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Development of high-affinity, single-domain protein binders for neutralizing household allergens

Zhao, E. M.; Zhang, D. K.; Yue, Q.; He, Q.; Li, Y.; Qu, Z.; Zhao, G.; Andreissen, N.

2025-08-03 bioengineering
10.1101/2025.08.03.668213 bioRxiv
Show abstract

Feline, canine, and human dust mite allergens drive significant IgE-mediated allergies in the household environment. Here, we describe the discovery and characterization of novel protein binders, termed AVA, which efficiently bind and disrupt major allergens from cats (Fel d 1), dogs (Can f 1, Can f 2), and dust mites (Der p 1, Der p 2). Leveraging camelid single-domain variable-heavy chain (VHH) antibodies, we identified and optimized VHH sequences with robust affinity towards the individual allergens. AVA disrupted critical allergen substructures (Fel d 1) and catalytic functions (Der p 1) in molecular dynamics simulations and in in vitro assays, respectively. Paired with robust thermostability and non-toxic in vitro viability profiles, AVA represents a promising approach for downstream household allergen mitigation.

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