PRODUCTION OF POLYCLONAL ANTIBODY AGAINST THE OUTER MEMBRANE PROTEIN Omp48 OF Aeromonas hydrophila

Abstract

Aeromonas hydrophila is one of the main pathogens in fish, causing a huge economic loss every year. Since outer membrane protein Omp48 had been demonstrated to be immunogenic in fish, it has gained attention as a potential vaccine candidate to protect fish from A. hydrophila.  In this study, we produced a polyclonal antibody against Omp48, which would be necessary for further studies on this protein. To reach this aim, 6xHis-tagged Omp48 was expressed in a recombinant E. coli strain and purified by a nickel affinity chromatography column, which yielded a purity of 97.3 ± 1.4%. The purified Omp48 was injected into rabbits in order to produce polyclonal anti-Omp48 antibody. Subsequently, we analyzed the obtained antiserum, which was demonstrated to have high specificity and sensitivity to the Omp48 protein by Western blot and ELISA, with the titer of 1:1,280,000. In addition, we also showed that the antiserum could be used to detect Omp48 in immunofluorescence staining.