论文部分内容阅读
目的制备抗轮状病毒(RV)LLR株G10血清型特异性单克隆抗体,并鉴定其特性。方法以纯化的RVLLR株(血清型为G10)为抗原免疫BALB/c小鼠,应用杂交瘤细胞技术筛选分泌G10血清型特异性单抗的杂交瘤细胞株。采用层析法纯化单抗,常规免疫学方法鉴定单抗的特性。结果经筛选获得5G5、1H1和7F6共3株可稳定分泌高效价且具有G10血清型特异性单抗的杂交瘤细胞株。3株细胞分泌的单抗分别为IgG2a/κ、IgG2a/κ和IgG1/κ型;腹水效价均可达1.024×106;分别识别两个抗原位点;相对亲和力依次为1H1﹥7F6﹥5G5;5G5和1H1(细胞上清)均与G1～G4和G6血清型RV抗原无交叉反应;3株杂交瘤细胞分泌的单抗均识别相对分子质量约为34000的VP7蛋白,且均有不同程度的中和活性。结论已成功制备了抗RVLLR株G10血清型特异性单抗,该单抗可用于G10血清型RV的分型检测或LLR毒株的鉴别。
Objective To prepare serotype specific monoclonal antibodies against rotavirus (RV) LLR strain G10 and identify its characteristics. Methods BALB / c mice were immunized with the purified RVLLR strain (G10 serotype) as an antigen and the hybridoma cell lines secreting G10 serotype specific monoclonal antibodies were screened by hybridoma technique. Monoclonal antibodies were purified by chromatography and identified by routine immunological methods. Results A total of 3 strains of 5G5, 1H1 and 7F6 were screened, which could stably secrete high titer hybridomas with G10 serotype specific McAbs. The McAb secreted by the three strains were IgG2a / κ, IgG2a / κ and IgG1 / κ, respectively. The ascites titer reached 1.024 × 106. The two antigenic sites were identified respectively. The relative affinities were 1H1> 7F6> 5G5 and 1H1 (cell supernatant) did not cross-react with RV antigen of G1-G4 and G6 serotypes. The monoclonal antibodies secreted by the 3 hybridoma cells both identified VP7 protein with a relative molecular mass of about 34000 and all had different degrees of Neutralize activity. Conclusion G10 serotype specific monoclonal antibody against RVLLR strain was successfully prepared and could be used to detect the genotype of G10 serotype RV or identify LLR strains.