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.
Free and membrane bound peptides were synthesized based on an existing computational method, and structural database for allergic proteins, while Agricultural Research Service (ARS) has compared and assessed the allergic antibody interaction with these peptides using human serum from allergic individuals. We have cloned and expressed peanut allergens Ara h 2, a truncated Ara h 2 and Ara h 1 leader, walnut allergens Jug r 2, and Jug r 2 leader sequences in escherichia (E.) coli towards developing immunological assays and structures. Recombinant allergens will be utilized to identify clinically relevant cross-reactive epitopes (the regions of proteins associated with antibody binding) in walnut and peanut. Also, we have developed molecular models for Ara h 1, Ara h 2, and Jug r 2 leader sequences to complement our wet lab structural experiments.