Actual questions of standardization of water-salt extracts of house dust mites mixed allergens intended for medicinal forms of sublingual immunotherapy

Abstract

Background: Standardization of medical forms of allergens is the process of establishing and applying a unified system of quality indicators and control methods. Quality control of therapeutic forms of allergens is an essential condition for guaranteeing the clinical efficacy and safety of allergen-specific immunotherapy (ASIT). The technology for obtaining the granular form of the mixed allergen from house dust mites Dermatophagoides pteronyssinus (Der p) and Dermatophagoides farinae (Der f) for oral use was developed in the laboratory of allergens Mechnikov Research Institute of Vaccines and Sera. Aims: to determine the authenticity of the anphenolic water-salt extract of the mixed allergen from mites Der p and Der f, intended for the therapeutic form of the drug. Materials and methods: Standardization of the obtained allergen extract was carried out by protein nitrogen unit by the Nessler method. The protein concentration was determined by the Bradford method. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to study the presence of protein composition in mixed allergen. The pH value was determined potentiometrically. The comparison of the specific activity of the standard allergen and the experimental mixed allergen was carried out by the method of reverse immunoassay (REAST). Results: SDS-PAGE showed the preservation of the protein profile of the water-salt extract of the mixed allergen Der p and Der f, corresponding to the protein profile of allergens of house dust mites. There were no statistically significant differences in the optical density of the studied allergen (four drug series) and the standard allergen with reference serums containing specific IgE to Der p or Der f when reacting with a Class 3 reference serum in REAST(р0,05). Conclusions: The above in accordance with the State Pharmacopoeia of the Russian Federation XIV edition is a confirmation of the authenticity of the drug. Not phenolic extraction did not affect the main allergenic components of the mixed allergen Der p and Der f and allowed it to maintain a high degree of specificity.