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

BACKGROUND: Standardization of medical forms of allergens is the process of establishing and applying a unified system of quality indicators and control methods. The quality control of therapeutic forms of allergens is an essential condition for guaranteeing the clinical efficacy and safety of allergen-specific immunotherapy. The technology for obtaining the granular form of the mixed allergen from house dust mites Dermatophagoides pteronyssinus and Dermatophagoides farinae for oral use was developed in the laboratory of allergens Mechnikov Research Institute of Vaccines and Sera.

AIM: To determine the authenticity of the anphenolic water-salt extract of the mixed allergen from mites D. pteronyssinus and D. farinae, intended for the therapeutic form of the drug.

MATERIALS AND METHODS: Standardization of the obtained allergen extract was performed by protein nitrogen unit using the Nessler method. The protein concentration was determined using the Bradford method. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis was used to study the presence of protein composition in mixed allergen. The pH value was determined potentiometrically. The specific activity of the standard allergen and the experimental mixed allergen were compared using the reverse immunoassay method.

RESULTS: Sodium dodecyl sulfate–polyacrylamide gel electrophoresis showed the preservation of the protein profile of the water-salt extract of the mixed allergen D. pteronyssinus and D. farinae, corresponding to the protein profile of allergens of house dust mites. No statistically significant differences were noted in the optical density of the studied allergen (four drug series) and the standard allergen with reference serums containing specific IgE to D. pteronyssinus or D. farinae when reacting with a class 3 reference serum in the reverse immunoassay method (р >0.05).

CONCLUSIONS: The result of the study 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 D. pteronyssinus and D. farinae and allowed it to maintain a high degree of specificity.