Pepsin immobilization on an aldehyde-modified polymethacrylate monolith and its application for protein analysis

Wenjuan Han, Mika Yamauchi, Urara Hasegawa, Masanori Noda, Kiichi Fukui, André J. van der Vlies, Susumu Uchiyama, Hiroshi Uyama

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Polymer-based monoliths with interconnected porous structure have attracted much attention as a high-performance stationary phase for online digestion liquid chromatography-mass spectrometry (LC-MS) system. In this study, a poly(glycidyl methacrylate-. co-methyl methacrylate) (PGM) monolith prepared via thermally induced phase separation (TIPS) was used as a solid support to covalently immobilize pepsin. The PGM monolith was modified with aminoacetal to yield an aldehyde-bearing (PGM-CHO) monolith. Pepsin was immobilized onto the PGM-CHO monolith via reductive amination. The immobilized pepsin showed better pH and thermal stability compared with free pepsin. Furthermore, the PGM-CHO monolith modified with pepsin was applied for online protein digestion followed by LC-MS and LC-MS/MS analyses. As a result, a larger number of peptides are reproducibly identified compared to those by polystyrene/divinylbenzene particle (POROS)-based online pepsin column.

Original languageEnglish (US)
Pages (from-to)505-510
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume119
Issue number5
DOIs
StatePublished - May 1 2015

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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