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

Alexander Marx, Jim Sherry, Peter Dieter Hansen, Bertold Hock

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

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 hybridoma cells were screened with an enzyme immunoassay for the production of specific anti-vtg antibodies. Twelve positive cell lines were detected. The 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 μg/l vtg was achieved with a competitive enzyme immunoassay applying a preincubation step and a streptavidin-biotin amplification system (ABC system). Cross-reactivity with vtg of other species was detected with roach (Rutilus rutilus), flounder (Platichthysflesus) and dab (Limanda limanda). The assay was applied to rainbow trout (Oncorhynchus mykiss), which were exposed for six months to defined concentrations of effluents (10%, 20%, 30% and 40%) and surface water. Increased vtg levels in male fish are correlated with the increasing levels of effluent. The effluent levels in the exposure experiments are relevant for effluent loadings of the Berlin waterways during the seasons of the year.

Original languageEnglish
Pages (from-to)393-399
Number of pages7
JournalChemosphere
Volume44
Issue number3
DOIs
StatePublished - 2001

Keywords

  • Endocrine disruptors
  • Enzyme immunoassay
  • Monoclonal antibodies
  • Rainbow trout
  • Vitellogenin

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