1. Production of crud allergen extract from Aspergillus fumigatus and recombinant Asp f 1 in Escherichia coli, and compare their immunological characterization.
- Author
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NIKPOUR, SABOURA, SAFARPOUR, HOSSEIN, RAFIEE, MITRA, and FEREIDOUNI, MOHAMMAD
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ALLERGENIC extracts , *ASPERGILLUS fumigatus , *ESCHERICHIA coli , *ALLERGIES , *RECOMBINANT proteins - Abstract
The prevalence of allergic diseases and asthma is increasing worldwide. Fungi have been identified as one of the most important inhalation allergens and can trigger allergic symptoms, particularly asthma, and rhinosinusitis. Among allergenic fungi, Aspergillus fumigatus is more important in causing allergic diseases, and its extract is usually used in skin prick tests. This study aimed to produce an allergenic extract of Aspergillus fumigatus and recombinant Asp f 1 in Escherichia coli at Birjand University of Medical Sciences. Material and methods The allergenic extract of Aspergillus fumigatus was prepared using the standard strain. rAsp f 1 allergen was expressed in Escherichia coli BL21 cells and purified by Ni-NTA affinity chromatography. The amount of protein in both extracts was measured by the Bradford method. SDS-PAGE was performed to evaluate the proteins of the crude extract and the purity of rAsp f 1. Allergenicity of protein was evaluated by dot blot and ELISA compared with crude extract. Results SDS-PAGE of total proteins from the crude allergen extract demonstrated several bands which included the most critical allergens of Aspergillus fumigatus. The recombinant protein rAsp f 1 was successfully expressed, followed by purification, and its purity was subsequently validated through SDS-PAGE. The reactivity of the two extracts with serum samples obtained from sensitized patients was confirmed through dot blot and ELISA. Conclusions The results of this study demonstrated that utilizing recombinant allergens as alternatives to crude allergen extracts can enhance the efficacy and specificity in detecting sensitivity to Aspergillus fungus. [ABSTRACT FROM AUTHOR]
- Published
- 2024