PARENT SESSION
1D Bioassays for specific hazards (estrogenic effects, genotoxicity, neurotoxicity, ...)
9:00 AM to 7:00 PM, Tuesday, 08 May 2001

(T/EH065) Monoclonal antibodies against zona radiata proteins for purification of individual Zrp-monomers and development of a quantitative Zrp-ELISA.

Goksøyr, Anders^1,2, Berg, Karin¹, Bringsvor, Kjersti², Nilsen, Mona¹, Walther, Bernt², Nilsen, Bente¹, ^{1 2}

ABSTRACT- An increasing number of chemicals, or their degradation products, have been found to possess estrogenic activity, leading to induction of the biomarker proteins vitellogenin (Vtg) and zona radiata proteins (Zrp) in fish. As a tool for monitoring the induction of Vtg in the OECD test species rainbow trout (Oncorhynchus mykiss), we have previously developed a quantitative rainbow trout Vtg ELISA. Some studies indicate that Zrp may be a more sensitive biomarker protein than Vtg. However, the different Zrp-monomers (3-4 proteins depending on species) has, to our knowledge, not yet been individually isolated and therefore not quantitatively analyzed. As a first step towards a quantitative assay for Zrp, we have produced monoclonal antibodies (MAbs) against Zrp with different affinities for -, -, and -Zrp from rainbow trout. Preliminary results indicate that these MAbs can be used for immunoaffinity purification of individual monomers. With purified Zrp-monomers and monomer-specific MAbs, quantitative ELISA-methods for individual Zrp-monomers in rainbow trout may be developed

Key words: endocrine disruptors, zona radiata proteins, ELISA, rainbow trout