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Ara h 6 complements Ara h 2 as an important marker for IgE reactivity to peanut

Clinical and Translational Allergy
Springer (Biomed Central Ltd.)
Publication Date
DOI: 10.1186/2045-7022-3-s3-p168
  • Poster Presentation
  • Biology
  • Medicine


Ara h 6 complements Ara h 2 as an important marker for IgE reactivity to peanut POSTER PRESENTATION Open Access Ara h 6 complements Ara h 2 as an important marker for IgE reactivity to peanut J Hindley1*, A Koid2, RG Hamilton3, R van Ree4, SA Versteeg4, SC Dreskin5, MD Chapman2, S Wunschmann2 From Food Allergy and Anaphylaxis Meeting (FAAM 2013) Nice, France. 7-9 February 2013 Background Ara h 6 has a reported seroprevalence similar to Ara h 2, a major peanut allergen. Both allergens are of similar molecular size and share 50% identity in their amino acid sequences. Their similarity has made it difficult to obtain natural Ara h 6 free of Ara h 2 for immunoreactivity studies. The objective of this study was to obtain purified natural Ara h 6 that is essentially free of Ara h 2 and to compare its IgE reactivity and potency in histamine release assays to Ara h 2. Methods Natural Ara h 6 was purified from peanut flour extract by affinity chromatography and size exclusion chromatogra- phy. Purified Arah 6 was analyzed by silver-stained SDS- PAGE and tandem mass spectrometry (LC/MS-MS). The immunological and biological reactivity of Ara h 6 and Ara h 2 were compared by ELISA and histamine release assays. Results SDS-PAGE of the highly purified allergen revealed a single 14.5kD band and the identity of Ara h 6 was confirmed by tandem mass spectrometry (LC/MS-MS). The purified nAra h 6 contained <0.01% traces of Ara h 2 as assessed by ELISA. Natural Ara h 6 had a lower biological activity in basophil histamine release assays than natural Ara h 2. Chimeric ELISA showed that 70 and 75% of peanut aller- gic patients (n=57) had specific-IgE to natural Ara h 2 and natural Ara h 6 respectively. Conclusion Ara h 6 is a major peanut allergen, with comparable immu- noreactivity to Ara h 2. The highly purified Ara h 6, free of Ara h 2, will be useful for diagnostic IgE antibody assays, and for molecular and cellular studies to further investigate the immunological mechanisms of peanut alle

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