Freeze-drying as suitable method to achieve ready-to-use yeast biosensors for androgenic and estrogenic compounds
| Authors | |
|---|---|
| Year of publication | 2016 |
| Type | Article in Periodical |
| Magazine / Source | Chemosphere |
| MU Faculty or unit | |
| Citation | |
| web | http://www.sciencedirect.com/science/article/pii/S0045653516300376 |
| Doi | http://dx.doi.org/10.1016/j.chemosphere.2016.01.038 |
| Field | Environment influence on health |
| Keywords | Recombinant yeast assay; Freeze-drying; Estrogen; Androgen; Long-term; Ready-to-use |
| Description | Recombinant yeast assays (RYAS) have been proved to be a suitable tool for the fast screening of compounds with endocrine disrupting activities. However, ready-to-use versions more accessible to less equipped laboratories and field studies are scarce and far from optimal throughputs. Here, we have applied freeze-drying technology to optimize RYA for the fast assessment of environmental compounds with estrogenic and androgenic potencies. The effects of different cryoprotectants, initial optical density and long-term storage were evaluated. The study included detailed characterization of sensitivity, robustness and reproducibility of the new ready-to-use versions, as well as comparison with the standard assays. Freeze-dried RYAS showed similar dose-responses curves to their homolog standard assays, with Lowest Observed Effect Concentration (LOEC) and Median effective Concentration (EC50) of 1 nM and 7.5 nM for testosterone, and 0.05 nM and 0.5 nM for 17 beta-estradiol, respectively. Freeze-dried cells stored at 4 degrees C retained maximum sensitivity up to 2 months, while cells stored at -18 degrees C showed no decrease in sensitivity throughout the study (10 months). This ready-to-use RYA is easily accessible and may be potentially used for on-site applications. |
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