Designing a multimer allergen for diagnosis and immunotherapy of dog allergic patients.

Abstract

BackgroundDog dander extract used for diagnosis and allergen-specific immunotherapy is often of variable and of poor quality.ObjectiveTo assemble four well-established dog allergen components into one recombinant folded protein for improved diagnosis and vaccination of allergy to dog.MethodsA linked molecule, comprising the four dog lipocalin allergens Can f 1, Can f 2, Can f 4 and Can f 6 was constructed. The tetrameric protein was structurally characterized by small angle X-ray scattering, and compared with each single recombinant lipocalin allergen or an equimolar mix of the four allergens by analytical size exclusion chromatography, circular dichroism, allergen-specific IgE in serum by ELISA and allergen-dependent capacity to activate basophils. The immunogenicity of the fusion protein was evaluated in immunized mice by assessing splenocyte proliferation and antibody production.ResultsThe linked tetrameric construct was produced as a soluble fusion protein, with the specific folds of the four individual allergens conserved. This multi-allergen molecule was significantly more efficient (pConclusionWe provide the first evidence for a linked recombinant molecule covering the major dog allergens for potential use in diagnostics and allergy vaccination of dog allergic patients.