摘要
目的建立能稳定分泌抗乙型肝炎病毒核糖核酸酶H1及H2(HBVRNaseH1及H2)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HBVRNaseH1及H2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4株能稳定分泌McAb的杂交瘤细胞株,命名为H1C6、D3;H2E2、F4。这4株McAb与HBVRNaseH1及H2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∶100～1∶200,其中2株诱生的同系小鼠腹水滴度为1∶800,1∶1000。这4株McAb均为IgG1亚型。结论该McAb的制备,可为HBV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。
Objective To establish hybridoma cell line which can secret e monoclonal antibodies against HBV recombinant RNaseH_1) and H_2 protein,an d to determine biological activity of the monoclonal antibodies. Methods The BALB/c mice were immuni zed through intraperitoneal injection of the RNase H_1 and H_2 protein of hepatitis B virus. After thr ee times injection, the splenocytide positive for the RNase H_1 and H_2 antigen was fused with SP 2/0 mouse myeloma cells. To prepare secretable McAb by hridoma cell lines, three times subcloning and, limi ted dilution method were used. The antibody strains were analyzed by EIA. Results Four hybridoma cell lines of HBV-RNaseH H_1 C6, D3;H_2 E2, F_4 were selected from cloning strains. The titer of EIA of the culture supernatant was 1:100～1:200. The titer of EIA of ascites in two animal was 1∶800, 1∶1000. All the monoclonal antibo dy strain was IgG1 subtype. Conclusion The McAb can be used in detection of HBV antigen in sera and hepatocyte and used in study of new bioengineering medicine.
出处
《胃肠病学和肝病学杂志》
CAS
2005年第3期223-226,共4页
Chinese Journal of Gastroenterology and Hepatology
