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目的制备丙型肝炎病毒(HCV)单克隆抗体(Mc Ab),为HCV抗原诊断试剂的研发奠定基础。方法用纯化的含NS4抗原位点及NS5抗原位点的基因工程表达的融合蛋白(HCAg)免疫Balb/c小鼠,取脾细胞及骨髓瘤细胞按常规方法融合、筛选、克隆及腹腔注射,制备Mc Ab,用ELISA进行特异性鉴定,使用亲和层析法纯化。结果筛选出4株能稳定分泌特异性抗-HCV的Mc Ab杂交瘤细胞株,经多次复苏、传代仍能稳定分泌抗体,特异性强、效价高。间接免疫荧光显示,4株杂交瘤细胞株分泌的抗体对HCAg具有特异亲和性。结论获得了4株抗-HCV的Mc Ab,为今后研发HCV抗原诊断试剂打下了基础。
Objective To prepare a monoclonal antibody against Hepatitis C virus (McAb), which lays a foundation for the development of HCV antigen diagnostic reagents. Methods Balb / c mice were immunized with the purified fusion protein (HCAg) expressed by NS4 antigen and NS5 antigen. The spleen cells and myeloma cells were fused, screened, cloned and injected intraperitoneally. Mc Abs were prepared, specifically identified by ELISA, and purified using affinity chromatography. Results Four strains of McAbs that could stably secrete specific anti-HCV were screened out. After repeated resuscitation, the McAbs still secreted steadily with high specificity and high potency. Indirect immunofluorescence showed that the antibodies secreted by the four hybridoma cell lines had a specific affinity for HCAg. Conclusions Four McAbs against HCV were obtained, which laid the foundation for the future development of HCV antigen diagnostic reagents.