imFASP: An integrated approach combining in-situ filter-aided sample pretreatment with microwave-assisted protein digestion for fast and efficient proteome sample preparation

doi:10.1016/j.aca.2016.01.049

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	imFASP: An integrated approach combining in-situ filter-aided sample pretreatment with microwave-assisted protein digestion for fast and efficient proteome sample preparation
	Zhao, Qun 1; Fang, Fei 1,2; Wu, Ci 1,2; Wu, Qi 1,2; Liang, Yu 1; Liang, Zhen 1; Zhang, Lihua 1; Zhang, Yukui 1
刊名	ANALYTICA CHIMICA ACTA
	2016-03-17
卷号	912 页码:58-64
关键词	In-situ Sample preparation Filter Microwave Trace proteome analysis
ISSN号	0003-2670
DOI	10.1016/j.aca.2016.01.049
通讯作者	Zhang, Lihua(lihuazhang@dicp.ac.cn)
英文摘要	An integrated sample preparation method, termed "imFASP", which combined in-situ filter-aided sample pretreatment and microwave-assisted trypsin digestion, was developed for preparation of microgram and even nanogram amounts of complex protein samples with high efficiency in 1 h. For imFASP method, proteins dissolved in 8 M urea were loaded onto a filter device with molecular weight cut off (MWCO) as 10 kDa, followed by in-situ protein preconcentration, denaturation, reduction, alkylation, and microwave-assisted tryptic digestion. Compared with traditional in-solution sample preparation method, imFASP method generated more protein and peptide identifications (IDs) from preparation of 45 mg Escherichia coli protein sample due to the higher efficiency, and the sample preparation throughput was significantly improved by 14 times (1 h vs. 15 h). More importantly, when the starting amounts of E. coli cell lysate decreased to nanogram level (50-500 ng), the protein and peptide identified by imFASP method were improved at least 30% and 44%, compared with traditional in-solution preparation method, suggesting dramatically higher peptide recovery of imFASP method for trace amounts of complex proteome samples. All these results demonstrate that the imFASP method developed here is of high potential for high efficient and high throughput preparation of trace amounts of complex proteome samples. (C) 2016 Elsevier B.V. All rights reserved.
资助项目	National Basic Research Program of China[2012CB910604] ; National Natural Science Foundation[21405153] ; National Natural Science Foundation[21505136] ; Creative Research Group Project by NSFC[21321064]
WOS研究方向	Chemistry
语种	英语
出版者	ELSEVIER SCIENCE BV
WOS记录号	WOS:000370680000007
资助机构	National Basic Research Program of China ; National Natural Science Foundation ; Creative Research Group Project by NSFC
内容类型	期刊论文
源URL	[http://ir.imr.ac.cn/handle/321006/126953]
专题	金属研究所_中国科学院金属研究所
通讯作者	Zhang, Lihua
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, 457 Zhongshan Rd, Dalian 116023, Peoples R China 2.Grad Sch Chinese Acad Sci, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Zhao, Qun,Fang, Fei,Wu, Ci,et al. imFASP: An integrated approach combining in-situ filter-aided sample pretreatment with microwave-assisted protein digestion for fast and efficient proteome sample preparation[J]. ANALYTICA CHIMICA ACTA,2016,912:58-64.
APA	Zhao, Qun.,Fang, Fei.,Wu, Ci.,Wu, Qi.,Liang, Yu.,...&Zhang, Yukui.(2016).imFASP: An integrated approach combining in-situ filter-aided sample pretreatment with microwave-assisted protein digestion for fast and efficient proteome sample preparation.ANALYTICA CHIMICA ACTA,912,58-64.
MLA	Zhao, Qun,et al."imFASP: An integrated approach combining in-situ filter-aided sample pretreatment with microwave-assisted protein digestion for fast and efficient proteome sample preparation".ANALYTICA CHIMICA ACTA 912(2016):58-64.