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题名: Integrated platform with a combination of online digestion and O-18 labeling for proteome quantification via an immobilized trypsin microreactor
作者: Zhang, Shen1, 2;  Yuan, Huiming1;  Zhao, Baofeng1;  Zhou, Yuan1;  Jiang, Hao1, 2;  Zhang, Lihua1;  Liang, Zhen1;  Zhang, Yukui1
刊名: ANALYST
发表日期: 2015
DOI: 10.1039/c5an00887e
卷: 140, 期:15, 页:5227-5234
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: A novel automated integrated platform for quantitative proteome analysis was established with a combination of online digestion of proteins and in situ O-18 labeling by an immobilized enzyme reactor (IMER); digests were captured and desalted by a C18 trap column, and peptides were analyzed by nanoRPLC-ESI-MS/MS. Bovine serum albumin (BSA) was used to evaluate the performance of the developed platform. Compared with traditional offline methods, not only the digestion and labeling time was shortened from 36 h to just 1 h, but also the labeling efficiency was improved from 95% to 99%. Furthermore, the back-exchange from O-18 to O-16 could also be efficiently avoided by the use of IMER. The platform was further evaluated by the quantitative analysis of 100 ng O-18 and O-16 online labeled yeast sample with a mixing ratio of 1 : 1, and the results showed significantly improved sensitivity and reproducibility, as well as improved quantitative accuracy than offline method. With these advantages, the integrated platform was finally applied to the quantitative profiling of 100 ng proteins extracted from two mouse hepato-carcinoma ascites syngeneic cell lines with high and low lymph node metastases rates, and ten differentially expressed proteins were successfully found, most of which were related to tumorigenesis and tumor metastasis. All these results demonstrate that the developed integrated platform can provide a new way for high efficiency O-18 labeling and the quantitative analysis of trace amounts of sample with high accuracy and high reproducibility.
关键词[WOS]: SPECTROMETRY-BASED PROTEOMICS ;  MASS-SPECTROMETRY ;  QUANTITATIVE PROTEOMICS ;  BACK-EXCHANGE ;  PRETREATMENT ;  THROUGHPUT ;  STRATEGIES ;  PEPTIDES ;  CELLS ;  MODEL
语种: 英语
WOS记录号: WOS:000357810800039
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/146384
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China

Recommended Citation:
Zhang, Shen,Yuan, Huiming,Zhao, Baofeng,et al. Integrated platform with a combination of online digestion and O-18 labeling for proteome quantification via an immobilized trypsin microreactor[J]. ANALYST,2015,140(15):5227-5234.
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