High-throughput peptide mass mapping using a microdevice containing trypsin immobilized on a porous polymer monolith coupled to MALDI TOF and ESI TOF mass spectrometers

Dominic S. Peterson, Thomas Rohr, Frantisek Svec, Jean Frechet*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

142 Scopus citations

Abstract

An enzymatic microreactor with a volume of 470 nL has been prepared by immobilizing trypsin on a 10 cm long reactive porous polymer monolith located in a 100 μm i.d. fused silica capillary. This reactor affords suitable degrees of digestion of proteins even after very short residence times of less than 1 min. The performance is demonstrated with the digestion of eight proteins ranging in molecular mass from 2848 to 77 754. The digests were analyzed using mass spectrometry in two modes: off-line MALDI and in-line nanoelectrospray ionization. The large numbers of identified peptides enable a high degree of sequence coverage and positive identification of the proteins. The extent of sequence coverage decreases as the molecular mass of the digested protein increases.

Original languageEnglish (US)
Pages (from-to)563-568
Number of pages6
JournalJournal of Proteome Research
Volume1
Issue number6
DOIs
StatePublished - Nov 1 2002

Keywords

  • High-throughput protein identification
  • Immobilized enzyme
  • Mass spectrometry
  • Monolith
  • Proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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