Rapid and efficient protein digestion using trypsin-coated magnetic nanoparticles under pressure cycles.
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| Abstract |
Trypsin-coated magnetic nanoparticles (EC-TR/NPs), prepared via a simple multilayer random crosslinking of the trypsin molecules onto magnetic nanoparticles, were highly stable and could be easily captured using a magnet after the digestion was complete. EC-TR/NPs showed a negligible loss of trypsin activity after multiple uses and continuous shaking, whereas the conventional immobilization of covalently attached trypsin on NPs resulted in a rapid inactivation under the same conditions due to the denaturation and autolysis of trypsin. A single model protein, a five-protein mixture, and a whole mouse brain proteome were digested at atmospheric pressure and 37°C for 12 h or in combination with pressure cycling technology at room temperature for 1 min. In all cases, EC-TR/NPs performed equally to or better than free trypsin in terms of both the identified peptide/protein number and the digestion reproducibility. In addition, the concomitant use of EC-TR/NPs and pressure cycling technology resulted in very rapid (∼1 min) and efficient digestions with more reproducible digestion results.
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| Authors | Byoungsoo Lee, Daniel Lopez-Ferrer, Byoung Chan Kim, Hyon Bin Na, Yong Il Park, Karl K Weitz, Marvin G Warner, Taeghwan Hyeon, Sang-Won Lee, Richard D Smith, Jungbae Kim |
| Journal | Proteomics (Proteomics) Vol. 11 Issue 2 Pg. 309-18 (Jan 2011) ISSN: 1615-9861 [Electronic] Germany |
| PMID | 21204257 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.) |
| Copyright | Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. |
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