PT07 Chemical and Biological Analysis of Endocrine Disrupting Compounds
Exhibit Hall
8:00 AM - Tuesday
(PT111) Development and implementation of an enzyme-linked immunosorbant assay for the measurement of vitellogenin in flatfish from the Pacific Ocean.
Irwin, M.A.1, Armstrong, J.2, Sakamoto, K.2, Schlenk, D.1, 1 UC Riverside, Riverside, CA, U.S.A.2 Orange County Sanitation District, Fountain Valley, CA, U.S.A.
ABSTRACT- The presence of chemicals in the environment that induce estrogenic effects has become an increasing concern in southern California. Flatfish are particularly susceptible to these compounds as they are intimately associated with sediments to which the chemicals can adsorb, and the fish can be found in close proximity to outfalls where concentrations of these compounds may be high. Development of quantitative assays to measure estrogenic effects in flatfish off southern California is of great importance to determine the degree and extent of these effects in the native population. To this end this study was conducted to develop a competitive binding, absorbance-based enzyme-linked immunosorbant assay (ELISA) with which to measure concentrations of vitellogenin in flatfish. Vitellogenin, an egg-yolk precursor protein, is a commonly used biomarker of estrogenic exposure in fish. The assay was developed to measure vitellogenin in hornyhead turbot (Pleuronichthys verticalis), English sole (Pleuronectes [or Parophrys] vetulus), and California halibut (Paralichthys californicus). The assay used California halibut vitellogenin as the standard, rabbit anti-turbot vitellogenin as the primary antibody, and goat anti-rabbit labeled with alkaline phosphatase as the secondary antibody. The working range of the ELISA was 100 to 3200 ng/ml of vitellogenin. Comparison of standard curves indicated that results of multiple tests could be compared. The ELISA could detect an estrogenic dose response. The ELISA appeared to be adequate for measurement of hornyhead turbot and California halibut vitellogenin, but there appeared to be interferences with measurement of English sole vitellogenin. This method was used in measurement of estrogenic effects on flatfish in an effluent monitoring program off southern California.
Key words: vitellogenin, endocrine disruption, flatfish, ELISA