| [AIM] The primary carcinoma of liver (PCL) is the 6th most commen cancer in the world and its incidence in China is much higher. The total number of liver carcinoma patients in the world in 2000 is 5,640,000, and 55% of them are Chinese. By now the operation is still the best treatment method of PCL and the earlier the better. So the early diagnosis of PCL becomes the key point of the treatment and attracts more and more interest. The aim of this study is to discover new biomarkers of hepatocellular carcinoma (HCC). [METHOD] In this study BALB/c mice were immunized with high metastasis characteristic HCC cells (HCCLM-6). The spleenocytes from the immunized mice were fused with murine myeloma cells (SP2/0) and then the mAb-positive hybridoma cells were screened by indirect ELISA. The mAbs were characterized by immunohistochemical staining in the HCC tissue specimen and indirect immunofluorescent staining in different cell lines. The antigens specifically recognized by the mAbs were identified by Uni-ZAP XR human liver cDNA expression library. [RESULT] Twenty-eight hybridoma cells secreting mAbs were established. Five clones of hybridoma (QAG013, QGA062, QAC081, QAE018 and QFH307) could secret specific mAb against the antigen in HCC. Furthermore, the antigen of the QAG013 was located differently in the tissues and cells. The Uni-ZAP XR human liver expression library was screened by these mAbs. The results indicated that the specific antigen recognized by these mAbs were fibronectin (FN). Combined the results of Western blot and sequencing, it was determined that there were three specific antigen epitopes of FN recognized by these mAbs. Especially the antigen epitope recognized by QGA062 mAb was among 16 amino acids (YTVSLVAIKGNQESPK). [CONCLUSION] Four hybridoma cell lines secreting specific mAb against antigen in HCC were established, and the determination of antigen epitopes of FN would be helpful to the study of new sites in FN associated with HCC.
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