Vitellogenin assay by enzyme-linked immunosorbant assay as a biomarker of endocrine disruptor chemicals pollution

Abstract

There is increasing evidence that many xenobiotic chemicals (called as endocrine disruptor chemicals, EDCs) through interfering with the 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 (Cyprinus carpio). In this study, we have developed quantitative enzyme-linked immunosorbent assays (ELISA) for VTS in common carp. The working range of the ELISA was 11.25 -2000 ng/ml (75 -25% 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 ml of plasma. The ELISA was applied to measurement of Vtg production by male carp (C. carpio, Cyprinidae) fish exposure 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.

Biomarker Genes for Detecting Estrogenic Activity of Endocrine Disruptors via Estrogen Receptors

“…Male fish also carry the VTG gene, although VTG protein is normally not expressed because the circulation levels of E2 are extremely low in male blood plasma [30]. However, males still have the capability to express VTG, and male fish are known to produce the protein under the influence of estrogenic EDs [30,31,32]. E2, NP, and OP all induce the expression of VTG in male fish in a dose-dependent manner [33,34], suggesting that the VTG gene in male fish can be used as a biomarker for evaluating the effects of EDs [35,36,37].…”

Selection and characterisation of single-stranded DNA aptamers for triclosan

“…In addition, these methods typically require sophisticated laboratory-based environment and highly trained personnel, and are not really amendable for decentralised analyses. Alternatively, conventional receptor-based assays, such as immunoassays have been developed with some success for the analysis of some EDCs (Ebrahimi 2007;King and Hassell 2008;Cha et al 2012). These assays utilise antibodies as molecular recognition elements, and are more cost-effective compared with chromatographic techniques, but lag in terms of stability, production time and often specificity.…”