【摘要】 在MDBK细胞上增殖牛病毒性腹泻病毒(BVDV)HN 1 株,用蔗糖密度梯度离心法纯化后免疫BALB/c小鼠,应用淋巴细胞杂交瘤技术,取脾细胞并与NS0 骨髓瘤细胞融合,经间接ELISA筛选,3次有限稀释法克隆,得到4株能稳定分泌抗BVDV单克隆抗体(McAb)的杂交瘤细胞株:3A8、3C7、3C11和3E9。经鉴定,3A8、3C7、3E9株为IgG1亚类,3C11株为IgG2a亚类;杂交瘤细胞的平均染色体数目为99 条。3A8、3C7、3E9、3C11 与BCV、BRV均无交叉反应,但3A8、3C7、3E9与HCV、BDV存在交叉反应,而3C11与HCV、BDV无交叉反应,3C11显示出较好的特异性;杂交瘤细胞培养上清液及小鼠腹水McAb的ELISA效价分别为1∶1 000 和1∶200 000,该杂交瘤细胞连续培养20代后仍能稳定分泌抗体。这些McAb的获得为BVDV的研究及快速诊断方法的建立奠定了基础。更多还原

【Abstract】 MDBK cells were infected with BVDV HN-1.The virus was centrifugated and purified. BALB/c mice were immunized with the purified BVDV. Using lymphocyte hybridoma technique,mouse spleen cells were fused with myelomas cells NS0.Four hybridoma cell strains which can excrete stably monoclonal antibody were obtained by indirect ELISA and tri-time limiting dilution.The four hybridoma cell strains are named 3A₈,3C₇,3C₁₁ and 3E₉ respectively. By evaluating,three strains belong to IgG1 subset, and the other one is IgG2a subset.The average chromatosome number of hybridoma cells are 99 strips. The monoclonal antibodies secreted by 3A₈,3C₇,3C₁₁ and 3E₉ strains have no cross reactions with BCV and BRV. The monoclonal antibody secreted by 3A₈,3C₇,3E₉ strains have cross reactions with HCV,BDV,but the monoclonal antibody secreted by 3C₁₁ strain has no cross reactions with HCV and BDV. The specificity of monoclonal antibody secreted by 3C₁₁ strain is better. ELISA titers of monoclonal antibodys which are existent in superior schedule of culture of hybridoma cells and ascites of mice are 1∶1000 and （1∶200000） respectively. After being continuously cultivated to 20 generations, the hybridoma cell strains could still excrete antibody stablely.The monoclonal antibody was very important to the investigation of BVDV and the creating pykno-diagnosis method.更多还原