A new monoclonal antibody against vitellogenin from rainbow trout (Oncorhynchus mykiss)

Monoclonal antibodies were developed against vitellogenin (vtg) of rainbow trout, This protein is used as a biochemical response to the exposure with estrogenic compounds. Several mice were immunized with purified vtg and the spleen cells of these mice were fused with myeloma cells, The resulting hy bridoma cells were screened with an enzyme immunoassay for the production o f specific anti-vtg antibodies, Twelve positive cell lines were detected. T he hybridoma cell line B8D8 was adjusted to serum free medium; it produced monoclonal antibodies with a high selectivity and sensitivity. A detection limit of 5 mug/l vtg was achieved with a competitive enzyme immunoassay app lying a preincubation step and a streptavidin-biotin amplification system ( ABC system). Cross-reactivity with vtg of other species was detected with r oach (Rutilus rutilus), flounder (Platichthys flesus) and dab (Limanda lima nda). The assay was applied to rainbow trout (Oncorhynchus mykiss), which w ere exposed for six months to defined concentrations of effluents (10%, 20% , 30% and 40%) and surface water. Increased vtg levels in male fish are cor related with the increasing levels of effluent. The effluent levels in the exposure experiments are relevant for effluent loadings of the Berlin water ways during the seasons of the year. (C) 2001 Elsevier Science Ltd. All rig hts reserved.