Development of Protein and Peptide-based Micro-arrays for Peanut and Tree Nut Allergy Diagnosis

We propose to identify IgE- and IgG4-binding peptides andcombinations of peptides and intact proteins that are diagnostic of an allergic reaction to peanuts and tree nuts by peptide and protienmicroarray analysis withnut allergicserafollwed by rigorous statistical analysis of the resultant data.The specific goals and objectivesfor the project are:Goal 1) To identify IgE and IgG4 binding peptides with sera from clinically characterized nut allergic individuals.Objective: Toprobe microarrays containing synthetic overlapping peptides,from knownpeanut and tree nut (i.e. walnuts, cashew pistachio, etc) allergens, with clinically well-characterized serafrom allergic individuals in order to identify IgE and IgG4 binding sites. Goal 2) Identify clinically important allergens and the relative contribution of conformational IgE and IgG4 epitopes. Objective: Toprobe microarrays containing intact allergic proteins (or components),frompeanut and tree nut (i.e. walnuts, cashew pistachio, etc), with clinically well-characterized serafrom allergic individuals in order to identify IgE and IgG4 binding proteins.Goal 3) To have professional bioinformatics and statistical analysis of the data from Goals 1 and 2. Objective:collaborate withproffessional statisticians to support us in evaluating our methods, consult on design, prepare analysis data sets, and perform in-depth statistical analysis toidentify the most promising diagnostic peptides/proteins.Goal 4) Design new peptide/protein combination microarrays and test with allergic patient sera to confirm diagnostic value. Objective:to probe microarrays, containing only the best diagnostic peptides and intact nut allergic proteins, with clinically well-diagnosed patient serain order to test and validate the diagnositc value.