论文部分内容阅读
目的制备抗病毒性出血性败血症病毒(VHSV)的单克隆抗体(mAb)。方法以差速离心法提纯VHSV作为免疫原,免疫BALB/c小鼠。应用杂交瘤细胞技术进行细胞融合,间接ELISA进行筛选检测,且对mAb进行特性分析。结果制备出1株抗VHSV mAb,命名为4A5。对其进行特性分析结果显示该抗体腹水效价104以上;属于IgG3型,κ链;仅与VHSV本身反应,与其他病毒和细胞均不发生交叉反应;对应的抗原位点在VHSV蛋白的相对分子质量(Mr)70 000的条带处,该蛋白带是VHSV的G蛋白。结论制备出的特异性好的抗VHSV mAb,为该病毒的快速诊断及流行病学监测提供了检测试剂。
Objective To prepare a monoclonal antibody (mAb) against the viral hemorrhagic septicemia virus (VHSV). Methods VHSV was purified by differential centrifugation and immunized BALB / c mice. Hybridization of cells using hybridoma cell technology, indirect ELISA screening test, and mAb characterization. Results An anti-VHSV mAb was prepared and named 4A5. The results of their characterization showed that the antibody had an ascitic titer of 104 or more, belongs to IgG3 type and κ chain, only reacts with VHSV itself, does not cross-react with other viruses and cells, and the corresponding antigenic site is at the VHSV protein relative molecule At a band of 70,000 Mr, the protein band is a VHSV G protein. Conclusion The prepared anti-VHSV mAb has provided the detection reagent for the rapid diagnosis and epidemiological surveillance of the virus.