With the evolution of peptide synthesis techniques and microarray technology, it is now possible to map and characterize allergenic
epitopes on a microarray platform. The
peptide microarray-based immunoassay allows simultaneous analysis of thousands of target
peptides using small volumes of diluted serum samples, and has a promising future to become a superior testing tool for aspects of
food allergy diagnosis and prognosis, as well as for designing recombinant
allergens for safe
immunotherapy. Characterization of allergenic
epitopes provides fundamental knowledge for understanding mechanisms of
food allergy. This chapter describes in detail the development of a sensitive and reliable
peptide microarray-based immunoassay. The information includes a comparison of different slide substrates, effect of
buffers on spot morphology, performance of printing pins, immunolabeling detection systems with different levels of sensitivity, and suggested approaches to data analysis.