Production of monoclonal antibodies against yellowtail ascites virus

Abstract

AbstractA total of 31 antibody‐secreting hybridoma cells against the yellowtail ascites virus (YAV) were established. These monoclonal antibodies (MAbs) reacted with the cytoplasm of YAV‐infected CHSE‐214 cells, but not with uninfected CHSE‐214 cells in an immunofluorescence test. Using these MAbs, two classes of polypeptides (VP2 and VP3) were characterized by immunoprecipitation followed by SDS‐PAGE, although one MAb did not react with either polypeptide. Fifteen out of the 17 MAbs that were reactive with VP2 polypeptides neutralized virus infectivity, but all 13 MAbs that were reactive with VP3 did not neutralize infectivity. In the immunofluorescence test, 29 out of the 31 MAbs obtained showed the same reaction pattern to 12 YAV isolates from yellowtail, Seriola quinqueradiata, goldstriped amberjack, Seriola aureovittata, and threeline grunt, Parapristipoma trilineatum, from different geographical regions. The remaining two MAbs showed slightly different reaction patterns to the YAV isolates. The reaction patterns of the MAbs to the VR‐299, Sp and Ab strains of IPNV were also investigated. Fourteen MAbs reacted to all three IPNV strains. The other 17 MAbs showed a negative reaction with at least one strain.