Enzyme-linked immunosorbent assay (ELISA) for brown trout vitellogenin for use in the detection of environmental estrogens

Conference · Sun Dec 31 04:00:00 EST 1995

OSTI ID:218432

Gamble, A; Solomon, K ^[1]; Sherry, J; Fielden, M; Hodson, P ^[2]

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1. Univ. of Guelph, Ontario (Canada)
2. Environment Canada, Budington, Ontario (Canada)

Vitellogenin (Vg) is a phospholipoprotein egg yolk precursor found in females of most oviparous vertebrates. Vitellogenin induction in male teleosts is known to result from exposure to xenoestrogens. Using antisera for brown trout Vg, the authors adapted an (ELISA) for in vivo Vg in brown trout (Salmo trutta). The authors will use the Vg bioassay to detect the estrogenic effects of effluents, contaminated sediments and suspect chemicals. Vg production in male fish was induced by intraperitoneal injection of 11-B estradiol. Harvested and purified Vg was used in an assay based on competition between soluble Vg and Vg adsorbed in microtitre wells for binding sites on rabbit anti-Vg antibody. Adsorbed Vg-antibody complexes were subsequently revealed by an alkaline phosphatase technique. The reaction intensity was measured as absorbance and used to quantify the amount of antibody bound to adsorbed Vg. The amount of bound antibody was inversely proportional to the amount of vitellogenin in the fish plasma. Preliminary results show that the assay`s working range, corresponding to 80--20% binding, was 14--355 ng/ml, with 50% binding (IC{sub 50}) around 71 ng/mi. Calculated at 50% binding, the intra-assay variation was less than 8% (n = 9) and the inter-assay variation was also less than 8% (n =4). The assay`s sensitivity was 0.067 ng/mi and the limit of detection was 0.134 ng/ml. The authors will present the results of Vg bioassays for the presence of estrogen mimics in industrial effluent.

• OSTI ID: 218432
• Report Number(s): CONF-9511137--; ISBN 1-880611-03-1
• Country of Publication: United States
• Language: English

Related Subjects

54 ENVIRONMENTAL SCIENCES
56 BIOLOGY AND MEDICINE
APPLIED STUDIES
BIOLOGICAL EFFECTS
BIOLOGICAL INDICATORS
BIOLOGICAL MARKERS
BIOLOGICAL STRESS
CHEMICAL EFFLUENTS
ENZYME IMMUNOASSAY
EVALUATION
LIPOPROTEINS
PHOSPHOPROTEINS
TROUT
WATER POLLUTION