Journal
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
Volume 24, Issue 7, Pages 1016-1025Publisher
AMER CHEMICAL SOC
DOI: 10.1007/s13361-013-0644-7
Keywords
Hydrogen/deuterium exchange; Allergen; Cashew; Epitope mapping; FTMS
Funding
- NIH [R01 GM78359]
- NSF Division of Materials Research [DMR-06-54118]
- USDA NIFA-AFRI [2010-04213]
- Almond Board of California
- State of Florida
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The potential epitopes of a recombinant food allergen protein, cashew Ana o 2, reactive to polyclonal antibodies, were mapped by solution-phase amide backbone H/D exchange (HDX) coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Ana o 2 polyclonal antibodies were purified in the serum from a goat immunized with cashew nut extract. Antibodies were incubated with recombinant Ana o 2 (rAna o 2) to form antigen:polyclonal antibody (Ag:pAb) complexes. Complexed and uncomplexed (free) rAna o 2 were then subjected to HDX-MS analysis. Four regions protected from H/D exchange upon pAb binding are identified as potential epitopes and mapped onto a homologous model.
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