1a.Objectives (from AD-416)
Use empirical tools to improve computational prediction models for detecting clinically relevant linear IgE binding sites.
1b.Approach (from AD-416)
a. Synthesize known and computationally predicted cross-reactive IgE binding epitopes of peanuts and tree nuts on a membrane.
b. Use clinically well-characterized patient sera with known peanut and/or tree-nut (walnut and almond or cashew) allergy to assess the clinical relevance of IgE binding to a particular synthetic peptide.
c. Continuously optimize computational model based on empirical data obtained from goal b.
We have cloned and expressed peanut allergenic proteins Ara h 2, a truncated Ara h 2 and the Ara h 1 leader (leading amino acids of the allergen peptide), walnut allergens Jug r 2 and the Jug r 2 leader sequences in E. coli to help identify their structures. We have also developed molecular models for Ara h 1, Ara h 2 and the Jug r 2 leader sequences to complement our structural experiments. This project was monitored by standard email, telephone conversations and by occasional meetings.