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Isolation and characterization of the allergen Dpt 12 from Dermatophagoides pteronyssinus by chromatofocusing
Abstract:
The allergen Dpt 12, from the house dust mite Dermatophagoides pteronyssinus, was isolated from spent growth medium by chromatofocusing. The allergen was isolated in the pI range 5.86-6.95, indicating heterogeneity with regard to pI. However, on sodium dodecylsulphate-polyacrylamide gel electrophoresis the allergen appeared homogeneous with an apparent molecular weight of 27,000 daltons. Radioallergosorbent scores utilizing paper discs coupled with Dpt 12 correlated significantly (p less than 0.0005) with those obtained using discs coupled with whole mite extract, although the corresponding titres were higher with Dpt 12 (p less than 0.0005).
Insights
Researchers isolated the Dpt 12 allergen from house dust mites. This purified allergen showed significant correlation with whole mite extract in allergy tests, suggesting its potential for diagnostics.
Area of Science:
- Immunology
- Allergology
- Biochemistry
Background:
- House dust mites (Dermatophagoides pteronyssinus) are a common source of allergens.
- Understanding specific mite allergens is crucial for diagnosing and treating allergic diseases.
Purpose of the Study:
- To isolate and characterize the Dpt 12 allergen from Dermatophagoides pteronyssinus.
- To evaluate the allergenic potential of purified Dpt 12 in comparison to whole mite extract.
Main Methods:
- Isolation of Dpt 12 using chromatofocusing.
- Characterization of allergen purity and molecular weight via sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE).
- Assessment of allergenic activity using radioallergosorbent tests (RAST).
Main Results:
- Dpt 12 was isolated with a pI range of 5.86-6.95, indicating heterogeneity.
- SDS-PAGE revealed a homogeneous protein with an apparent molecular weight of 27,000 daltons.
- RAST scores using Dpt 12 showed significant correlation (p < 0.0005) with whole mite extract, with higher titres observed for Dpt 12.
Conclusions:
- The Dpt 12 allergen from Dermatophagoides pteronyssinus can be purified and characterized.
- Purified Dpt 12 demonstrates significant allergenic activity, comparable to whole mite extracts.
- Dpt 12 is a valuable component for specific immunotherapy and diagnostic assays for house dust mite allergy.