Purification and characterization ofLep dI, a major allergen from the miteLepidoglyphus destructor

摘要

SummaryA major allergen of the storage miteLepidoglyphus destructor (Lep dI) has been purified by affinity chromatography using an anti‐Lep dI monoclonal antibody. The purity of the protein obtained by this procedure was assessed by reverse‐phase HPLC.Lep dI displayed a molecular weight of 14 kD on SDS‐PAGE under non‐reducing conditions, and 16 kD in the presence of a reducing agent. Analytical IEF revealed a little charge microheterogeneity, showing three bands with pIs 7.6–7.8. PurifiedLep dI retained IgE‐binding ability, as proved by immunoblotting experiments after SDS‐PAGE and RAST with individual sera fromL. destructor‐sensitive patients. Results from the latter technique demonstrated that 87 ofL. destructor‐allergic patients had specific IgE toLep dI, and a good correlation between IgE reactivity withL. destructorextract andLep dI was found. In addition, RAST inhibition experiments showed that IgE‐binding sites onLep dI are majorL. destructor‐allergenic determinants, sinceLep dI could inhibit up to 75 the binding of specific IgE toL. destructorextract; on the other hand.Lep dI did not cross‐react withD.