An optimization of the LC-MS/MS workflow for deep proteome profiling on an Orbitrap Fusion
文献类型：期刊论文
| 作者 | Nie, Litong1,2; Zhu, Mingrui1,2; Sun, Shengnan1,2; Zhai, Linhui1,2; Wu, Zhixiang3; Qian, Lili1,2; Tan, Minjia1,2
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| 刊名 | ANALYTICAL METHODS
 |
| 出版日期 | 2016 |
| 卷号 | 8期号:2页码:425-434 |
| ISSN号 | 1759-9660 |
| DOI | 10.1039/c5ay01900a |
| 文献子类 | Article |
| 英文摘要 | The development of high-resolution mass spectrometers (MS) has greatly advanced the system-wide proteomic profiling and protein post-translational modification (PTM) studies. However, in contrast to current genomic sequencing technologies, huge time cost and laborious workload are the major bottlenecks of current MS-based proteomic approaches for large-scale in-depth proteome sequencing of biological samples. Here we present a stepwise optimization of MS parameters and an off-line reverse phase HPLC fractionation method in the first tribrid MS platform-Orbitrap Fusion, which integrates quadrupole, ultrahigh field Orbitrap and linear ion trap mass analyzers. With off-line high pH separation, we identified more than 5000 proteins using a regular short reverse phase C18 column (10 cm x 75 mu m, 3 mu m particle size) in a single one hour LC-MS run and 8493 proteins with 6 orders of magnitude of dynamic range in only 10 hour MS running time. Our study provided a fast, cost-efficient and amenable method for deep proteomic analysis and quantification of large-scale biological samples. Significantly, this strategy would facilitate the proteomic disease biomarker discovery. |
| WOS关键词 | SPECTROMETRY-BASED PROTEOMICS ; PEPTIDE IDENTIFICATION RATES ; MASS-SPECTROMETRY ; QUANTITATIVE PROTEOMICS ; SHOTGUN PROTEOMICS ; CELL-LINE ; QUANTIFICATION ; PERFORMANCE ; CANCER ; PHOSPHOPROTEOME |
| 资助项目 | Natural Science Foundation of China[31370814] ; National Science and Technology Major Project "Key New Drug Creation and Manufacturing Program" of China[2014ZX09507-002] ; National Basic Research Program of China (973 Program)[2014CBA02004] ; Shanghai Municipal Science and Technology Commission[14DZ2261100] ; Shanghai Municipal Science and Technology Commission[15410723100] |
| WOS研究方向 | Chemistry ; Food Science & Technology ; Spectroscopy |
| 语种 | 英语 |
| WOS记录号 | WOS:000367243600025 |
| 出版者 | ROYAL SOC CHEMISTRY |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/276273]  |
| 专题 | 化学蛋白质组学研究中心 中科院受体结构与功能重点实验室 新药研究国家重点实验室 |
| 通讯作者 | Tan, Minjia |
| 作者单位 | 1.Chinese Acad Sci, Shanghai Inst Mat Med, Chem Prote Ctr, Shanghai 200031, Peoples R China; 2.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, Shanghai 200031, Peoples R China; 3.Shanghai Jiao Tong Univ, Xinhua Hosp, Dept Pediat Surg, Shanghai 200030, Peoples R China |
| 推荐引用方式 GB/T 7714 | Nie, Litong,Zhu, Mingrui,Sun, Shengnan,et al. An optimization of the LC-MS/MS workflow for deep proteome profiling on an Orbitrap Fusion[J]. ANALYTICAL METHODS,2016,8(2):425-434. |
| APA | Nie, Litong.,Zhu, Mingrui.,Sun, Shengnan.,Zhai, Linhui.,Wu, Zhixiang.,...&Tan, Minjia.(2016).An optimization of the LC-MS/MS workflow for deep proteome profiling on an Orbitrap Fusion.ANALYTICAL METHODS,8(2),425-434. |
| MLA | Nie, Litong,et al."An optimization of the LC-MS/MS workflow for deep proteome profiling on an Orbitrap Fusion".ANALYTICAL METHODS 8.2(2016):425-434. |
入库方式： OAI收割
来源：上海药物研究所
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