Epitope mapping of 7S cashew antigen in complex with antibody by solution-phase H/D exchange monitored by FT-ICR mass spectrometry

J Mass Spectrom. 2015 Jun;50(6):812-9. doi: 10.1002/jms.3589.

Abstract

The potential epitope of a recombinant food allergen protein, cashew Ana o 1, reactive to monoclonal antibody, mAb 2G4, has been mapped by solution-phase amide backbone H/D exchange (HDX) monitored by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Purified mAb 2G4 was incubated with recombinant Ana o 1 (rAna o 1) to form antigen:monoclonal antibody (Ag:mAb) complexes. Complexed and uncomplexed (free) rAna o 1 were then subjected to HDX-MS analysis. Five regions protected from H/D exchange upon mAb binding are identified as potential conformational epitope-contributing segments.

Keywords: FTMS; allergen; hydrogen/deuterium exchange; protein complex.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Allergens / chemistry*
  • Allergens / immunology
  • Amino Acid Sequence
  • Anacardium / chemistry*
  • Deuterium Exchange Measurement / methods*
  • Epitope Mapping / methods*
  • Epitopes / chemistry*
  • Epitopes / immunology
  • Mass Spectrometry / methods
  • Models, Molecular
  • Molecular Sequence Data
  • Seed Storage Proteins / chemistry*
  • Seed Storage Proteins / immunology

Substances

  • Allergens
  • Epitopes
  • Seed Storage Proteins