Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics
文献类型：期刊论文
| 作者 | Wu, Shuaibin1,3; Zhang, Lei; Yang, Kaiguang1; Liang, Zhen1; Zhang, Lihua; Zhang, Yukui1 |
| 刊名 | analytical and bioanalytical chemistry
 |
| 出版日期 | 2012 |
| 卷号 | 402期号:2页码:703-710 |
| 关键词 | Metal-ion chelation Polyacrylamide monolith Polyethylenimine Tryptic digestion Regenerable immobilized enzyme reactor |
| 产权排序 | 1,1 |
| 通讯作者 | 杨开广 |
| 英文摘要 | initially, a poly (glycidyl methacrylate-co-acrylamide-co-methylenebisacrylamide) monolith was prepared in the 100 mu m i.d. capillary, and then was grafted with polyethylenimine (mw, similar to 25,000) for adsorbing cu(2+), followed by chelating trypsin. as a result, efficient digestion for bsa (100 ng/mu l) was completed within 50 s via such immobilized enzyme reactor (imer); yielding 47% sequence coverage by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (maldi-tof ms) analysis. compared with the conventional method for preparing the metal-ion chelated imer, the regeneration of such imer can be achieved facilely by the respective 30 min desorption and re-adsorption of trypsin, and 51% sequence coverage was obtained for 50 s bsa digestion after regeneration. bsa down to femtomole was also efficiently digested by the prepared regenerable imer. meanwhile, after the consecutive digestion of myoglobin and bsa, there was not any mutual interference for both during maldi-tof ms identification, indicating the low nonspecific adsorption of such regenerable imer. to test the applicability of regenerable imer for complex sample profiling, proteins (150 ng) extracted from escherichia coli were digested within 80 s by the regenerable imer and further analyzed by nanoreversed phase liquid chromatography-electrospray ionization-mass spectrometry successfully, showing its practicability for the high throughput analysis of complex samples. |
| 学科主题 | 物理化学 |
| 类目[WOS] | biochemical research methods ; chemistry, analytical |
| 研究领域[WOS] | biochemistry & molecular biology ; chemistry |
| 关键词[WOS] | laser-desorption/ionization-time ; magnetic silica microspheres ; phase liquid-chromatography ; efficient protein digestion ; tandem mass-spectrometry ; trypsin immobilization ; identification ; online ; microreactor ; adsorption |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000298645300012 |
| 公开日期 | 2013-10-11 |
| 源URL | [http://159.226.238.44/handle/321008/118276]  |
| 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 R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China 2.Chinese Acad Sci, Dalian Inst Chem Phys, State Key Lab Catalysis, Dalian 116023, Peoples R China 3.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wu, Shuaibin,Zhang, Lei,Yang, Kaiguang,et al. Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics[J]. analytical and bioanalytical chemistry,2012,402(2):703-710. |
| APA | Wu, Shuaibin,Zhang, Lei,Yang, Kaiguang,Liang, Zhen,Zhang, Lihua,&Zhang, Yukui.(2012).Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics.analytical and bioanalytical chemistry,402(2),703-710. |
| MLA | Wu, Shuaibin,et al."Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics".analytical and bioanalytical chemistry 402.2(2012):703-710. |
入库方式： OAI收割
来源：大连化学物理研究所
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