[Prepared the monoclonal antibody associated with hepatocellular carcinoma and analyzed its epitope].

Aim: To prepare the monoclonal antibodies associated with hepatocellular carcinoma (HCC) for diagnosis.
Methods: 3 BALB/c mice were immunized with high metastasis characteristic HCC cells (HCCLM-6). The splenocytes from the immunized mice were fused with murine myeloma cells (Sp2/0). The hybridoma cells were screened by ELISA after coating with the total protein and the culture supernatant of HCCLM-6 cells. The mAbs were characterized by immunohistochemical staining in the HCC tissues, and by indirect immunofIuorescent staining in different cell lines. The antigen and epitope recognized by the mAbs were identified by the screening premade Uni-ZAP human liver cDNA expression library.
Results: Twenty-eight hybridoma cells secreting mAbs were established. One clone of the hybridomas, QGA062, secreted specific mAb associated with HCC. The antigen recognized by the mAb QGA062 was identified as fibronectin (FN), and the epitope was localized among the peptide YTVSLVAIKGNQESPK.
Conclusion: The mAb against a HCC-associated epitope in FN is established and characterized, will be a very useful reagent for diagnosis of HCC.