%0 Generic %A F., Schocker %A A., Scharf %A S., Kull %A U., Jappe %D 2017 %T Supplementary Material for: Detection of the Peanut Allergens Ara h 2 and Ara h 6 in Human Breast Milk: Development of 2 Sensitive and Specific Sandwich ELISA Assays %U https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Detection_of_the_Peanut_Allergens_Ara_h_2_and_Ara_h_6_in_Human_Breast_Milk_Development_of_2_Sensitive_and_Specific_Sandwich_ELISA_Assays/5441530 %R 10.6084/m9.figshare.5441530.v1 %2 https://karger.figshare.com/ndownloader/files/9409003 %2 https://karger.figshare.com/ndownloader/files/9409006 %2 https://karger.figshare.com/ndownloader/files/9409009 %K Peanut allergy %K Ara h 2 %K Ara h 6 %K Sandwich ELISA %K Breast milk %X

Background: Little is known about breast milk as a vehicle for tolerance development or sensitization to peanuts very early in life. Thus, well-characterized and highly sensitive detection systems for the reliable determination of peanut allergens in breast milk are mandatory. Methods: For the quantification of the marker allergens Ara h 2 and Ara h 6 in the low nanogram per milliliter range in breast milk samples of a German cohort, sensitive and highly specific sandwich ELISAs were optimized and validated. Results: The Ara h 2 ELISA revealed a limit of detection (LOD) of 1.3 ng Ara h 2/mL and a quantification range of 2.3-250 ng/mL, the Ara h 6 ELISA showed an LOD of 0.7 ng/mL and a working range of 1.1-14.4 ng/mL. The assays showed no relevant cross-reactivity against other potentially cross-reactive legume, seed, and tree nut extracts (<0.01%, except for Ara h 1 in the Ara h 2 ELISA <0.1%). Ara h 2 was detectable in breast milk samples from 14/40 (35%) of the participants in concentrations from 2.3 to 184 ng/mL, Ara h 6 appeared in 9/40 (22.5%) of the lactating mothers between 1.1 and 9.7 ng/mL, and 1 highly positive sample with 79 ng/mL. Both allergens appeared at the same time points, but Ara h 6 in lower concentrations than Ara h 2. Conclusions: Sensitive and specific diagnostic tools for the determination of Ara h 2 and Ara h 6 in human breast milk were established. The kinetics of secreted Ara h 2 and Ara h 6 seem to be similar but with a difference in concentration. Follow-up investigations on their tolerogenic or sensitizing properties in breast milk become now accessible.

%I Karger Publishers