First report on the isolation of immunoglobulin M of guppy, Poecilia reticulata, for the production of polyclonal antibodies

Abstract

As guppies are one of the commercially important fish in freshwater ornamental aquaculture industry, it is important to gain an understanding of guppy immune response for infectious disease control. Till now, the number of study that examined the immune response of guppies is limited and effective tools for monitoring guppy antibody have not been reported. In this study, we successfully isolated guppy IgM using mannan-binding protein (MBP) affinity chromatography and produced specific polyclonal antibodies against guppy IgM heavy and light chains, that showed a molecular weight of approximately 74 and 23 kDa respectively. The produced polyclonal antibodies were used to develop an enzyme-linked immunosorbent assay (ELISA) and have demonstrated to be an effective tool for the detection and quantification of antigen-specific antibody of guppies immunized with Pseudomonas fluorescens. In conclusion, the produced anti-guppy IgM polyclonal antibodies should prove its future implications for immunology and epidemiology studies in guppies. © 2009 Elsevier B.V. All rights reserved.