Journal Article

Rapid Screening for Potential Epitopes Reactive with a Polycolonal Antibody by Solution-Phase H/D Exchange Monitored by FT-ICR Mass Spectrometry

Mao, Yuan, Noble, Kyle A., Zhang, Qian, Sathe, Shridhar K., Roux, Kenneth H., Marshall, Alan G., Young, Nicolas L.

2013

Overview

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. Figure ᅟ

• Publisher: Springer-Verlag,Springer Nature B.V
• Subject: Allergens - chemistry / Allergens - genetics / Amino Acid Sequence / Anacardium - chemistry / Anacardium - genetics / Anacardium - immunology / Analytical Chemistry / Animals / Antibodies / Antibody Specificity / Antigen-Antibody Complex - chemistry / Antigens, Plant - chemistry / Antigens, Plant - genetics / Bioinformatics / Biotechnology / Chemistry / Chemistry and Materials Science / Cyclotron resonance / Cyclotrons / Deuterium Exchange Measurement - methods / Epitope Mapping - methods / Epitopes - chemistry / Epitopes - genetics / Food allergies / Fourier Analysis / Fourier transforms / Goats / Homology / Immunization / Ions / Mass spectrometry / Models, Molecular / Molecular Sequence Data / Organic Chemistry / Peptide Fragments - chemistry / Peptide Fragments - genetics / Polyclonal antibodies / Protein Subunits / Proteomics / Research Article / Scientific imaging / Spectrometry, Mass, Electrospray Ionization - methods / Spectroscopy