Development, Purification, and Characterization of Igy Antibodies Against Vitellogenin of the "Vieja Colorada” Fish (Cichlasoma Festae), a Species Native to the Guayas River, Ecuador

Abstract

Vitellogenin (Vtg) is a specific phospho glycoprotein used as an environmental and steroidogenesis biomarker in fish. The present study shows the techniques to isolated IgY anti vitellogenin (Vtg) in Cichlasoma festae fish. Vtg was purified from adult males previously treated with estradiol and isolated from blood plasma using ion-exchange chromatography. Subsequently, Lohmann Brown hens were immunized at different doses, including three antigen booster doses. At day 45 post-immunization, posture eggs were collected, and successfully optimized the extraction of anti-Vtg -IgY antibody. The purified antibodies were validated with SDS-PAGE, detecting heavy and light chains of 70 kDa and 30 kDa, respectively. Further, we developed ELISA to estimate the Vtg specificity. Commercial anti-Vtg antibodies of Dorada (Sparus aurata) were used as a control, and immunoglobulins Y were validated for native species by Western Blot. The IgY anti-Vtg obtained by local conditions can be successfully used for programs in environmental monitoring and reproduction control in native aquatic species in Ecuador.