Vitellogenin in Zoarces viviparus: Purification, quantification by ELISA and induction by estradiol-17 β and 4-nonylphenol

Abstract

Vitellogenin was isolated by gel filtration and ion-exchange chromatography from plasma of estradiol-treated male Zoarces viviparus. The purification of vitellogenin was followed through each step by native polyacrylamide gelelectrophoresis (PAGE). The purified vitellogenin was used to raise anti-vitellogenin antibodies in rabbits. The specificity of the affinity purified antibody raised against vitellogenin was assessed by Western blot analysis. No crossreactivity was observed with plasma from non-induced control males. A direct enzyme-linked immunosorbent assay (ELISA) was developed by use of the raised anti-vitellogenin. The detection limit for the purified standard vitellogenin by the ELISA method was 5 ng ml −1 Vitellogenin levels were quantified in plasma of pregnant female Zoarces viviparus, which were held in aquaria with different concentrations of the estrogenic compound 4-nonylphenol dissolved in the ambient seawater. A marked effect of exposure to ambient 4-nonylphenol was observed by significant dose-dependent increases in the level of plasma vitellogenin of the pregnant fish as well as of embryos exposed in vitro.