Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis
文献类型：期刊论文
| 作者 | Zhu, Jun; Wang, Fangjun; Chen, Rui; Cheng, Kai; Xu, Bo; Guo, Zhimou; Liang, Xinmiao; Ye, Mingliang; Zou, Hanfa |
| 刊名 | analytical chemistry
 |
| 出版日期 | 2012-06-05 |
| 卷号 | 84期号:11页码:5146-5153 |
| ISSN号 | 0003-2700 |
| 产权排序 | 1,1 |
| 通讯作者 | 邹汉法 ; 叶明亮 |
| 英文摘要 | sample handling procedures including protein digestion, glycopeptide enrichment, and deglycosylation have significant impact on the performance of glycoproteome analysis. several glycoproteomic analysis systems were developed to integrate some of these sample preparation procedures. however, no microsystem integrates all of above three procedures together. in this work, we developed a glycoproteomic microreactor enabling seamless integration of all these procedures. in this reactor, trypsin digestion was accelerated by adding acetonitrile to 80%, and after acidification of protein digest by trifluoroacetic acid (tfa), the following hydrophilic interaction chromatography (hilic) enrichment and deglycosylation were sequentially performed without any desalting, lyophilization, or buffer exchange steps. the total processing time could be as short as 1.5 h. the detection limit of human igg as low as 30 fmol was also achieved. when applied to human serum glycoproteome analysis, a total number of 92, 178, and 221 unique n-glycosylation sites were identified from three replicate analyses of 10 nl, 100 nl, and 1 mu l of human serum, respectively. it was demonstrated that the glycoproteomic microreactor based method had very high sensitivity and was well suited for glycoproteome analysis of minute protein samples. |
| WOS标题词 | science & technology ; physical sciences |
| 学科主题 | 物理化学 |
| 类目[WOS] | chemistry, analytical |
| 研究领域[WOS] | chemistry |
| 关键词[WOS] | lectin affinity-chromatography ; phase liquid-chromatography ; mass-spectrometry ; proteome analysis ; glycosylation heterogeneity ; aberrant glycosylation ; maldi-ms ; glycopeptides ; plasma ; identification |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000304783100073 |
| 公开日期 | 2013-10-11 |
| 源URL | [http://159.226.238.44/handle/321008/117706]  |
| 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 | Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhu, Jun,Wang, Fangjun,Chen, Rui,et al. Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis[J]. analytical chemistry,2012,84(11):5146-5153. |
| APA | Zhu, Jun.,Wang, Fangjun.,Chen, Rui.,Cheng, Kai.,Xu, Bo.,...&Zou, Hanfa.(2012).Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis.analytical chemistry,84(11),5146-5153. |
| MLA | Zhu, Jun,et al."Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis".analytical chemistry 84.11(2012):5146-5153. |
入库方式： OAI收割
来源：大连化学物理研究所
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