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 disruptor
chemicals EDCs) through interfering with endocrine system, have the capability to induce developmental
and reproductive abnormalities in humans and animals. The yolk protein precursor vitellogenin (VTG) has
proved to be a simple and sensitive biomarker for assessing exposure of fish to EDCs especially the
estrogenic compounds. Work is ongoing to develop screening and testing programmes for endocrine
disrupting effects of new chemicals, and in the focus of this development are the fish test species common
carp (Cyprinous carpio). In this study we have developed quantitative enzyme-linked immunosorbent
assays (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 variation
of 3.9% (n = 10) and 12.5% (n = 30), respectively. This ELISA is capable of detecting VTG as low as 6
ng/ml, and can accurately detect VTG in even 10 μl of plasma. The ELISA was applied to measurement of
VTG production by male carp (Cyprinous carpio, Cyprinidae) fish exposed to ethynylestradiol. The results
showed that the amount of VTG produced in plasma of exposed fish increased in logarithmic order
comparing to the control group and the ELISA described here could be used as an indicator of water
pollution to estrogenic pollutants.