抗乙型肝炎病毒e抗原和核心抗原双特异性单克隆抗体的建立与应用

用基因工程技术在大肠杆菌中高效表达的乙型肝炎病毒核心抗原(内含高滴度的e抗原)免疫Balb/C小鼠,取免疫小鼠脾细胞与小鼠骨髓瘤细胞(Sp2/0)融合,获得两株分泌高滴度既抗-HBe又抗-HBc的双特异性杂交瘤细胞系。细胞培养上清液中抗体滴度为100～1000以上;免疫腹水中的抗体滴度为8万至10万以上,均属IgG2a亚类。细胞在实验室连续传代二年多,仍保持高效分泌抗体能力。此单克隆抗体与HBeAg或HBcAg的结合可被抗-HBc或抗-HBc阳性血清所抑制,竞争抑制率在85.9％～96.8％之间。用此单克隆抗体与HBe的β型单克隆抗体和抗HBc的α型单克隆抗体配对,可组装成检测HBeAg/抗-HBe和抗-HBc的诊断试剂,具有重要的应用价值。

ESTABLISHMENT AND APPLICATION OF TWO HYBRIDOMAS PRODUCING BISPECIFIC MONOCLONAL ANTIBODIES TO HBeAg AND HBcAg

Hepatitis B viral c antigen ( contain HBeAg ) expressed in E.coli was used to immune Balb/C mice. The immunized mouse spleen cells were fused with mouse myeloma cells ( Sp2/0 ) by PEG. Two hybridomas ( lG8, 1D3) that secrete antibodies to both HBeAg and HBeAg were established. The antibodies ELISA titer were 105 in mouse ascitic fluids. Both of them were IgG2a subclass. These McAbs only react with HBeAg and HBc Ag but do not react with normal human serum and extracts from E.coli. The reaction between bispecific McAbs and antigens can be competed with anti-HBe and anti-HBc positive sera respectively. These McAbs have high specificity, are highly stable and have good prospects of application either to develop diagnostic reagents or for research on the relationship between HBeAg and HBeAg. The bispecific McAbs combined with McAb against HBeAg can be used to develop a diagnostic reagent for testing HBeAg and anti-HBe. If combined with McAb to HBeAg, it can be used to develop a diagnostic reagent for testing anti-HBc antibody.So far, it has been used to coat polyvinyl plates or beads, or labelled with horseradish peroxidase ( HRP ) or125I and used in combination with monoclonal antibody to HBeAg ( C4 etc. established in our lab. ) for testing HBeAg and anti-HBe in China.