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Epitope mapping of 7S cashew antigen in complex with antibody by solution‐phase H/D exchange monitored by FT‐ICR mass spectrometry
Author(s) -
Guan Xiaoyan,
Noble Kyle A.,
Tao Yeqing,
Roux Kenneth H.,
Sathe Shridhar K.,
Young Nicolas L.,
Marshall Alan G.
Publication year - 2015
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.3589
Subject(s) - chemistry , epitope , monoclonal antibody , fourier transform ion cyclotron resonance , mass spectrometry , recombinant dna , epitope mapping , antigen , chromatography , microbiology and biotechnology , antibody , biochemistry , genetics , gene , immunology , biology
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. Copyright © 2015 John Wiley & Sons, Ltd.