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Rapid and efficient protein digestion using trypsin-coated magnetic nanoparticles under pressure cycles
Cited 31 time in
Web of Science
Cited 35 time in Scopus
- Authors
- Issue Date
- 2011-01
- Publisher
- John Wiley & Sons Ltd.
- Citation
- Proteomics, Vol.11 No.2, pp.309-318
- 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 degrees 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 (similar to 1 min) and efficient digestions with more reproducible digestion results.
- ISSN
- 1615-9853
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