摘要
目的建立能稳定分泌抗丙型肝炎病毒(HCV)NS5重组蛋白单克隆抗体(McAb),并对其生物学活性进行实验室鉴定。方法用重组HCVNS5区蛋白为抗原免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0进行融合,经有限稀释克隆3次后制备分泌McAb的杂交瘤细胞株,并用酶免疫(EIA)法对其分泌抗体进行初步鉴定。结果融合后的阳性克隆中筛选出6株能稳定分泌McAb的杂交瘤细胞株,命名为2C8,2D2,1D6,2G2,1F5和1F10。这6株McAb与NS5重组抗原均有良好的反应性,杂交瘤培养上清的EIA抗体滴度为1∶400~1∶1600,其中1株诱生的同系小鼠腹水滴度为1∶80000。这6株McAb均为IgG1亚型。结论该McAb的制备,可为HCV感染者血清和肝组织中抗原检测方法的建立和生物工程药物的研制创造条件。
ObjectiveThe goal is to establish hybridoma cell line which can secrete monoclonal antibodies against HCV recombinant NS 5 protein and to determine biological activity of the monoclonal antibodies. MethodsThe BALB/c mice were immunized through intraperitoneal injection of the NS 5 region protein of hepatitis C virus. After three times injection, the splenocytide positive for the NS 5 region antigen was fused with SP 2/0 mouse myeloma cells. To prepare secretable McAb byhridoma cell lines, three times subcloning method, limited dilution method, were used. The antibody strains were analyzed by EIA. ResultsSix hybridoma cell lines of positive for HCV NS 5 protein were selected designed 2C 8,2D 2, 1D 6, 2G 2,1F 5 and 1F 10 from cloning strains. The titer of EIA of the culture supernatant were 1∶400~1∶ 1 600 . The titer of EIA in one animal of ascites was 1∶ 80 000 . All the monoclonal 1 antibody strain was IgG 1 subtype. ConclusionThe McAb can be used in detection of HCV antigen in sera and hepatocyte and used in study of new bioengineering medicine.
出处
《河北医科大学学报》
CAS
1999年第4期220-222,共3页
Journal of Hebei Medical University
基金
河北省科委和河北省卫生厅联合资助
关键词
单克隆抗体
分离
提纯
HCV
NS5区
重组蛋白
MeSH antibodies
monoclonal/isol
hepatitis virus/isol
HCV NS 5 recombinant protein/methods
mice