da Silva, Eduardo Santos and Carvalho Pacheco, Luis Gustavo and Santana Fernandes, Antonio Marcio and Asam, Claudia and Silveira, Elisania Fontes and Pinheiro, Carina da Silva and Alcantara-Neves, Neuza Maria (2021) Purification and characterisation of the dimeric group 12 allergen from Blomia tropicalis heterologously expressed by Escherichia coli Top10F '. MOLECULAR BIOLOGY REPORTS, 48 (4). pp. 3405-3416. ISSN 0301-4851, 1573-4978
Full text not available from this repository. (Request a copy)Abstract
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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | DUST MITE ALLERGEN; CRYSTAL-STRUCTURE; AGGREGATION; CELLS; Aggregation; rBlo t 12; nBlo t 12; Dimer; House dust mite; Mite extract |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Anatomie > Lehrstuhl für Molekulare und zelluläre Anatomie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 14 Sep 2022 09:19 |
| Last Modified: | 14 Sep 2022 09:19 |
| URI: | https://pred.uni-regensburg.de/id/eprint/47436 |
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