Lee, Byoungsoo Lopez-Ferrer, Daniel Kim, Byoung Chan Na, Hyon Bin Park, Yong Il Weitz, Karl K. Warner, Marvin G. Hyeon, Taeghwan Lee, Sang-Won Smith, Richard D. Kim, Jungbae 2024-01-20T18:00:58Z 2024-01-20T18:00:58Z 2021-09-05 2011-01 1615-9853 https://pubs.kist.re.kr/handle/201004/130757 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. English WILEY-BLACKWELL INTENSITY FOCUSED ULTRASOUND GAS-PHASE FRACTIONATION MASS-SPECTROMETRY QUANTITATIVE PROTEOMICS CYCLING TECHNOLOGY SAMPLE PREPARATION PEPTIDES THROUGHPUT SYSTEM MS Rapid and efficient protein digestion using trypsin-coated magnetic nanoparticles under pressure cycles Article 10.1002/pmic.201000378 1 PROTEOMICS, v.11, no.2, pp.309 - 318 PROTEOMICS 11 2 309 318 scie scopus 000285884400012 2-s2.0-78650789063 Biochemical Research Methods Biochemistry & Molecular Biology Biochemistry & Molecular Biology Article INTENSITY FOCUSED ULTRASOUND GAS-PHASE FRACTIONATION MASS-SPECTROMETRY QUANTITATIVE PROTEOMICS CYCLING TECHNOLOGY SAMPLE PREPARATION PEPTIDES THROUGHPUT SYSTEM MS Enzyme coatings Magnetic nanoparticles Nanoproteomics Pressure cycling technology Protein digestion