Using high-concentration trypsin-immobilized magnetic nanoparticles for rapid in situ protein digestion at elevated temperature.
Rapid Commun Mass Spectrom
; 21(18): 3060-8, 2007.
Article
en En
| MEDLINE
| ID: mdl-17705254
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.
Buscar en Google
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Mapeo Peptídico
/
Tripsina
/
Proteínas
/
Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
/
Espectrometría de Masa por Ionización de Electrospray
/
Nanopartículas
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
/
Prognostic_studies
Idioma:
En
Revista:
Rapid Commun Mass Spectrom
Año:
2007
Tipo del documento:
Article
País de afiliación:
Taiwán