Preparation and iIdentification of murine monoclonal antibody against EPF

Objective To prepare and identify the murine monoclonal antibody(McAb)against early pregnancy factor(EPF).Methods BALB/c mice were immunized with recombinant EPF,based on which the McAb was prepared by hybridoma technique,then purified and determined for titer by indirect ELISA,for antigenic epitopes by additive ELISA,for specificity by Western blotting,and analyzed for class,subclass and chromosome karyotypes.Results Two cell strains secreting McAb against EPF were screened.The McAbs showed high activity.Both two McAbs were IgG1 with κ chain,and their titers were higher than 2×10-5 in ascites.The chromosome karyotypes of the 2 cell strains were consistent with those of hybridoma cells.The antigenic epitopes of two McAbs were different.McAbs have high purity and specificity detected by SDS-PAGE and Western blotting respectively.Conclusion The murine McAbs against EPF with high activity were successfully prepared.