| Autor: |
da Silva ES; Department of Biointeraction, Federal University of Bahia, Salvador, Brazil., Pacheco LGC; Department of Biointeraction, Federal University of Bahia, Salvador, Brazil., Fernandes AMS; Department of Biointeraction, Federal University of Bahia, Salvador, Brazil., Asam C; Department of Molecular and Cellular Anatomy, University of Regensburg, Regensburg, Germany., Silveira EF; Department of Biointeraction, Federal University of Bahia, Salvador, Brazil., da Silva Pinheiro C; Department of Biointeraction, Federal University of Bahia, Salvador, Brazil., Alcantara-Neves NM; Department of Biointeraction, Federal University of Bahia, Salvador, Brazil. neuzalcantara@gmail.com.; Laboratório de Alergia e Acarologia, Instituto de Ciências da Saúde, Universidade Federal da Bahia, Avenida Reitor Miguel Calmon, sem nº, Canela, CEP - 40110-100, Salvador, Bahia, Brazil. neuzalcantara@gmail.com. |
| Abstrakt: |
Successful research in the wide-ranging field of allergy is usually achieved by definition not only of physicochemical and immunological properties of natural, but also recombinant allergens. Blomia tropicalis mite is a well-known source for various groups of hypersensitivity-causing proteins. The goal of the present work was to produce, purify and characterise by in silico, biochemical and immunological methods the recombinant group-12 allergen of B. tropicalis. The recombinant Blo t 12 aggregation capacity as well as the affinity to antibodies from BALB/c immunised mice and B. tropicalis-sensitised human donors were investigated through in silico analyses, dynamic light scattering, SDS-PAGE, ELISA and Western blot. The presence of Blo t 12 within B. tropicalis extracts was also determined by ELISA and Western blot. High concentrations of dimeric rBlo t 12 were detected through SDS-PAGE next to other aggregates and the results were confirmed by data from DLS and Western blot. The YITVM peptide was predicted to be the most aggregation-prone region. The IgE-reactivity of rBlo t 12 was not completely abolished by aggregate formation but it was significantly decreased compared to rBlo t 5, or B. tropicalis extracts. Natural Blo t 12 may naturally dimerises, but it was detected in non-delipidified B. tropicalis extracts in low amounts. Given that this allergen may be a specific marker for B. tropicalis allergy, the recombinant Blo t 12 herein obtained is characterised as a mid-tier allergen in Brazilian atopic patients and may be useful for the improvement in precision allergy molecular diagnostic applications. |
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