Trypsin immobilization in ordered porous polymer membranes for effective protein digestion
文献类型：期刊论文
| 作者 | Qiao, Juan1; Kim, Jin Yong2; Wang, Yuan Yuan1; Qi, Li1; Wang, Fu Yi1; Moon, Myeong Hee2 |
| 刊名 | ANALYTICA CHIMICA ACTA
 |
| 出版日期 | 2016-02-04 |
| 卷号 | 906页码:156-164 |
| 关键词 | Porous polymer membrane enzyme reactor Breath figure method On-line proteolysis |
| 英文摘要 | Fast and effective protein digestion is a vital process for mass spectrometry (MS) based protein analysis. This study introduces a porous polymer membrane enzyme reactor (PPMER) coupled to nanoflow liquid chromatography-tandem MS (nLC-ESI-MS/MS) for on-line digestion and analysis of proteins. Poly (styrene- co-maleic anhydride) (PS-co-MAn) was fabricated by the breath figure method to make a porous polymer membrane in which the MAn group was covalently bound to enzyme. Based on this strategy, microscale PPMER (mu PPMER) was constructed for on-line connection with the nLC-ESI-MS/MS system. Its capability for enzymatic digestion with bovine serum albumin (BSA) was evaluated with varied digestion periods. The on-line proteolysis of BSA and subsequent analysis with mu PPMER-nLC-ESI-MS/MS revealed that peptide sequence coverage increased from 10.3% (digestion time 10 min) to 89.1% (digestion time 30 min). mu PPMER can efficiently digest proteins due to the microscopic confinement effect, showing its potential application in fast protein identification and protease immobilization. Applications of on-line digestion using mu PPMER with human plasma and urinary proteome samples showed that the developed on-line method yielded equivalent or better performance in protein coverage and identified more membrane proteins than the in-solution method. This may be due to easy accommodation of hydrophobic membrane proteins within membrane pores. (C) 2015 Elsevier B.V. All rights reserved. |
| 收录类别 | SCI |
| 语种 | 英语 |
| 公开日期 | 2016-05-09 |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/29724]  |
| 专题 | 化学研究所_活体分析化学实验室 |
| 作者单位 | 1.Chinese Acad Sci, Beijing Natl Lab Mol Sci, Key Lab Analyt Chem Living Biosyst, Inst Chem, Beijing 100190, Peoples R China 2.Yonsei Univ, Dept Chem, Seoul 120749, South Korea |
| 推荐引用方式 GB/T 7714 | Qiao, Juan,Kim, Jin Yong,Wang, Yuan Yuan,et al. Trypsin immobilization in ordered porous polymer membranes for effective protein digestion[J]. ANALYTICA CHIMICA ACTA,2016,906:156-164. |
| APA | Qiao, Juan,Kim, Jin Yong,Wang, Yuan Yuan,Qi, Li,Wang, Fu Yi,&Moon, Myeong Hee.(2016).Trypsin immobilization in ordered porous polymer membranes for effective protein digestion.ANALYTICA CHIMICA ACTA,906,156-164. |
| MLA | Qiao, Juan,et al."Trypsin immobilization in ordered porous polymer membranes for effective protein digestion".ANALYTICA CHIMICA ACTA 906(2016):156-164. |
入库方式： OAI收割
来源：化学研究所
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