX-RPLC system can be used as an alternative approach for the two-dimensional separation of peptides. (C) 2011 Elsevier B.V. All rights reserved.
关键词[WOS]: PROTEIN IDENTIFICATION TECHNOLOGY ;  TANDEM MASS-SPECTROMETRY ;  SHOTGUN PROTEOMICS ;  STATIONARY-PHASE ;  MONOLITHIC COLUMNS ;  HORIZONTAL POLYMERIZATION ;  YEAST PROTEOME ;  HILIC-RP ;  SILICA ;  ENRICHMENT
语种: 英语
WOS记录号: WOS:000301406100024
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/138026
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Key Lab Separat Sci Analyt Chem, Dalian Inst Chem Phys, Dalian 116023, Peoples R China

Recommended Citation:
Cai, Xiaoming,Guo, Zhimou,Xue, Xingya,et al. Two-dimensional liquid chromatography separation of peptides using reversed-phase/weak cation-exchange mixed-mode column in first dimension[J]. JOURNAL OF CHROMATOGRAPHY A,2012,1228:242-249.
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