Preparation and identification of the monoclonal antibody against human PSMA7 antigen / 解放军医学杂志

ABSTRACT

Objective To prepare the PSMA7 hybridoma cell lines using classical hybridoma technique and to perform purification and elementary identification of the monoclonal antibodies(McAbs)against PSMA7 antigen for further study.Methods BALB/c mice were immunized with purified recombinant PSMA7 protein.Once the antibody titer of blood taken from mouse tail reached 1∶6 400,a cell fusion was performed between mouse splenic cells and myeloma cells(Sp2/0),and then the hybridoma cell lines secreting monoclonal antibodies against PSMA7 antigen were screened by indirect enzyme linked immunosorbent assay(ELISA).The immunoglobulin subtypes and titer of the monoclonal antibodies against PSMA7 antigen were identified and measured by Western blot analysis and enzyme linked immunosorbent assay,respectively.Results After cell fusion and subcloning,two hybridoma cell lines that stably secreted monoclonal antibodies against PSMA7 antigen were successfully obtained:B013 and B001,which belong to the subtypes of IgG1.The antibody titers in the hybridoma culture supernatant were 1∶128 and 1∶256,and those in the ascites fluid were 1∶25 600 and 1∶32 000,respectively.Western blot analysis and immunohistochemistry showed that the two antibodies can specifically bind with PSMA7 antigen derived from human eucaryotic cells or tissue.Conclusions Two monoclonal antibodies against PSMA7 antigen with high titers and specificity have been successfully prepared.These antibodies can be used for the identification of PSMA7 protein,and may be a useful tool for studying the biological properties of PSMA7 protein.