Integrated Platform for Proteome Analysis with Combination of Protein and Peptide Separation via Online Digestion

	Integrated Platform for Proteome Analysis with Combination of Protein and Peptide Separation via Online Digestion
	Yuan, Huiming 1,2; Zhang, Lihua 1; Hou, Chunyan 1,2; Zhu, Guijie 1,2; Tao, Dingyin 1,2; Liang, Zhen 1; Zhang, Yukui 1
刊名	analytical chemistry
	2009-11-01
卷号	81 期号:21 页码:8708-8714
通讯作者	张丽华
产权排序	1;1
英文摘要	an integrated platform with the combination of protein and peptide separation was established via online protein digestion, by which proteins were first separated by a microcolumn packed with mixed weak anion and weak cation exchange (wax/wcx) particles under a series of salt steps, online digested by a trypsin immobilized microenzymatic reactor (imer), trapped and desalted by two parallel cs precolumns, separated by microreversed-phase liquid chromatography (mu rplc) under a linear gradient of organic modifier concentration, and finally identified by electrospray ionization-ms/ms (esi-ms/ms). to evaluate the performance of such a platform, a mixture of myoglobin, cytochrome c, bovine serum albumin (bsa), and alpha-casein, with mass ranging from 25 ng to 2 mu g, was analyzed. compared to the methods by offline protein fractionation and shotgun based strategy, the analysis time, including sample preparation, digestion, desalting, separation, and detection, was shortened from ca. 30 to 5 h, and cytochrome c with abundance of 25 ng could be identified with improved sequence coverage. furthermore, such an integrated platform was successfully applied into the analysis of proteins extracted from human lung cancer cells. compared with the results obtained by the shotgun approach, the identified protein number was increased by 30%. all these results demonstrated that such an integrated approach would be an attractive alternative to commonly applied approaches for proteome research.
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, analytical
研究领域[WOS]	chemistry
关键词[WOS]	multidimensional liquid-chromatography ; tandem mass-spectrometry ; top-down proteomics ; ion-exchange ; identification ; microreactor ; bottom ; system ; column ; monoliths
收录类别	SCI
原文出处	false
语种	英语
WOS记录号	WOS:000276191900013
公开日期	2010-11-30
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/102393]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Yuan, Huiming,Zhang, Lihua,Hou, Chunyan,et al. Integrated Platform for Proteome Analysis with Combination of Protein and Peptide Separation via Online Digestion[J]. analytical chemistry,2009,81(21):8708-8714.
APA	Yuan, Huiming.,Zhang, Lihua.,Hou, Chunyan.,Zhu, Guijie.,Tao, Dingyin.,...&Zhang, Yukui.(2009).Integrated Platform for Proteome Analysis with Combination of Protein and Peptide Separation via Online Digestion.analytical chemistry,81(21),8708-8714.
MLA	Yuan, Huiming,et al."Integrated Platform for Proteome Analysis with Combination of Protein and Peptide Separation via Online Digestion".analytical chemistry 81.21(2009):8708-8714.