Lipovitellin as an antigen to improve the precision of sandwich ELISA for quantifying zebrafish (Danio rerio) vitellogenin.

Vitellogenin (Vtg) in zebrafish (Danio rerio) is a core biomarker for screening environmental estrogens in test guidelines of the Organization for Economic Cooperation and Development. To accurately quantify zebrafish Vtg, lipovitellin (Lv), the main Vtg-derived yolk protein, was used as the antigen to establish a sandwich enzyme-linked immunosorbent assay (ELISA). The purified Lv was a phospholipoglycoprotein with apparent molecular weight of ~445kDa, and separated into three polypeptides corresponding to ~117, ~102, and ~23.

Juvenile zebrafish in the vitellogenin blank period as an alternative test organism for evaluation of estrogenic activity of chemicals.

The present study aimed to determine the suitable development period for zebrafish to evaluate estrogenic activities accurately. An enzyme-linked immunosorbent assay was developed and used to detect the vitellogenin (Vtg)-derived yolk proteins and newly produced Vtg, and 9 d to 56 d posthatching was determined as the Vtg-blank period. Juveniles in this period were found to have lower baseline Vtg levels than adult males and were considered an alternative test organism for detecting environmental estrogens.

Development of a lipovitellin-based sandwich ELISA for quantification of vitellogenin in surface mucus and plasma of goldfish (Carassius auratus).

Goldfish (Carassius auratus) vitellogenin (Vtg) is an efficient biomarker for estrogen contamination in aquatic environments. In this study, Vtg and lipovitellin (Lv) were purified from the plasma of 17β-estradiol (E2)-induced male goldfish and unfertilized eggs of females, and were used to generate polyclonal antibodies against Vtg (anti-Vtg) and Lv (anti-Lv), respectively. SDS-PAGE and Western blot were performed to confirm the specificity of the two antibodies and the immunological similarity between Vtg and Lv.