分析および生物分析技術ジャーナル

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Validation of the Chromogenic Bioassay for the Potency Assessment of Streptokinase in Biopharmaceutical Formulations

Bruna Xavier, Raphael Leite Camponogara, Clóvis Dervil Appratto Cardoso Júnior, Rafaela Ferreira Perobelli, Mauricio Elesbão Walter, Fernanda Pavani Stamm Maldaner and Sérgio Luiz Dalmora

Streptokinase (STK) is a thrombolytic agent clinically used to treat patients with acute myocardial infarction and venous and arterial thrombosis. An in vitro chromogenic substrate end point bioassay was validated for the potency evaluation of biopharmaceutical formulations. The dose-response curve was linear over the concentration range of 2.50-40 IU/mL (r2=0.999), with a quantitation limit of 2.50 IU/mL and a detection limit of 1.10 IU/mL, respectively. Specificity was established in studies with spiked samples. The accuracy was 100.34% with bias lower than 0.53%, and method validation demonstrated also acceptable results for precision and robustness. The validated method was applied to the potency assessment giving potencies between 92.20% and 108.97%. In addition, the activity of streptodornase and streptolysin were also evaluated giving values lower than 9.79 IU per 100 000 IU STK, and 1.47 for the absorbance ratio, respectively. The validated bioassay was applied in combination with the purity evaluation, contributing to assure the batch-to-batch consistency and quality of the bulk and finished biotechnology-derived medicine.