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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Qi, Dawei Zhang, Xiangmin Deng, Chunhui Yao, Guoping |
| Description | Country affiliation: China Author Affiliation: Yao G ( Department of Chemistry, Fudan University, Shanghai 200433, China.) |
| Abstract | In this study, magnetic carbonaceous (MC) microspheres prepared with a large-scale synthesis approach were developed as the novel substrate for enzyme immobilization, and the trypsin-immobilized MC microspheres were successfully applied to protein fast digestion. Firstly, MC microspheres with small size, strong magnetism, and biological compatibility were prepared through two-step solvothermal reactions. Secondly, MC microsphere surface was modified by 3-glycidoxypropyltrimethoxysilane (GLYMO). Finally, the enzyme was immobilized on the GLYMO-functionalized MC microspheres. The enzyme-immobilized magnetic microspheres were applied for fast protein digestion with microwave-assistance. Bovine serum albumin, myoglobin and cytochrome c, were used as model proteins to verify the digestion efficiency, and the digestion products were then characterized using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) with sequence coverage of 43%, 90% and 77%, respectively. The enzyme-immobilized magnetic particles were also successfully applied to the analysis of human pituitary extract. After database search, 485 proteins (p<0.01) were identified when the extract was digested by the microspheres. This opens a route for its future application in bottom-up proteomic analysis. |
| ISSN | 00219673 |
| Issue Number | 1-2 |
| Volume Number | 1215 |
| e-ISSN | 18733778 |
| Journal | Journal of Chromatography A |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2008-12-26 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Proteins Chemistry Trypsin Amino Acid Sequence Chromatography, Liquid Humans Hydrolysis Microscopy, Electron, Transmission Microspheres Microwaves Molecular Sequence Data Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry Journal Article Research Support, Non-u.s. Gov't Discipline Analytical Chemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Organic Chemistry Medicine Analytical Chemistry Biochemistry |
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