Purification and characterization of Lep d I, a major allergen from the mite Lepidoglyphus destructor.
A major allergen of the storage mite Lepidoglyphus destructor (Lep d I) has been purified by affinity chromatography using an anti-Lep d I monoclonal antibody. The purity of the protein obtained by this procedure was assessed by reverse-phase HPLC. Lep d I 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. Purified Lep d I retained IgE-binding ability, as proved by immunoblotting experiments after SDS-PAGE and RAST with individual sera from L. destructor-sensitive patients. Results from the latter technique demonstrated that 87% of L. destructor-allergic patients had specific IgE to Lep d I, and a good correlation between IgE reactivity with L. destructor extract and Lep d I was found. In addition, RAST inhibition experiments showed that IgE-binding sites on Lep d I are major L. destructor-allergenic determinants, since Lep d I could inhibit up to 75% the binding of specific IgE to L. destructor extract; on the other hand, Lep d I did not cross-react with D. pteronyssinus allergens.
Ventas
P
P
Departamento de Investigación, Alergia e Inmunología Abelló S.A., Madrid, Spain.
Carreira
J
J
Polo
F
F
D000485:Allergens / Q000737:chemistry / Q000302:isolation & purification*; D000818:Animals; D002846:Chromatography, Affinity; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007120:Immunochemistry; D007073:Immunoglobulin E / Q000097:blood; D007526:Isoelectric Point; D008925:Mites / Q000276:immunology*; D011852:Radioallergosorbent Test
D000485:Allergens; D007073:Immunoglobulin E