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Retrofitting an Estrogen Receptor Transactivation Assay with Metabolic Competence Using Alginate Immobilization of Metabolic Enzymes (AIME) (OpenTox)
Citation:
DeGroot, D., R. Thomas, P. Carmichael, M. Lee, AND C. Deisenroth. Retrofitting an Estrogen Receptor Transactivation Assay with Metabolic Competence Using Alginate Immobilization of Metabolic Enzymes (AIME) (OpenTox). Presented at OpenTox, RTP, NC, July 11 - 12, 2018. https://doi.org/10.23645/epacomptox.7056323
Impact/Purpose:
The Alginate Immobilization of Metabolic Enzymes (AIME) platform is an HTS-compatible solution that retrofits existing assays with metabolic competence by attaching alginate-hepatic S9 microspheres to solid supports extending from microplate lids. This work supports US EPA efforts to enhance evaluation of potential chemical hazards by incorporating xenobiotic metabolism into existing high-throughput assays. Poster presentation at Open Tox July 2018.
Description:
The VM7Luc4E2 estrogen receptor (ER) transactivation assay is an OECD approved method (TG 457) for the detection of ER agonists and antagonists, and is also part of the Tox21 high-throughput screening (HTS) portfolio. Despite international acceptance as a screening assay, immortalized cell lines such as VM7Luc4E2 do not express a full complement of metabolizing enzymes. This has led to calls for improved methods for the incorporation of metabolic competence into in vitro assays, particularly those used in the detection of endocrine active chemicals. The Alginate Immobilization of Metabolic Enzymes (AIME) platform is an HTS-compatible solution that retrofits existing assays with metabolic competence by attaching alginate-hepatic S9 microspheres to solid supports extending from microplate lids. Herein we demonstrate the successful retrofit of the VM7Luc4E2 ER transactivation assay with metabolic competence using the AIME platform and methoxychlor as a proof-of-concept reference compound. This work supports US EPA efforts to enhance evaluation of potential chemical hazards by incorporating xenobiotic metabolism into existing high-throughput assays. This abstract does not necessarily reflect the policy of the US EPA.
URLs/Downloads:
DOI: Retrofitting an Estrogen Receptor Transactivation Assay with Metabolic Competence Using Alginate Immobilization of Metabolic Enzymes (AIME) (OpenTox)
DEGROOT_DEISENROTH_OPENTOX.PDF (PDF, NA pp, 299.775 KB, about PDF)