Production and evaluation of a rabbit polyclonal antibody specific to the Aeromonas hydrophila outer membrane protein...

The outer membrane protein Omp38 of Aeromonas hydrophila, a major fish pathogen, is a potential candidate for vaccine development. In this study, we produce a rabbit polyclonal antibody specific to this protein. Recombinant Omp38, expressed in E. coli cells, was used as an antigen for rabbit immunization. The obtained antiserum successfully recognized Omp38 in Western blot, ELISA, and immunofluorescence staining. The antiserum could detect Omp38 at concentrations of ≥0.1563 ng well-1 and showed an endpoint titer of 1:5120000. Antibodies were further purified from antiserum using Protein A chromatography, yielding over 95% purity. The purified antibody could detect Omp38 at concentrations as low as 0.0391 ng well-1 at the 1:10000 dilution and retained reactivity at the 1:10240000 dilution. Overall, this study successfully generated a rabbit antiserum and a purified polyclonal antibody sample against Omp38, which can be applied in future studies related to this protein.