D3.104 - Molecular Co-Sensitization Mapping Using a 192-Line Multiplex IgE Platform

High-density multiplex IgE assays enable simultaneous measurement of extract and molecular component allergens, allowing system-level characterization of sensitization patterns.

The PROTIA Allergy-Q 192D is a 192-line immunoblot platform capable of analyzing 176 allergen-specific IgE analytes, including molecular components, extracts, drug/chemical allergens, and total IgE within a single assay. While analytical agreement is essential, the ability of a multiplex system to preserve biologically structured co-sensitization architecture across numerous analytes represents a critical indicator of technical robustness.

This study aimed to evaluate whether the 192-line platform reproducibly identifies coherent molecular co-sensitization modules independent of clinical data.

A total of 263 serum samples were analyzed using the PROTIA Allergy-Q 192D multiplex line immunoassay. IgE positivity was defined at a threshold equivalent to ≥0.35 kU/L.

Allergen-specific IgE responses were evaluated at both individual analyte and protein family levels. Polysensitization burden was calculated as the number of positive analytes per subject. Conditional co-sensitization probabilities were assessed for selected index components, including representative PR-10 and profilin allergens.

Pairwise associations between analytes were quantified using phi coefficients derived from binary positivity matrices. Network construction was performed by applying correlation thresholds to identify significant edges, and modularity was calculated using the Louvain community detection algorithm. Subject-level sensitization phenotypes were explored through hierarchical clustering based on Jaccard distance.

Clinical symptom data were not included in the analysis.

Sensitization to at least one analyte was observed in 100% of samples, with a median of 51 positive analytes per subject (IQR 18–94), demonstrating broad polysensitization detectable by the high-density format.

Conditional analysis revealed distinct molecular signatures. PR-10 index positivity was associated with strong intra-family co-sensitization and limited cross-family overlap, whereas profilin positivity demonstrated broader cross-pollen associations.

Phi correlation analysis identified pronounced intra-family connectivity among homologous components. Network modeling demonstrated discrete allergen modules with modularity Q ≥ 0.16, indicating structured molecular clustering rather than random co-positivity.

Hierarchical clustering further delineated reproducible sensitization phenotypes, including PR-10-dominant, mite/pet-dominant, and multi-component polysensitized patterns.

The 192-line multiplex IgE platform enables high-resolution mapping of molecular co-sensitization architecture across 176 analytes, including total IgE. Structured modular clustering confirms that high-density multiplexing preserves biologically coherent sensitization patterns, supporting its utility for advanced molecular epidemiology and large-scale sensitization profiling.