An integrated sample pretreatment platform for quantitative N-glycoproteome analysis with combination of on-line glycopeptide enrichment, deglycosylation and dimethyl labeling
文献类型：期刊论文
| 作者 | Weng, Yejing1,2; Qu, Yanyan1,2; Jiang, Hao1,2; Wu, Qi1,2; Zhang, Lihua1; Yuan, Huiming1; Zhou, Yuan1,2; Zhang, Xiaodan1; Zhang, Yukui1 |
| 刊名 | analytica chimica acta
 |
| 出版日期 | 2014-06-23 |
| 卷号 | 833页码:1-8 |
| 关键词 | Relative quantification N-glycoproteomes Integrated platform N-2-assisted interface Tumor metastasis |
| 英文摘要 | relative quantification of n-glycoproteomes shows great promise for the discovery of candidate biomarkers and therapeutic targets. the traditional protocol for quantitative analysis of glycoproteomes is usually off-line performed, and suffers from long sample preparation time, and the risk of sample loss or contamination due to manual manipulation. in this study, a novel integrated sample preparation platform for quantitative n-glycoproteome analysis was established, with combination of online n-glycopeptide capture by a hilic column, sample buffer exchange by a n-2-assisted hilic-rplc interface, deglycosylation by a hydrophilic pngase f immobilized enzymatic reactor (himer) and solid dimethyl labeling on a c18 precolumn. to evaluate the performance of such a platform, two equal aliquots of immunoglobulin g (igg) digests were sequentially pretreated, followed by maldi-tof ms analysis. the signal intensity ratio of heavy/light (h/l) labeled deglycosylated peptides with the equal aliquots was 1.00 (rsd = 6.2%, n = 3), much better than those obtained by the offline protocol, with h/l ratio as 0.76 (rsd = 11.6%, n = 3). additionally, the total on-line sample preparation time was greatly shortened to 160 min, much faster than that of offline approach (24 h). furthermore, such an integrated pretreatment platform was successfully applied to analyze the two kinds of hepatocarcinoma ascites syngeneic cell lines with high (hca-f) and low (hca-p) lymph node metastasis rates. for h/l labeled hca-p lysates with the equal aliquots, 99.6% of log2 ratios (h/l) of quantified glycopeptides ranged from -1 to 1, demonstrating high accuracy of the developed sample preparation strategy. by triplicated analysis of glycopeptides and non-glycopeptides of hca-f and hca-p lysates, 43 up-regulated and 30 down-regulated (hca-f/p)n-glycosylation sites, and 11 significantly changed n-glycoproteins were successfully quantified, and most of them were related to tumorigenesis and tumor metastasis. all these results demonstrate the developed integrated n-glycoprotein pretreatment platform is of great power for the accurate, precise and high-throughput analysis of n-glycoproteomes. (c) 2014 elsevier b.v. all rights reserved. |
| WOS标题词 | science & technology ; physical sciences |
| 类目[WOS] | chemistry, analytical |
| 研究领域[WOS] | chemistry |
| 关键词[WOS] | hydrophilic-interaction chromatography ; mass-spectrometry ; liquid-chromatography ; cancer-cells ; proteome analysis ; carcinoma cells ; glycosylation ; identification ; metastasis ; separation |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000337878800001 |
| 公开日期 | 2016-05-09 |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/145713]  |
| 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, Natl Chromatog Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100039, Peoples R China |
| 推荐引用方式 GB/T 7714 | Weng, Yejing,Qu, Yanyan,Jiang, Hao,et al. An integrated sample pretreatment platform for quantitative N-glycoproteome analysis with combination of on-line glycopeptide enrichment, deglycosylation and dimethyl labeling[J]. analytica chimica acta,2014,833:1-8. |
| APA | Weng, Yejing.,Qu, Yanyan.,Jiang, Hao.,Wu, Qi.,Zhang, Lihua.,...&Zhang, Yukui.(2014).An integrated sample pretreatment platform for quantitative N-glycoproteome analysis with combination of on-line glycopeptide enrichment, deglycosylation and dimethyl labeling.analytica chimica acta,833,1-8. |
| MLA | Weng, Yejing,et al."An integrated sample pretreatment platform for quantitative N-glycoproteome analysis with combination of on-line glycopeptide enrichment, deglycosylation and dimethyl labeling".analytica chimica acta 833(2014):1-8. |
入库方式： OAI收割
来源：大连化学物理研究所
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