Abstract
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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Allergens / chemistry
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Allergens / genetics
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Amino Acid Sequence
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Anacardium / chemistry
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Anacardium / genetics
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Anacardium / immunology
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Animals
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Antibody Specificity
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Antigen-Antibody Complex / chemistry
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Antigens, Plant / chemistry*
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Antigens, Plant / genetics
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Cyclotrons
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Deuterium Exchange Measurement / methods*
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Epitope Mapping / methods*
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Epitopes / chemistry*
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Epitopes / genetics
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Fourier Analysis
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Goats
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Models, Molecular
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Molecular Sequence Data
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Peptide Fragments / chemistry
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Peptide Fragments / genetics
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Protein Subunits
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Spectrometry, Mass, Electrospray Ionization / methods*
Substances
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Allergens
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Ana o 2 allergen, cashew
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Antigen-Antibody Complex
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Antigens, Plant
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Epitopes
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Peptide Fragments
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Protein Subunits