Certification of a pharmacopoeial reference standard of lyophilized IgE-containing serum to ragweed pollen allergen

INTRODUCTION. The assessment of allergen activity by competitive enzyme-linked immunosorbent assay (ELISA) requires reference standards (RS) of specific sera containing IgE antibodies to the common ragweed pollen allergen. However, neither an international nor a national standard for such serum is available. Therefore, the development of a Russian pharmacopoeial reference standard (PhRS) for IgE-containing serum to common ragweed pollen allergen is relevant.

AIM. This study aimed to certify a candidate for the pharmacopoeial reference standard of lyophilized IgE-containing serum to common ragweed pollen allergen (PhRS IgE-Amb) and to evaluate the feasibility of its use for determining IgE concentration and allergen activity.

MATERIALS AND METHODS. The concentration of allergen-specific IgE antibodies in the candidate for the PhRS IgE-Amb was determined by reverse allergosorbent test (REAST) using a commercial reagent kit manufactured in Russia. The identity of the candidate for the PhRS IgE-Amb was determined according to General Pharmacopoeia Monograph 1.7.2.0034.15 by direct ELISA. Tests of the candidate for the PhRS IgE-Amb for the quality attributes (Description, Water, Mass homogeneity, Solubility, and Tightness) were conducted in accordance with the requirements of the State Pharmacopoeia of the Russian Federation (SP RF). Allergenic activity was assessed by competitive ELISA. Statistical analysis was performed using analysis of variance (ANOVA).

RESULTS. The candidate for the PhRS IgE-Amb was certified according to the approved certification program. The certified characteristic of the PhRS IgE-Amb, i.e., the concentration of specific IgE antibodies to common ragweed pollen allergen, was determined by the REAST method and found to be 27.02±4.6 IU/mL (class 4). The specificity of binding of the PhRS IgE-Amb to common ragweed pollen allergen was confirmed. The results of tests for additional quality attributes (Description, Water, Mass homogeneity, Solubility, Tightness) met the requirements of the SP RF. The feasibility of using the PhRS IgE-Amb for determining the concentration of IgE antibodies in enterprise reference sera by direct ELISA and for assessing common ragweed pollen allergen activity by competitive ELISA was demonstrated. The coefficients of variation between the known values and those determined using the RS were 4.7% for direct ELISA and 8.0% for competitive ELISA.

CONCLUSIONS. For the first time, a Russian pharmacopoeial reference standard of lyophilized IgE-containing serum to common ragweed pollen allergen has been certified with the established characteristic — the concentration of specific IgE antibodies to common ragweed pollen allergen determined by REAST. The PhRS IgE-Amb complies with the requirements of the SP RF, is recommended for inclusion in the State Register of Reference Standards, and enables the standardization of allergens and reference sera in the Russian Federation.

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