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Two-dimensional liquid chromatography separation of peptides using reversed-phase/weak cation-exchange mixed-mode column in first dimension
Cai, Xiaoming; Guo, Zhimou; Xue, Xingya; Xu, Junyan; Zhang, Xiuli; Liang, Xinmiao
KeywordTwo-dimensional Liquid Chromatography Mixed-mode Chromatography Polar-copolymerized Proteomics Peptide Separation Lc/ms
Source PublicationJOURNAL OF CHROMATOGRAPHY A
2012-03-09
DOI10.1016/j.chroma.2011.06.042
Volume1228Pages:242-249
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordPROTEIN IDENTIFICATION TECHNOLOGY ; TANDEM MASS-SPECTROMETRY ; SHOTGUN PROTEOMICS ; STATIONARY-PHASE ; MONOLITHIC COLUMNS ; HORIZONTAL POLYMERIZATION ; YEAST PROTEOME ; HILIC-RP ; SILICA ; ENRICHMENT
AbstractSelection 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.
Language英语
WOS IDWOS:000301406100024
Citation statistics
Cited Times:33[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/138026
Collection中国科学院大连化学物理研究所
AffiliationChinese Acad Sci, Key Lab Separat Sci Analyt Chem, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
Recommended Citation
GB/T 7714
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.
APA Cai, Xiaoming,Guo, Zhimou,Xue, Xingya,Xu, Junyan,Zhang, Xiuli,&Liang, Xinmiao.(2012).Two-dimensional liquid chromatography separation of peptides using reversed-phase/weak cation-exchange mixed-mode column in first dimension.JOURNAL OF CHROMATOGRAPHY A,1228,242-249.
MLA Cai, Xiaoming,et al."Two-dimensional liquid chromatography separation of peptides using reversed-phase/weak cation-exchange mixed-mode column in first dimension".JOURNAL OF CHROMATOGRAPHY A 1228(2012):242-249.
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