Development and characterization of polyclonal antibodies against subtype specific vitellogenin of the dojo loach, Misgurnus anguillicaudatus

Abstract Fish vitellogenin (Vtg) has become an important biomarker for assessing the estrogenic potency of chemicals and the exposure of animals to estrogenic contaminants present in aquatic environments. In the current study, the polyclonal antibodies against subtype-specific Vtgs of the dojo loach (Misgurnus anguillicaudatus) were developed and identified by gene recombination and expression. Immuno-biochemical analyses revealed that VtgAo1 protein appeared to be the major Vtg type in this species. Enhanced chemiluminescent Western blotting was developed using the antiserum against VtgAo1. Exposure of male loach to 17α-ethinylestradiol (EE2) and 17β-estradiol (E2) via water induced the VtgAo1 protein induction with a lowest-observed-effect concentration (LOEC) by 100 ng L−1 and 1000 ng L−1group at 7 day post-initiation (dpi), respectively. The results indicated that this method provided a valuable tool for detecting estrogenic activities in aquatic environments and excluded any difficulties expected in purification procedures to separate a highly pure Vtg subtype from circulating proteins, and the other Vtg subtypes.