Polymer-immobilized ready-to-use recombinant yeast assays for the detection of endocrine disruptive compounds

Scheme of recombinant yeast assays using three types of immobilizing matrices in a microplate or tubes. •Immobilization techniques applied to develop rapid ready-to-use assays.•Immobilized recombinant yeast used for detection of androgens and estrogens.•Recombinant yeast cells were immobilized in gelatin, Bacto agar and YPD agar.•Gelatin was the best immobilization matrix.•Immobilized yeast stored in fridge maintained sensitivity for at least 90d. Recombinant yeast assays (RYAs) constitute a suitable tool for the environmental monitoring of compounds with endocrine disrupting activities, notably estrogenicity and androgenicity. Conventional procedures require yeast reconstitution from frozen stock, which usually takes several days and demands additional equipment. With the aim of applying such assays to field studies and making them more accessible to less well-equipped laboratories, we have optimized RYA by the immobilization of Saccharomyces cerevisiae cells in three different polymer matrices – gelatin, Bacto agar, and Yeast Extract Peptone Dextrose agar – to obtain a ready-to-use version for the fast assessment of estrogenic and androgenic potencies of compounds and environmental samples. Among the three matrices, gelatin showed the best results for both testosterone (androgen receptor yeast strain; AR-RYA) and 17β-estradiol (estrogen receptor yeast strain; ER-RYA). AR-RYA was characterized by a lowest observed effect concentration (LOEC), EC50 and induction factor (IF) of 1nM, 2.2nM and 51, respectively. The values characterizing ER-RYA were 0.4nM, 1.8nM, and 63, respectively. Gelatin immobilization retained yeast viability and sensitivity for more than 90d of storage at 4°C. The use of the immobilized yeast reduced the assay duration to only 3h without necessity of sterile conditions. Because immobilized RYA can be performed either in multiwell microplates or glass tubes, it allows multiple samples to be tested at once, and easy adaptation to existing portable devices for direct in-field applications.