Development of a Sensitive Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) for Vitellogenin in the Japanese Eel Anguilla japonica

Abstract

A specific and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of low levels of serum vitellogenin (VTG) of the Japanese eel Anguilla japonica. The non-competitive avidin-biotm interaction was performed by a sandwich method using the specificity of the specific antiserum to eel yolk protein (a-lipovitellin) and biotinylated F(ab')2 fragment of a-lipovitellin. The assay could be run in 2 days and routinely detect VTG in a concentration as low as 0.8 ng/m/. The appearance of low levels of serum VTG significantly increased 9-12 h after a single injection of estradiol-17β into immature fish. The development of an ELISA for VTG made possible the quantification of serum VTG and thereby in vitro analysis of the mechanism of the onset and course of vitellogenesis. © 1995, The Japanese Society of Fisheries Science. All rights reserved.