Preparation and identification of a monoclonal antibody against FLT3 receptor

Objective To prepare and identify the monoclonal ntibody(mAb) against FLT3 receptor.Methods BALB/C mice were immunized with protein from the spleen cells isolated and fused with SP2/0 cells.After several rounds of screening and cloning,the hybridoma cell strain secreting anti-FLT3 mAb was obtained.Its specificity was evaluated with ELISA and Western blot,and the titer,immunoglobulin subtype and affinity of the mAb were measured.Results Two cell strains of hybridoma were obtained and named as 5F3B11B1 and D3H2C12.The hybridoma cell strain secreting anti-FLT3 receptor mAb was identified to belong to IgG1 subtype.The mAb titers in ascetic fluid were 1:409600 and 1:102400,as determined with ELISA with affinity reaching 7.85×106 L/mol.Western blot demonstrated that the antibodies could specifically recognize the immunogen.The cell immunohistochemistry proved that the antibody could recognize the FLT3 receptor antigens expressed on the blast cells in acute myelogenous leukemia.Conclusion The success in anti-FLT3 receptor mAb preparation provides the basis for further study of hematologic disease such as leukemia.