Human IgE against the major allergen Bet v 1--defining an epitope with limited cross-reactivity between different PR-10 family proteins.
Bottom Line: This provides a potential explanation, at the molecular level, for the differences in recognition of isoforms of Bet v 1 and other allergens in the PR-10 protein family displayed by IgE targeting this epitope.Finally, we present the first high-resolution structure of a human allergen-specific IgE fragment in the single-chain fragment variable (scFv) format.Such studies may aid us in development of better diagnostic tools and guide us in the development of new therapeutic compounds.
Affiliation: Department of Immunotechnology, Lund University, Lund, Sweden.Show MeSH
Related in: MedlinePlus
Mentions: To enable characterization of molecular interactions between antibody fragments with origin in the human IgE repertoire and the major birch pollen Bet v 1, we performed phage display selections on the three Bet v 1 isoforms Bet v 1.0101, Bet v 1.0102 and Bet v 1.0112 from an antibody fragment library created from atopic donors. The library was created from transcripts encoding the VH domain of IgE isolated from nasal tissue of two allergic subjects known to have an IgE repertoire overrepresented by antibodies derived from the IGHV5 gene subgroup 4. These selections generated four novel-specific binders (B10, B13, B14 and M0418). Three binders, B10, B13 and B14, were isolated by selection on the major isoform Bet v 1.0101, while selection on isoform Bet v 1.0102 generated one additional binder (M0418) together with re-isolation of binder B14. Selections performed on isoform Bet v 1.0112 failed to generate any binders. All isolated scFv were highly Bet v 1-specific, showing no cross-reactivity to a panel of unrelated allergens (Fig.1).
Affiliation: Department of Immunotechnology, Lund University, Lund, Sweden.