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(060) Development of quantitative vitellogenin-ELISA assays for fish test species used in endocrine disruptor screening. Nilsen, Bente1, Eidem, Janne1, Kristiansen, Sven1, Nilsen, Mona1, Berg, Karin1, Denslow, Nancy2, Goksøyr, Anders1,3, 1 2 3 ABSTRACT- Induction of the yolk protein precursor vitellogenin (Vtg) in plasma has proved to be a simple and sensitive biomarker for assessing exposure to environmental estrogens in fish. The widespread use of Vtg in this regard has lead to the need for standardized assays to quantify Vtg. Monoclonal antibodies, that can be produced from a single clone with a desired specificity and in unlimited amounts, have the potential to help accomplish this. Several governmental organisations, e.g. the OECD and EPA, have decided to incorporate standardized Vtg assays in screening programmes for endocrine disruptors. Using various combinations of monoclonal and polyclonal fish Vtg antibodies in a sandwich (capture) set-up, we have developed quantitative ELISA assays for rainbow trout (Oncorhynchus mykiss), and carp/fathead minnow (Cyprinus carpio/Pimephales promelas)Vtg. Similar assays for zebrafish (Danio rerio) and medaka (Oryzias latipes) are currently underway. The assays show a high sensitivity and are able to detect responses to estrogenic compounds such as 17b-estradiol and estrogen mimics in different test species. Key words: endocrine disrupters, vitellogenin, fish, ELISA |
