Evaluation of monoclonal antibody technology for the species-specific identification of Centropomus undecimalis

A fusion was conducted using spleen cells from a hyperimmunized Balb/C mouse and a murine myeloma cell line, in an attempt to create monoclonal antibodies specific to parvalbumin from Centropomus undecimalis. The fusion protocol utilized a 24 well culture plate protocol. Initial results indicated the successful generation of antibody producing hybridomas specific to the desired parvalbumin; however, attempts to isolate and clone these cells were unsuccessful. The study results clearly demonstrate some potential drawbacks to using the 24-well fusion protocol versus a 96-well protocol. The 24-well protocol seems to favor rapidly growing non-secreting cells due to the lower dilutions involved. This study was successful in isolating murine polyclonal antiserum reactive in isolating murine polyclonal antiserum reactive with piscine parvalbumin. Further, initial cultures of hybridomas were able to distinguish C. undecimalis from several other species of fishes thus confirming the suitability of parvalbumin as a biomaker protein.