Evaluation of 3 Common Methods for Effective Housefly Allergen Extraction
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Abstract
Background: Allergen extracts have been applied to treat allergic diseases. Accordingly, a housefly (Musca domestica) extract is commonly used to treat patients severely allergic to housefly.
Objective: To evaluate 3 common methods, including grinding, sonication, and homogenization, for effective preparation of housefly allergen extracts.
Methods: Housefly allergens were extracted from Musca domestica using 3 different methods, including grinding, sonication, and homogenization. Protein concentrations and profiles in the extracts were determined by Bradford assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively.
Results: The protein concentrations of the extracts prepared by grinding (mean [SD], 911.3 [159.7] µg/µL) and sonication (mean [SD], 905.7 [188.6] µg/µL) as measured by Bradford assay were significantly higher than those prepared by homogenization (mean [SD], 674.5 [60.0] µg/µL). Moreover, SDS-PAGE showed more protein bands in the extracts prepared using grinding and sonication compared to those prepared using homogenization.
Conclusions: In comparison to homogenization, both grinding and sonication methods are superior ways to prepare housefly allergen extracts as evidenced by the higher quantities and composition of proteins.
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References
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