抗丁型肝炎病毒单克隆抗体的制备及其生物学特性

PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO HEPATITIS DELTA VIRUS

Hepatitis delta virus(HDV) is a defective single-strand RNA virus that requires hepatitis B virus for its replication and occurs in patient with acute and chronic HBV infection.We have established six hybri doma cell lines by fusion immunized mouse spleen cells with SP2/0 myeloma cell. All of them secret monoclonal antibody against HDAg that can be used to develop diagnostic reagents and analyse the anti genie domains of HDV.The titer of McAb in culture fluid and ascites fluids were 1:50 to 1: 1,000 and 1:1,000 to 1 : 40,000 respectively. Four of them were IgG1, one was IgG2a and another was IgG mouse immu noglobulin. McAbs can react with HDAg in serum ( S-HDAg ) or expressed in E.coli(E-HDAg)but did not react with extract of E.coli,HB cAg and HBsAg expressed in E.coli, HBsAg positive serum and normal human serum.The reaction of McAbs with HDAg can be blocked by anti-HD positive human serum. This result shows that these McAbs were specific for HDAg. McAbs were used as a coating antibody for the detection of HDAg and Anti-HD in sera and compared with the results using polyclonal antibody. The coincident rate were 94.44% and 98.68% respectively.The result of detecting E-HDAg by McAb-EIA was much better than PcAb-EIA. We have found at least three antigenic determinatants on E-HDAg by McAbs to HDAg.The results also showed that the conformational structure of E-HDAg may be different from that of S-HDAg by analysis of antigenic domains with McAbs.