A New Approach Method to Study Thyroid Hormone Disruption: Optimization and Standardization of an Assay to Assess the Inhibition of DIO1 Enzyme in Human Liver Microsomes
Here, the DIO1-SK assay using human liver microsomes as a DIO1 enzyme source and the Sandell-Kolthoff (SK) reaction as a colorimetric readout to quantify released iodide was standardized and optimized according to Good In Vitro Method Practices (GIVIMP). Potential pitfalls are described, and possible solutions are provided. The reproducibility of the method was demonstrated by testing a set of six described DIO1 inhibitors in five independent assay runs. The results of this study were used to amend the testing protocol: a set of acceptance criteria was defined to assess the validity of assay runs and strategies to verify the specificity of observed DIO1 interaction were implemented.
WEBER Andreas Georg;
BIRK Barbara;
HERRMANN Chantal;
HUENER Hans-Albrecht;
RENKO Kostja;
COECKE Sandra;
SCHNEIDER Steffen;
FUNK-WEYER Dorothee;
LANDSIEDEL Robert;
2024-01-15
Mary Ann Liebert, Inc.
JRC135690
2332-1512 (online),
https://www.liebertpub.com/doi/full/10.1089/aivt.2022.0010,
https://doi.org/10.1089/aivt.2022.0010,
https://publications.jrc.ec.europa.eu/repository/handle/JRC135690,
10.1089/aivt.2022.0010 (online),
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