Date

Thursday 12 Mar 2026

Time

16:00 - 16:30

Location

BW006

Supervisor

Sander van Kasteren

Jury

Rolf Boot

External supervisor

Menno van Zelm

Component-resolved serology can quantify food allergen sensitization, but it does not reliably predict clinically relevant food allergy and oral food challenges (OFCs) remain the gold standard. Here, we validated a panel of in-house produced recombinant food allergen components for IgE serology and as input for a functional flow cytometry-based assay (CytoBas).

Thirteen allergen components from peanut, cashew, hazelnut, cow’s milk, hen’s egg, and wheat were recombinantly produced in-house. Component-specific IgE was measured by in-house ELISA and benchmarked against state-of-the-art multiplex IgE serology (ALEX²) in 93 pediatric patients with food allergy. ELISA–ALEX² agreement, ROC analysis for evaluation of diagnostic performance and multi-component approaches (OR rules, logistic regression) were performed.

For peanut, hazelnut, and milk, rAra h 2 (AUC = 0.93, sensitivity 89.8%, specificity 90.7%), rCor a 14 (AUC = 0.90, sensitivity 84.1%, specificity 91.1%), and rBos d 9 (AUC = 0.98, sensitivity 100%, specificity 94.6%) were the strongest single-component discriminators between clinically allergic and non-allergic children respectively. For egg, rGal d 1 was the best component (AUC = 0.84, sensitivity 75.0%, specificity 93.5%). For cashew, rAna o 1 and rAna o 2 were insufficient (AUC = 0.58–0.65, sensitivity 22.8–31.6%, specificity 93.3-100%). Wheat showed high performance for Tri a 19 (AUC = 0.94; sensitivity 100%; specificity 88.9%), but remained inconclusive (allergic patients n = 2). Most components demonstrated comparable ELISA and ALEX² AUCs (e.g., Ara h 2, Cor a 14, Bos d 9), whereas Ara h 1, Bos d 5, Cor a 11, Ana o 2, Gal d 2, and Gal d 3 showed significant platform-dependent differences (p<0.05). OR-based combinations improved sensitivity but reduced specificity when lower-specificity components were included, logistic regression did not improve performance.

Overall, serum component-specific IgE reflected sensitization but only partially aligned with clinical allergy. For several components, ELISA and ALEX² yielded significantly different AUCs, demonstrating that diagnostic performance depends on both the selected targets and the serological platform used. Because serum IgE and basophil FcεRI-bound IgE represent distinct IgE pools, Functional validation of these components in a flow cytometric basophil-bound IgE assay is required to determine effector-level relevance and finalize panel refinement.