Detection Of Peanut Allergens In Breast Milk and Saliva
AAAI Annual Meeting San Diego CA March 2014
Infantshave been reported to react to peanut upon their first known exposure and peanut proteinshave been detected in breast milk. Identification of the allergens or fragments thereof in breast milk may allow us todetermine if they are sensitizing ortolerizing.Methods:
Variousimmunoassayswere optimized andutilized to analyze forthe presence of peanut proteins in breast milk. Breast milk samples from healthy lactating motherswere collected following a 48 hour peanut fast and spiked with known amounts of peanuts and subjected toimmunoprecipitation, SDS-PAGE, western blot and ELISA with anti-peanut, anti-Ara h 1, 2 and 3 antibodies. Mass spectrometrywas performed toidentify peanut peptides in breast milk.Results:
We found that we were able to detect peanut allergen, Ara h 1, Ara h 2 and Ara h3, in peanut-spiked breast milk and saliva at nM levels. We were able to identify specific peanut peptides of Ara h 1, Ara h 2 and Ara h 3/4 using mass spectrometry.
Conclusions: The fact that allergic proteins or peptides survive in digestive enzymes, and are likely to be secreted in biological fluids indicates they are most likely the sensitizing or tolerizing agents within an allergic food. Developing methods to detect these allergens in breast milk is a preliminary step in identification of allergens or fragments thereof that are secreted and may contribute to the original development of allergic disease.