Validation of a cell-based colorimetric reporter gene assay for the evaluation of Type I Interferons

•The HEK-Blue IFN-α/β system was validated to comply international guidelines.•This system complied in accuracy, precision, specificity and, system suitability.•The validated system showed robustness to evaluate different commercial type I IFN.•This bioassay could be a complementary method to evaluate type I IFN activity.•The system met the validation criteria to be used in pharmaceutical environments. The biotherapeutic type I interferons (IFN-I) are indicated to treat several diseases. These products are regulated to guarantee safety and efficacy through critical quality attributes. For this purpose, the development of robust assays is required, followed by its validation to demonstrate their suitability for its intended purpose. Despite there are some commercial kits to evaluate IFN-I signaling, these are focused on measuring in vitro biological response instead of their validation, which is a pharmaceutical industry requirement. The aim of this work was to validate the HEK-Blue IFN-α/β system evaluating the biological activity of IFN-α/β under good laboratory practices, according to international standards. Our results demonstrated that HEK-Blue IFN-α/β system comply with accuracy (r2>0.95) precision (CV