Generating a monoclonal antibody against chicken interferon gamma for the establishment of quantitative ELISA

BALB/c mice were immunized four times with recombinant chicken interferon-gamma expressed in E.coli at 100μg per mice and cell fusion was conducted according to standard procedure.Positive hybridoma clones were screened by indirect ELISA.A hybridoma that stably secreted specific monoclonal antibody against chicken gamma interferon was developed. The monoclonal antibody,named G1 strain belonged to IgG1 isotype and hadκchain.Western blot showed that G1 strain reacted strongly with recombinant chicken interferon-gamma expressed either in baculovirus expression systerm or in E.coli.However the recombinant chicken interferon-gamma could not inhibit the replication of the VSV~*GFP in CEF cells.Relative quantitative ELISA was successfully established using the monoclonal antibody and recombinant chicken interferon-gamma expressed in sf9 cell.