Peanut digestome: Identification of digestion resistant IgE binding peptides

Food Chem Toxicol. 2017 Sep;107(Pt A):88-98. doi: 10.1016/j.fct.2017.06.029. Epub 2017 Jun 17.


Stability to proteolytic degradation in the digestive tract is considered a general feature shared by most food allergens. Current digestibility models exclusively utilize purified allergen proteins, neglecting the relevant effects of matrix that occur for foodstuff systems. In the present study, we investigated digestion stability of the major peanut allergens directly in the natural matrix using an in vitro static model that simulates the gastrointestinal digestion including the oral, gastric, duodenal and intestinal (brush border membrane enzymes) phases. Immunogenicity was evaluated by Western Blot using N=8 pooled sera of peanut allergic pediatric subjects. Immunoreactive, large-sized and fragments of Ara h 2, Ara h 6 and Ara h 3 survived hydrolysis as assessed by SDS-PAGE. Smaller resistant peptides mainly arising from Ara h 3 and also Ara h 1 were detected and further identified by LC-high resolution-MS/MS. RP-HPLC purification followed by dot-blot analysis and MS/MS-based identification demonstrated that stable IgE-binding peptides derived from Ara h 3. These results provide a more realistic picture of the potentially allergenic determinants of peanuts that survived the human digestion, taking into account the role of the food matrix, which may significantly affect gastrointestinal breakdown of peanut allergens.

Keywords: Allergy; Brush border membrane enzymes; IgE binding peptide; In vitro human digestion; Mass spectrometry-based proteomics; Peanut.

MeSH terms

  • Amino Acid Sequence
  • Antigens, Plant / chemistry
  • Antigens, Plant / genetics
  • Antigens, Plant / immunology
  • Antigens, Plant / metabolism
  • Arachis / chemistry*
  • Arachis / genetics
  • Arachis / immunology
  • Arachis / metabolism
  • Digestion
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Immunoglobulin E / immunology*
  • Molecular Sequence Data
  • Peanut Hypersensitivity / immunology*
  • Peanut Hypersensitivity / metabolism
  • Peptide Mapping
  • Peptides / chemistry
  • Peptides / genetics
  • Peptides / immunology*
  • Tandem Mass Spectrometry


  • Antigens, Plant
  • Peptides
  • Immunoglobulin E