Proteomic analysis with integrated multiple dimensional liquid chromatography/mass spectrometry based on elution of ion exchange column using pH steps

	Proteomic analysis with integrated multiple dimensional liquid chromatography/mass spectrometry based on elution of ion exchange column using pH steps
	Dai, J; Shieh, CH; Sheng, QH; Zhou, H; Zeng, R
刊名	ANALYTICAL CHEMISTRY
	2005
卷号	77 期号:18 页码:5793-5799
通讯作者	Zeng, R (reprint author), Chinese Acad Sci, Res Ctr Proteome Anal, Key Lab Prote, Inst Biochem & Cell Biol,Shanghai Inst Biol Sci, 320 Yueyang Rd, Shanghai 200031, Peoples R China.,zr@sibs.ac.cn
英文摘要	A novel integrated multidimensional liquid chromatography (IMDL) method is demonstrated for the separation of peptide mixtures by two-dimensional HPLC coupled with ion trap mass spectrometry. The method uses an integrated column, containing both strong cation exchange and reversed-phase sections for two-dimensional liquid chromatography. The peptide mixture was fractionated by a pH step using a series of pH buffers, followed by reversed-phase chromatography. Since no salt was used during separation, the integrated multidimensional liquid chromatography can be directly connected to mass spectrometry for peptide analysis. The pH buffers were injected from an autosampler, and the entire process can be carried out on a one-dimensional liquid chromatography system. In a single analysis, the IMDL system, coupled with linear ion trap mass spectrometry, identified more than 2000 proteins in mouse liver. The peptides were eluted according to their pi distribution. The resolution of the pH fractionation is similar to 0.5 pH unit. The method has low overlapping across pH fractions, good resolution of peptide mixture, and good correlation of peptide pIs with pH steps. This method provides a technique for large-scale protein identification using existing one-dimensional HPLC systems.
学科主题	Chemistry
类目[WOS]	Chemistry, Analytical
关键词[WOS]	TANDEM MASS-SPECTROMETRY ; PROTEIN IDENTIFICATION ; YEAST PROTEOME ; SYSTEM
收录类别	SCI
语种	英语
WOS记录号	WOS:000231919400003
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1981]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Dai, J,Shieh, CH,Sheng, QH,et al. Proteomic analysis with integrated multiple dimensional liquid chromatography/mass spectrometry based on elution of ion exchange column using pH steps[J]. ANALYTICAL CHEMISTRY,2005,77(18):5793-5799.
APA	Dai, J,Shieh, CH,Sheng, QH,Zhou, H,&Zeng, R.(2005).Proteomic analysis with integrated multiple dimensional liquid chromatography/mass spectrometry based on elution of ion exchange column using pH steps.ANALYTICAL CHEMISTRY,77(18),5793-5799.
MLA	Dai, J,et al."Proteomic analysis with integrated multiple dimensional liquid chromatography/mass spectrometry based on elution of ion exchange column using pH steps".ANALYTICAL CHEMISTRY 77.18(2005):5793-5799.