Molecular and immunological characterisation of peanut ingredients for double blind placebo controlled food challenge (DBPCFC)

Abstract

Background Double blind placebo controlled food challenge (DBPCFC) is the gold standard for food allergy diagnosis and outcome measure. However, one difficulty in performing DBPCFC is the lack of well-characterised and validated sources of allergens for incurrence into the challenge food vehicle. Here, we discuss the requirements for this characterisation using the peanut flour used during the EuroPrevall project and in ongoing studies. Methods 2-D PAGE and mass spectrometry (Thermo Fisher Orbitrap Elite) was used to provide a molecular characterisation of allergens in peanut flour. Relative quantitation was used to determine batch-to-batch variation in allergen content. Immunoreactivity of the peanut flour material was assessed using rabbit polyclonal antisera raised to purified peanut allergens. Results