[Preparation and characterization of monoclonal antibodies against F domain of human progranulin].

OBJECTIVE: To prepare and characterize the monoclonal antibody (mAb) against F domain of human progranulin (GrnF). METHODS: Yeast expression vector containing GrnF gene was constructed using molecular biological technology. Then eukaryotic fusion protein human serum albumin (HSA)-GrnF from yeast vector was expressed and purified. BALB/c mice were immunized by the purified HSA-GrnF fusion protein. The splenocytes of the BALB/c mice were isolated from spleen and fused with Sp2/0 myeloma cells. Indirect ELISA and limiting dilution assay were used for screening hybridoma cell lines. The specificity of monoclonal antibodies against GrnF was evaluated with indirect ELISA, Western blotting and immunohistochemistry. The immunoglobulin subclass was identified with mouse monoclonal antibody isotyping reagents. RESULTS: Two hybridoma cell lines, 1G6 and 4E8, were obtained and heavy chain subclasses of the two hybridoma cell lines were IgG1. CONCLUSION: The mAbs that can specifically recognize GrnF have been successfully prepared.