Using high‐concentration trypsin‐immobilized magnetic nanoparticles for rapid in situ protein digestion at elevated temperature

Citations Over TimeTop 18% of 2007 papers

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 microg/microL) at 57 degrees 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.

Related Papers

• → Improvement of in‐gel digestion protocol for peptide mass fingerprinting by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry(2001)237 cited
• → Peptide-Mass Profiles of Polyvinylidene Difluoride-Bound Proteins by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry in the Presence of Nonionic Detergents(1996)112 cited
• → Improvement of an in-gel tryptic digestion method for matrix-assisted laser desorption/ionization-time of flight mass spectrometry peptide mapping by use of volatile solubilizing agents(2001)36 cited
• → Laser desorption/ionization—Mass spectrometry using mesoporous silicate as matrix for the analysis of various molecules(2007)12 cited
• → Improvement of an in-gel tryptic digestion method for matrix-assisted laser desorption/ionization-time of flight mass spectrometry peptide mapping by use of volatile solubilizing agents(2001)2 cited