On-line Immobilized Enzyme Microreactor Capillary Zone Electrophoresis-Mass Spectrometry for Peptide Mapping

Methods Mol Biol. 2022:2531:77-91. doi: 10.1007/978-1-0716-2493-7_6.

Abstract

Peptide mapping is a routine procedure for protein characterization in proteomics. This bottom-up analysis requires digestion of proteins into peptides before liquid chromatography- or capillary zone electrophoresis-mass spectrometry (LC-MS or CZE-MS, respectively). Proteins are usually digested off-line using proteolytic enzymes, typically trypsin, in solution or immobilized on appropriate supports. As an alternative, here we describe on-line immobilized enzyme microreactor capillary zone electrophoresis-mass spectrometry (IMER-CZE-MS) for a straightforward, rapid, and efficient protein digestion followed by separation, detection, and characterization of the generated peptides.

Keywords: Bottom-up proteomics; Capillary zone electrophoresis; Immobilized enzyme; In-line; Mass spectrometry; Microcartridge; Microreactor; On-line.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electrophoresis, Capillary* / methods
  • Enzymes, Immobilized* / chemistry
  • Mass Spectrometry
  • Peptide Mapping
  • Peptides / metabolism
  • Proteins
  • Trypsin / chemistry

Substances

  • Enzymes, Immobilized
  • Peptides
  • Proteins
  • Trypsin