High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor
文献类型：期刊论文
| 作者 | Wu, Shuaibin1,2; Sun, Liangliang1,2; Ma, Junfeng1,2; Yang, Kaiguang1; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1 |
| 刊名 | talanta
 |
| 出版日期 | 2011-02-15 |
| 卷号 | 83期号:5页码:1748-1753 |
| 关键词 | Immobilized enzyme reactor Poly (acrylamide-co-methylenebisacrylamide) monolith Protein digestion Protein identification Proteomics |
| ISSN号 | 待补充 |
| 产权排序 | 1,1 |
| 通讯作者 | 张丽华 |
| 中文摘要 | high throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor |
| 英文摘要 | a poly (acrylamide-co-methylenebisacrylamide) (poly (aam-co-mba)) monolith was prepared by thermal polymerization in the 100 or 250 mu m i.d. capillary. the monolithic support was activated by ethylenediamine followed by glutaraldehyde. trypsin was then introduced to form an immobilized enzyme reactor (imer). the prepared imer showed a reliable mechanical stability and permeability (permeability constant k = 2.65 x 10(-13) m(2)). with bsa as the model protein, efficient digestion was completed within 20 s, yielding the sequence coverage of 57%, better than that obtained from the traditional in-solution digestion (42%), which took about 12 h. moreover. bsa down to femtomole was efficiently digested by the imer and positively identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (maldi-tof ms). to test the applicability of imer for complex sample profiling, proteins extracted from escherichia coli were digested by the imer and further analyzed by nanoreversed phase liquid chromatography-electrospray ionization-mass spectrometry (nanorplc-esi-ms/ms). in comparison to in-solution digestion, despite slightly fewer proteins were positively identified at a false discovery rate (fdr) of similar to 1% (333 vs 411), the digestion time used was largely shortened (20 s vs 24 h), implying superior digestion performance for the high throughput analysis of complex samples. (c) 2011 elsevier b.v. all rights reserved. |
| 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] | enabling protein digestion ; mass-spectrometry ; capillary electrochromatography ; peptide separation ; trypsin reactor ; identification ; proteomics ; system ; online |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000287110100065 |
| 公开日期 | 2012-07-09 |
| 源URL | [http://159.226.238.44/handle/321008/115315]  |
| 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, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wu, Shuaibin,Sun, Liangliang,Ma, Junfeng,et al. High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor[J]. talanta,2011,83(5):1748-1753. |
| APA | Wu, Shuaibin.,Sun, Liangliang.,Ma, Junfeng.,Yang, Kaiguang.,Liang, Zhen.,...&Zhang, Yukui.(2011).High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor.talanta,83(5),1748-1753. |
| MLA | Wu, Shuaibin,et al."High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor".talanta 83.5(2011):1748-1753. |
入库方式： OAI收割
来源：大连化学物理研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。