2001 Volume 4 Issue 2 Pages 87-98
An enzyme-linked immunosorbent assay (ELISA) was developed to quantify hepatic vitellogenin (VTG) in medaka (Oryzias latipes). VTG quantification was performed by a direct sandwich method using specific monoclonal antibodies to medaka VTG and biotinylated polyclonal antibodies. The ELISA's performance was optimized and characterized. The assay's working range was 0.488 to 500 ng/mL; intra- and inter-assay variations were 3.0% to 7.1% (n=4) and 0.7% to 8.3% (n=3), respectively. Recovery and extraction studies with liver homogenate samples showed that this assay is well applicable to the determination of the hepatic VTG in medaka. An exposure study with 17β-estradiol demonstrated that this ELISA assay was available to quantify the VTG in the livers of individual medaka. These results indicate that the ELISA assay with liver homogenate samples is suitable for use as an in vivo screening assay for determination of the hepatic VTG in medaka exposed to xcno-estrogens.
