title: Purification and immunoglobulin E-binding properties of peanut allergen Ara h 6: evidence for cross-reactivity with Ara h 2.
abstract: BACKGROUND
  IgE-binding peanut proteins smaller than 15 kDa were previously identified as potential allergens in the majority of our peanut allergic population.
  
  
  OBJECTIVE
  To characterize the novel allergen in order to determine whether it was similar to one of the thus far identified recombinant peanut allergens (Ara h 1-7).
  
  
  METHODS
  An IgE-binding protein of <15 kDa was purified and identified via N-terminal sequencing. Its IgE-binding properties were investigated using immunoblotting, basophil degranulation, and skin prick testing. Possible cross-reacting epitopes with other peanut allergens were studied using IgE-immunoblotting inhibition.
  
  
  RESULTS
  The purified protein is a monomeric protein with a molecular weight of 14,981 Da as determined using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectroscopy. The amino acid sequence of the first 39 N-terminal residues is identical to that of Ara h 6, indicating that the allergen is Ara h 6. It is recognized by 20 out of 29 peanut-allergic patients on IgE-immunoblot, and its potent biological functionality is demonstrated by the degranulation of basophils, even at concentrations below 10 pg/mL, and by positive skin prick reactions. Ara h 6 has homology to Ara h 2, especially in the middle part and at the C-terminal part of the protein. Almost complete inhibition of IgE-Ara h 6 interaction with Ara h 2 demonstrates that at least part of the epitopes of Ara h 6 are cross-reactive with epitopes on Ara h 2.
  
  
  CONCLUSIONS
  Peanut-derived Ara h 6 is a biologically active allergen recognized by the majority of our peanut-allergic patient population and can be considered a clinically relevant peanut allergen.
authors.affiliation: TNO Nutrition and Food Research, Zeist, The Netherlands. koppelman@voeding.tno.nl
authors.forename: S J
authors.initials: SJ
authors.lastname: Koppelman
authors.forename: G A H
authors.initials: GA
authors.lastname: de Jong
authors.forename: M
authors.initials: M
authors.lastname: Laaper-Ertmann
authors.forename: K A B M
authors.initials: KA
authors.lastname: Peeters
authors.forename: A C
authors.initials: AC
authors.lastname: Knulst
authors.forename: S L
authors.initials: SL
authors.lastname: Hefle
authors.forename: E F
authors.initials: EF
authors.lastname: Knol
mesh_terms: D055516:2S Albumins, Plant; D000328:Adult; D000418:Albumins / Q000276:immunology / Q000302:isolation & purification; D000485:Allergens / Q000276:immunology* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000918:Antibody Specificity / Q000276:immunology; D052179:Antigens, Plant; D001491:Basophils / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007073:Immunoglobulin E / Q000276:immunology*; D008970:Molecular Weight; D010940:Plant Proteins / Q000276:immunology* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D055516:2S Albumins, Plant; D000418:Albumins; D000485:Allergens; D052179:Antigens, Plant; C500890:Ara h 2 allergen, Arachis hypogaea; C492737:Ara h 6 allergen, Arachis hypogaea; D006023:Glycoproteins; D010940:Plant Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E