Print Email Facebook Twitter Purification and immunoglobulin E-binding properties of peanut allergen Ara h 6: Evidence for cross-reactivity with Ara h 2 Title Purification and immunoglobulin E-binding properties of peanut allergen Ara h 6: Evidence for cross-reactivity with Ara h 2 Author Koppelman, S.J. de Jong, G.A.H. de Laaper-Ertmann, M. Peeters, K.A.B.M. Knulst, A.C. Hefle, S.L. Knol, E.F. TNO Kwaliteit van Leven Publication year 2005 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 14981 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. © 2005 Blackwell Publishing Ltd. Chemicals / CAS: immunoglobulin E, 37341-29-0; Albumins; Allergens; Ara h 2 allergen, Arachis hypogaea; Ara h 6 allergen, Arachis hypogaea; Glycoproteins; Immunoglobulin E, 37341-29-0; Plant Proteins; Recombinant Proteins Subject Nutrition SafetyInnovation PolicyAllergenBasophilIgEPeanutProtein purificationSkin prick testallergenAra h 2 allergenAra h 6 allergenepitopeimmunoglobulin Emonomerpeanut agglutininunclassified drugadultamino acid sequenceamino terminal sequenceantigen antibody reactionantigen bindingarticlebasophil degranulationcarboxy terminal sequenceclinical articleclinical trialcontrolled clinical trialcontrolled studycross reactiondouble blind procedurehumanhuman cellimmunoblottingmatrix assisted laser desorption ionization time of flight mass spectrometrymolecular weightpeanut allergyprick testpriority journalprotein analysisprotein purificationsequence homologyAdultAlbuminsAllergensAmino Acid SequenceAntibody SpecificityBasophilsCross ReactionsGlycoproteinsHumansHypersensitivityImmunoglobulin EMolecular WeightPlant ProteinsRecombinant Proteins To reference this document use: http://resolver.tudelft.nl/uuid:e9ab9071-baa6-414e-b319-e30f97b2c484 DOI https://doi.org/10.1111/j.1365-2222.2005.02204.x TNO identifier 238425 ISSN 0954-7894 Source Clinical and Experimental Allergy, 35 (4), 490-497 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.