Padronização e Validação da técnica do Limulus Amebocyte Lysate (LAL) Semi-Quantitativa e Quantitativa para o Biofármaco Alfainterferona 2b Humana Recombinante

Abstract

In recent years, the Pharmacopean, technical reports and international guides for pharmaceuticals products requires each time more in their monographies the application of Limulus Ambocytes Lysate endotoxins assay (LAL) for release and pyrogen control in parenteral finished products. The objective of this study was the standardization and validation of the LAL test for the human recombinant biopharmaceutical  interferon alpha 2b, produced in E. coli, were used Gel-Clot and Chromogenics Kinetic methods (semi-quantitative and quantitative). In the case of Chromogenics method test, the maximum valid dilution (MDV) was calculated for each presentation based on the concentration of product (3 MUI, MDV: 1:3,888 and 10 MUI, MVD: 1:12,962), serial dilutions under the MDV (1:80) were evaluated in triplicate to detect interferences. For the gel-clot test for the 3 MUI presentation (MDV: 1:17) 1:8 dilution was set for interferences detection test. For tests’ validation, several dilutions were performed using standard endotoxin concentrations in 0,005 to 50 EU / ml to confirm the criteria for the methods performance (linearity, specificity, accuracy, reply, reproducibility). The results of validation showed that all pharmacopeia and regulatory parameters (ANVISA) studied, are compatible with the MDV used for each studied presentation of the human recombinant biopharmaceuticals  interferon alpha 2b and may be used in quality control.