The preparation and characterization of monoclonal antibodies against phosphorylated-tyrosine in intracellular region of human CD130

Aim To obtain the tyrosine-phosphorylated monoclonal antibodies(mAbs)against human CD130 molecules. Methods The peptide(N752-G766)of CD130 was used as the antigen. Hybridoma cell lines were established by cell fusion. The mAbs were characterized by ELISA,Western blot, immunoprecipitation and immunohistochemical staining. Results (1)4 hybridoma cell lines(MIY1,MIY2,MIY3 and MIY4) could secrete stably mAbs which can react specificly to N752-G766 peptide.(2) The results of Western blot showed that mAbs could recognize molecules of relative molecular mass(Mr) 13×104～14×104 in the target cells. The mAbs could immunoprecipitate molecules recognized by anti-CD130 mAb.(3)The molecules of Mr being 13×104～14×104 recognized by the mAbs ro anti CD130 mAb were the intracellular tyrosine-phosphorylated proteins induced by IL-6. Conclusion Antibodies against phosphorylated tyrosine in human CD130 molecules has been prepared and characterized successfully.