论文部分内容阅读
制备高特异性的抗鲤春血症病毒单克隆抗体,并对其免疫学特性进行鉴定。采用差速离心法提纯的鲤春血症病毒作为免疫原,免疫Balb/c小鼠,4次免疫后,利用杂交瘤细胞技术进行细胞融合,经过多次亚克隆筛选出高敏感的特异性抗鲤春血症病毒单克隆抗体杂交瘤细胞株,体内诱生腹水制备抗鲤春血症病毒单克隆抗体并对其免疫学特性进行鉴定。融合后筛出两株杂交瘤细胞株9C11和1C12,亚型均为IgG2b型,κ链;间接ELISA测定腹水效价为104,与其他病毒株和细胞株不发生反应;western-blot结果显示单克隆抗体对应的抗原位点在相对分子质量(Mr)230 000的条带处;免疫氧化酶染色结果显示单克隆抗体能够特异地识别感染细胞内的鲤春血症病毒。本实验制备的抗鲤春血症病毒单克隆抗体特异性好,为鲤春血症病毒免疫学快速检测方法的建立奠定了基础。
Preparation of high specificity of anti-carp virus hemagglutinin monoclonal antibody, and its immunological properties were identified. Balb / c mice were immunized with common carp haemocytoma virus purified by differential centrifugation. After four immunizations, hybridoma cells were used for cell fusion. Subsequent clones were screened for high sensitivity and specificity Common carp virus hemagglutinin monoclonal antibody hybridoma cell line, in vivo induced ascites anti-common carnitine virus monoclonal antibody and immunological characteristics were identified. After hybridization, two hybridoma cell lines, 9C11 and 1C12, were screened, and the subtypes were all IgG2b type and κ chain. The indirect ELISA assay showed that ascites titer was 104 and did not react with other virus strains and cell lines. Western- The corresponding antigenic site of the cloned antibody was at a band of 230 000 with molecular weight (Mr). Immunooxidase staining showed that the monoclonal antibody could specifically recognize the common carp virus in the infected cells. The specificity of the monoclonal antibody against the anti-carp virus of the carp virus prepared in this experiment lays a foundation for the establishment of a rapid immunological detection method for the common carp virus.