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

Jingyueh Jeng, Mon Fang Lin, Fong Yu Cheng, Chen Sheng Yeh, Jentaie Shiea

研究成果: Article同行評審

30 引文 斯高帕斯(Scopus)

摘要

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.

原文English
頁(從 - 到)3060-3068
頁數9
期刊Rapid Communications in Mass Spectrometry
21
發行號18
DOIs
出版狀態Published - 2007

All Science Journal Classification (ASJC) codes

  • 分析化學
  • 光譜
  • 有機化學

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