Preparation And Identification Of Monoclonal Antibodies Against Caprine IFN-γ

IFN-γ arises by activation of T cells and NK cells.IFN-γ could strengthen the killing activity of macrophages and neutrophils,which have biological activities such as antiviral infection,anti-tumor,the change in concentrations of IFN-γ can be as an important index to assess the immune function at the inflammatory site and in blood.There has been no known report of the caprine IFN-γ monoclonal antibodies and the related kit so far.BALB/c mice were immunized with purified prokaryotic recombinant caprine IFN-γ in this study.For screening hybridoma cells secreting monoclonal antibodies against caprine IFN-γ,the technology of monoclonal antibody preparation was adopted,such as the conventional cell fusion.We carried out identification of the specificity and subtypes of IFN-γ monoclonal antibodies.The results were as follows:1.The titers of serum of three immunized BALB/c mice all could raise up to 105;2.After filtering of fused hybridoma cells,a strain of hybridoma cell named 3C was obtained,and the secreted monoclonal antibody could recognize IFN-γ.The titers of the selected strain of hybridoma cell supernatant、mice ascites were 103 and 106respectively.3.Monoclonal antibody against caprine IFN-γ was classified as IgG1,kappa type.4.Western blot analysis showed that,in this experiment,the monoclonal antibody could specifically recognize recombinant proteins of prokaryotic expression.5.The monoclonal antibody could react with IFN-γ in the PBMCs and could also react with the caprine IFN-γ which was produced by mastitis goat milk.The present research obtained a strain of hybridoma cell stably secreting monoclonal antibody recognizing caprine IFN-γ,and the production of monoclonal antibody secreted by hybridoma cell was high.