抗Epstein－Bars病毒壳抗原gp125单克隆抗体的研制

Preparation and Application of Anti Epstein-Bars virus gp125 Monoclonal Antibodies

用重组痘苗病毒在１４３细胞上表达的ｇＰ１２５粗提物免疫Ｂａｌｂ／Ｃ小鼠，用杂交瘤技术获得７株稳定分泌抗Ｅｐｓｔｅｉｎ－Ｂａｒｓ（ＥＢ）病毒ｇｐ１２５单克隆抗体的杂交瘤细胞株。对杂交瘤细胞及单克隆抗体进行一系列分析和鉴定，初步建立了用表达产物及特异性单克隆抗体检测ＥＢ病毒ＩｇＡ／ｇｐ１２５抗体的三步ＥＬＩＳＡ法。用此法检测ＩｇＡ／ＶＣＡ阳性的鼻咽癌病人血清及ＩｇＡ／ＶＣＡ阴性的正常人血清，结果完全吻合。

Seven hybridomas secreting McAbs against EBV gp125 were produced by fusion of Sp2/0 myeloma cells with spleen cells from Balb/C mice which were immunized with recombinant antigen. These McAbs showed highly specific activity against gp125with indirect immunofluorescent test. The immunofluorescent titer of McAb in mouse ascitic fluid were 400-1600 and in cell cul ture fluid were 8-32. Six of the McAbs were IgGl subclass and one was Ig2a. Using two McAbs and recombinant antigen, a 3-step ELISA was developed for detecting IgA antibodies to gp125. The IgA antibody to gp125 in 29 sera of patients with NPC and 30 sera of normal individuals was detected by the 3-step ELISA. The positive rate of IgA/gp125 in these two groups was in accordance with that of IgA/VCA by immunoenzymatic test.