Resumo

RAMIREZ GONZALEZ, Wendy et al. Interlaboratory validation of the SDS-PAGE method for the protein composition determination in house dust mite allergen extracts. Rev. CENIC Cienc. Biol [online]. 2022, vol.53, n.1, pp. 1-13.  Epub 07-Mar-2022. ISSN 2221-2450.

The European Pharmacopoeia Monograph on Allergen Products establishes the importance of protein composition determination to final products and also to the Allergenic Raw Material (ARM) and the Active Pharmaceutical Ingredient (API). Analytical methods validation ensures the quality of the product from its initial stages. The aim of this work was to validate SDS-PAGE method for allergenic extracts of Dermatophagoides siboney (Ds) and Blomia tropicalis (Bt) mites in the three stages of the process (ARM, API and final product). Three BIOCEN laboratories participated on this study. Precision (Repeatability, Intermediate Precision and Reproducibility) was evaluated in relative content determination of the main bands that contain allergenic components. As a Specificity parameter of the method, the possible interference of ammonium bicarbonate was analyzed. Accuracy was evaluated and the Limit of Detection (LD) was determined in the molecular weight (MW) determination of the main bands. All established acceptance criteria were met. The method was precise, all the coefficients of variation (CV) calculated were <5% and there were no significant differences between the CVs compared by Fisher's test, p>0.05. There were no significant differences between percentages of the main bands in the allergenic extracts with Ammonium Bicarbonate in its original composition according to ANOVA test, p>0.05. The method was exact, there were no significant differences (t Student test, p> 0.05) between the MW experimental values and those reported in the literature. The LD was for MPA of Ds 5000UB, Bt 50000UB; IFA and Final Product of Ds 2500UB, Bt 12500UB. In conclusion, the SDS-PAGE analytical technique was validated for the protein composition determination in the house dust mite allergen extracts, in three different stages of the manufacturing process. Thus, the quality assurance of these products is strengthened.

Palavras-chave : Advanced Education; pharmaceutical product; performance testing.

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