摘要
应用杂交瘤技术,以大肠杆菌基因工程表达的丁型肝炎病毒抗原(HDAg)免疫Balb/C小鼠,首次建立了6株分泌抗丁型肝炎病毒(HDV)单克隆抗体细胞株。它们的培养上清和腹水的抗体滴度分别是50～1000和1000～40000。Ig亚类4株为IgG1,1株IgG2a,1株IgG。这些单克隆抗体能和HDAg结合,而不与大肠杆菌裂解液,大肠杆菌表达的HBcAg、HBeAg,以及HBsAg阳性血清和正常人血清发生反应。它们与HDAg的结合能被抗HD阳性血清所阻断,说明这种结合是特异的。这些McAb的特异性和亲和性与多克隆抗体相同。以McAb做包被抗体和抗HDV多克隆酶标抗体配对制成EIA试剂,检测血清标本中的HDAg和抗-HD,与PcAbEIA的符合率分别为94.44％和98.68％。以PcAb包被,配以酶标记的McAb;或以McAb包被,配以酶标记的PcAb均可检测大肠杆菌表达的HDAg,而且效果都优于PcAb-EIA。用这些McAb对国外基因工程表达的HDAg进行的抗原位点分析首次证明,在这些HDAg上至少有3个不同的抗原位点。通过对基因工程表达的HDAg和我国人血清中HDAg的抗原位点的分析,认为两者至少在空间构型上有不同。
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.
出处
《病毒学报》
CAS
CSCD
北大核心
1992年第1期7-12,共6页
Chinese Journal of Virology
基金
国家"863"高技术生物技术领域资助项目
关键词
单克隆抗体
丁肝病毒
Hepatitis D virus Monoclonal antibodies against HDV
