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Using high‐concentration trypsin‐immobilized magnetic nanoparticles for rapid in situ protein digestion at elevated temperature
Author(s) -
Jeng Jingyueh,
Lin MonFang,
Cheng FongYu,
Yeh ChenSheng,
Shiea Jentaie
Publication year - 2007
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3191
Subject(s) - chemistry , digestion (alchemy) , trypsin , chromatography , lysozyme , peptide , mass spectrometry , matrix assisted laser desorption/ionization , desorption , sample preparation in mass spectrometry , biochemistry , enzyme , electrospray ionization , adsorption , organic chemistry
Abstract We describe an innovative approach – using a high concentration of trypsin‐modified magnetic nanoparticles (TMNPs) – for the rapid and efficient digestion of proteins at elevated temperature. The required digestion time could be reduced to less than 10 s. After digestion, the TMNPs were collected magnetically from the sample solution for reuse and the digested peptides were characterized using matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF) mass spectrometry. Protein digestion was optimized when using the TMNPs (5 µg/µL) at 57°C; a significantly high peptide coverage was achieved for protein identification (e.g., 98% for lysozyme). Although a high concentration of TMNPs was used for digestion, the short digestion time led to much lower amounts of trypsin peptides being produced through self‐digestion. As a result, interference in the mass spectrometric detection of the peptide ions was reduced significantly. Copyright © 2007 John Wiley & Sons, Ltd.