Determining and Estimation of Antibody Production in the Bubble Eye, Goldfish

Development of antibody production technologies is necessary for diagnostic treatments and drug discovery. In general, mammals are used as host animals to produce antigen-specific antibody. However, such host animals have never produced several specific antibodies because host animals may not recognize foreign proteins. To overcome this problem, we used teleost to produce antibodies because teleost are evolutionally localized in the origin of vertebrates and have an acquired immune system in addition to the innate immune system. In particular, we attempt to produce antibody using “Bubble Eye” as a kind of goldfish (Carassius auratus), which has sacs filled with lymph liquid, as an immune animal. In this study, a recombinant EGFP-His was expressed in E. coli and then injected into Bubble Eye’s sac in every two weeks. The antibodies were collected from sac instead of blood. Furthermore, a sandwich dot blotting was developed for detection of antibodies against EGFP-His. The antigen-specific antibodies were detected after 42 days from first immunization.