A fully automated 2-D LC-MS method utilizing online continuous pH and RP gradients for global proteome analysis
文献类型：期刊论文
| 作者 | Zhou, H; Dai, J; Sheng, QH; Li, RX; Shieh, CH; Guttman, A; Zeng, R |
| 刊名 | ELECTROPHORESIS
 |
| 出版日期 | 2007 |
| 卷号 | 28期号:23页码:4311-4319 |
| 关键词 | continuous gradient MS pH gradient strong cation exchange 2-D LC |
| 通讯作者 | Zeng, R (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Syst Biol, 320 Yueyang Rd, Shanghai 200031, Peoples R China.,zr@sibs.ac.cn |
| 英文摘要 | The conventional 2-D LC-MS/MS setup for global proteome analysis was based on online and offline salt gradients (step and continuous) using strong-cation-exchange chromatography in conjunction with RP chromatography and MS. The use of the online system with step salt elution had the possibility of resulting in peptide overlapping across fractions. The offline mode had the option to operate with continuous salt gradient to decrease peak overlap, but exhibited decreased robustness, lower reproducibility, and sample loss during the process. Due to the extensive washing requirement between the chromatography steps, online continuous gradient was not an option for salt elution. In this report, a fully automated, online, and continuous gradient (pH continuous online gradient, pCOG) 2-D LC-MS/MS system is introduced that provided excellent separation and identification power. The pH gradient-based elution provided more basic peptides than that of salt-based elution. Fraction overlap was significantly minimized by combining pH and continuous gradient elutions. This latter approach also increased sequence coverage and the concomitant confidence level in protein identification. The salt and pH elution-based 2-D LC-MS/MS approaches were compared by analyzing the mouse liver proteome. |
| 学科主题 | Biochemistry & Molecular Biology; Chemistry |
| 类目[WOS] | Biochemical Research Methods ; Chemistry, Analytical |
| 关键词[WOS] | SPECTROMETRY-BASED PROTEOMICS ; TANDEM MASS-SPECTROMETRY ; MULTIDIMENSIONAL LIQUID-CHROMATOGRAPHY ; YEAST PROTEOME ; CELL-CULTURE ; PROTEINS ; IDENTIFICATION ; OPTIMIZATION ; SEPARATIONS ; COMPLEXES |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000252037300008 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1630]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Zhou, H,Dai, J,Sheng, QH,et al. A fully automated 2-D LC-MS method utilizing online continuous pH and RP gradients for global proteome analysis[J]. ELECTROPHORESIS,2007,28(23):4311-4319. |
| APA | Zhou, H.,Dai, J.,Sheng, QH.,Li, RX.,Shieh, CH.,...&Zeng, R.(2007).A fully automated 2-D LC-MS method utilizing online continuous pH and RP gradients for global proteome analysis.ELECTROPHORESIS,28(23),4311-4319. |
| MLA | Zhou, H,et al."A fully automated 2-D LC-MS method utilizing online continuous pH and RP gradients for global proteome analysis".ELECTROPHORESIS 28.23(2007):4311-4319. |
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