Preparation and identification of antiCK20 monoclonal antibody

Objective:To produce and identify antiCK20 monoclonal antibody.Methods:Lymphocytes from the spleen of mice being immunonized by CK20 antigen were fused with the myeloma cell line(SP2/0) using PEG4000.Hybridodma cells were established by selective growth of the fusion cells in the HT medium,and the presence of antiCK20 antibody was screened by inderect ELISA,and the clonality was achieved by limiting dilution.We have incubated cloning cells into mouse abdominal cavity to produce ascitic fluid contained monoclonal antibody.Chromatography with SPA-Sepharose CL-4B affinity column were emploied to isolate the monoclonal antibody from ascitic fluid.Finally,the antibody were tested the activities and sentivities,isoforms and titer through Western blot,two directions immuning diffusion of agar and ELISA.Results:Only one hybridoma cell line,secreating McAb against CK20,had been established.The modal number of chromosome is 101(99-103).The results of identifications showed that the antibodies kept high activitis and sensitivitis in detecting sample.The titer of ascitic fluid and the McAb purified are 1∶10~6 equally.The immunoglobulin of the McAb is classified as IgG1.Conclusion:AntiCK20 monoclonal antibody have been produced succesfully with high sensitive and active and was named L20031030.