FIGURE 1.

Generation of nanobody‐based human IgE against nApi m 1 and rApi m 2 (A, B) Schematic representation of the nanobody generation. Upon immunization of llamas with nApi m 1 and rApi m 2, immune repertoire libraries were generated and selected by phage display. Nanobody‐based human IgEs (nb‐IgEs) were then generated by fusion of obtained allergen‐specific nanobodies to the Fc domains of human IgE. (C) SDS‐PAGE and immunoblotting analysis of purified nb‐IgEs under non‐reducing (NR) and reducing (R) conditions. The recombinant nb‐IgE were detected using an anti‐IgE antibody conjugated to alkaline phosphatase. (D) The immunoreactivity of purified nb‐IgEs was further assessed in ELISA. Binding to immobilized allergen or FcεRIα was detected using an anti‐IgE antibody conjugated to alkaline phosphatase. Data are mean ± s.d. of triplicates