Vitellogenin assay by enzyme-linked immunosorbent assay (ELISA) as a biomarker of endocrine disruptor chemicals (EDCs) pollution

There are increasing evidences that show many xenobiotic chemicals (called as endocrine disruptorchemicals EDCs) through interfering with endocrine system, have the capability to induce developmentaland reproductive abnormalities in humans and animals. The yolk protein precursor vitellogenin (VTG) hasproved to be a simple and sensitive biomarker for assessing exposure of fish to EDCs especially theestrogenic compounds. Work is ongoing to develop screening and testing programmes for endocrinedisrupting effects of new chemicals, and in the focus of this development are the fish test species commoncarp (Cyprinous carpio). In this study we have developed quantitative enzyme-linked immunosorbentassays (ELISA) for VTG in common carp. The working range of the ELISA was 11.25 to 2,000 ng/ml (25-75% specific binding/maximum antibody binding [B/B0]) with a 50% B/B0 intra- and interassay variationof 3.9% (n = 10) and 12.5% (n = 30), respectively. This ELISA is capable of detecting VTG as low as 6ng/ml, and can accurately detect VTG in even 10 μl of plasma. The ELISA was applied to measurement ofVTG production by male carp (Cyprinous carpio, Cyprinidae) fish exposed to ethynylestradiol. The resultsshowed that the amount of VTG produced in plasma of exposed fish increased in logarithmic ordercomparing to the control group and the ELISA described here could be used as an indicator of waterpollution to estrogenic pollutants.