Rapid and efficient protein digestion using trypsin-coated magnetic nanoparticles under pressure cycles

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

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)309-318
Number of pages10
JournalProteomics
Volume11
Issue number2
DOIs
Publication statusPublished - 2011 Jan 2

Keywords

  • Enzyme coatings
  • Magnetic nanoparticles
  • Nanoproteomics
  • Pressure cycling technology
  • Protein digestion

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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