Enrichment of serum low-molecular-weight proteins using C-18 absorbent   under urea/dithiothreitol denatured environment

doi:10.1016/j.ab.2009.10.047

CORC > 北京大学 > 生命科学学院

	Enrichment of serum low-molecular-weight proteins using C-18 absorbent under urea/dithiothreitol denatured environment
	Wu, Jing ; An, Yuan ; Pu, Hai ; Shan, Yue ; Ren, Xiaoqing ; An, Mingrui ; Wang, Qingsong ; Wei, Shicheng ; Ji, Jianguo
刊名	分析生物化学
	2010
关键词	Serum Low-molecular-weight protein Enrichment C-18 absorbent Proteomics PROTEOMIC ANALYSIS MASS-SPECTROMETRY REMOVE ALBUMIN HUMAN PLASMA GEL ELECTROPHORESIS ULTRAFILTRATION SCATTERING DEPLETION SAMPLES
DOI	10.1016/j.ab.2009.10.047
英文摘要	Serum low-molecular-weight proteins (LMWPs, molecular weight <30 kDa) are closely related to the body physiological and pathological situations, whereas many difficulties are encountered when enriching and fractionating them. Using C-18 absorbent (100 A) enrichment and fractionation under urea/dithiothreitol (DTT) denatured environment followed by 60% acetonitrile (ACN) elution, serum LMWPs could be enriched more than 100-fold and were evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SIDS-PAGE), two-dimensional gel electrophoresis (2-DE), and isotope-coded affinity tag (ICAT) labeling quantification. Proteins existing in human serum at low nanograms/milliliter (ng/ml) levels, such as myeloid-related proteins (MRPs), could be identified directly from 2-DE coupled with matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF/TOF MS) and LTQ-Orbitrap MS. Sixteen proteins were confidentially identified and quantified using ICAT labeling and liquid chromatography-tandem mass spectrometry (LC-MS/MS). By virtue of its easy operation and high reproducibility to process large quantity complex serum samples, this method has potential uses in enriching LMWPs either in serum or in cell and tissue samples. (C) 2009 Elsevier Inc. All rights reserved.; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000273956500005&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical; SCI(E); PubMed; 7; ARTICLE; 1; 34-44; 398
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/162559]
专题	生命科学学院
推荐引用方式 GB/T 7714	Wu, Jing,An, Yuan,Pu, Hai,et al. Enrichment of serum low-molecular-weight proteins using C-18 absorbent under urea/dithiothreitol denatured environment[J]. 分析生物化学,2010.
APA	Wu, Jing.,An, Yuan.,Pu, Hai.,Shan, Yue.,Ren, Xiaoqing.,...&Ji, Jianguo.(2010).Enrichment of serum low-molecular-weight proteins using C-18 absorbent under urea/dithiothreitol denatured environment.分析生物化学.
MLA	Wu, Jing,et al."Enrichment of serum low-molecular-weight proteins using C-18 absorbent under urea/dithiothreitol denatured environment".分析生物化学 (2010).