title: Rapid screening for potential epitopes reactive with a polycolonal antibody by solution-phase H/D exchange monitored by FT-ICR mass spectrometry.
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.
authors.affiliation: Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL 32306, USA.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.forename: Kyle A
authors.initials: KA
authors.lastname: Noble
authors.forename: Yuan
authors.initials: Y
authors.lastname: Mao
authors.forename: Nicolas L
authors.initials: NL
authors.lastname: Young
authors.forename: Shridhar K
authors.initials: SK
authors.lastname: Sathe
authors.forename: Kenneth H
authors.initials: KH
authors.lastname: Roux
authors.forename: Alan G
authors.initials: AG
authors.lastname: Marshall
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000235:genetics; D000595:Amino Acid Sequence; D031021:Anacardium / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000818:Animals; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000737:chemistry; D052179:Antigens, Plant / Q000737:chemistry* / Q000235:genetics; D017357:Cyclotrons; D041961:Deuterium Exchange Measurement / Q000379:methods*; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000737:chemistry* / Q000235:genetics; D005583:Fourier Analysis; D006041:Goats; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D021122:Protein Subunits; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D000485:Allergens; C478880:Ana o 2 allergen, cashew; D000936:Antigen-Antibody Complex; D052179:Antigens, Plant; D000939:Epitopes; D010446:Peptide Fragments; D021122:Protein Subunits