Recombinant Allergen-Based Quantum Dot Lateral Flow Immunoassays for Component-Specific IgE Detection in House Dust Mite Allergy

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Abstract

House dust mite (Dermatophagoides pteronyssinus) allergy is commonly evaluated by serum allergen-specific IgE (sIgE), but assays based on natural extracts are limited by compositional complexity and batch variability. We prepared recombinant Der p 1, Der p 2, Der p 10, and Der p 23 and evaluated their suitability as standardized component-resolved diagnostic antigens. The allergens were analyzed by bioinformatics, expressed in Escherichia coli, purified, and characterized by circular dichroism spectroscopy and sedimentation-velocity analytical ultracentrifugation. Recombinant proteins were then incorporated as capture antigens in a quantum dot-based lateral flow immunoassay (QD-LFIA) for quantitative sIgE measurement, and performance was compared with ImmunoCAP using residual clinical serum samples. The four allergens showed distinct physicochemical and structural features and different distributions of predicted B-cell linear epitopes. All four proteins were successfully expressed and purified, and structural analyses supported predominantly monomeric behavior in solution. The assay showed linear responses from 0.35 to 100 IU/mL (R² ≥ 0.9935). In component-specific clinical comparisons, QD-LFIA and ImmunoCAP results were strongly correlated (R² = 0.9718–0.9894), and more than 95% of paired observations fell within the calculated Bland–Altman limits of agreement. These findings establish a workflow linking recombinant allergen preparation and structural quality assessment with rapid component-specific sIgE measurement and support further validation of recombinant Der p allergens for standardized house dust mite allergy testing.

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