Production of monoclonal antibody specific to surface antigen on hepatocellular carcinoma cell lines

Abstract

Liver cancer ranks fifth in frequency in the world and this disease tends to increase every year. HCC is a type of liver cancer which is the most severe. When there are signs and symptoms, it means poor prognosis. Patients always die within a few months after detecting the nodules. In general, HCC is diagnosed as high level of AFP. However, because of its low specificity, expensive radiologic imaging is included as confirmatory test. Diagnosis of HCC still relies on liver biopsy. The objective of this study was to produce monoclonal antibody specific to surface molecule on HCC cells in order to apply as another tumor specific marker for diagnosis of HCC. BALB/c mouse was immunized i.p. weekly with cold inactivated HepG2 cells. The rising of antibody was determined by indirect immunofluorescent technique using HepG2 cells as source of antigens. Immunized mouse’s spleen cells were fused with myeloma cells (X63Ag8.653) by standard hybridoma technology and limiting dilution. Cell culture supernatant from hybrid clones were investigated for anti-HepG2 cells antibody by indirect immunofluorescent technique and analyzed by flow cytometry. One hundred and ten hybrid cells (22.9%) were obtained with 38 clones (7.9%) produced anti-HepG2 antibodies as investigated by indirect immunofluorescent technique. All tested clones could react with HepG2 both in suspension and adherent condition suggesting that they could be applied to use in immunohistochemistry technique as objected. The obtained monoclonal antibodies will be further on investigated for their specifity to tumor antigen or tumor associated antigen.