A sensitive recombinant cell-based bioluminescent assay for detection of androgen-like compounds

Elisa Michelini, Luca Cevenini, Laura Mezzanotte, Piia Leskinen, Marko Virta, Matti Karp, Aldo Roda

Research output: Contribution to journalArticleScientificpeer-review

Abstract

We report a step-by-step protocol describing how to develop and use a yeast-based bioassay for androgen-like compounds. Saccharomyces cerevisiae cells are genetically engineered to express the human androgen receptor (hAR) and the bioluminescent (BL) reporter gene luciferase (from Photinus pyralis) under the control of the androgen response element (ARE). In the presence of androgens, activated hAR binds to the ARE sequences and activates luciferase expression. After addition of D-luciferin, luciferase activity measurements can be performed, and the BL signal is proportional to the androgenic activity of the sample. Cytotoxic effects of the sample are monitored by the use of a control yeast strain that allows BL signal correction according to cell viability. After overnight culture of the recombinant strain, the assay can be accomplished in a 96-well microplate format in 1 working day with a detection limit of 0.05 nM for testosterone and intra-and interassay variability of 14% and 23%, respectively.
Original languageEnglish
JournalNature Protocols
Volume3
Issue number12
Pages (from-to)1895 - 1902
ISSN1754-2189
DOIs
Publication statusPublished - 2008
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 1182 Biochemistry, cell and molecular biology

Projects

Bioavailability of metals and xenobiotics

Hynninen, A., Rajasärkkä, J., Virta, M. & Muurinen, J.

01/01/2001 → …

Project: Research project

Cite this

Michelini, E., Cevenini, L., Mezzanotte, L., Leskinen, P., Virta, M., Karp, M., & Roda, A. (2008). A sensitive recombinant cell-based bioluminescent assay for detection of androgen-like compounds. Nature Protocols, 3(12), 1895 - 1902. https://doi.org/10.1038/nprot.2008.189