【作者】 温贵兰； 李永明；

【Author】 WEN Gui-lan, LI Yong-ming(College of Animal Science, Guizhou University, Guiyang 550025, Guizhou, China)

【机构】 贵州大学动物科学学院；

【摘要】 以蔗糖反透析浓缩,Sephadex G-200层析纯化F₄₈E₈和Lasota新城疫病毒（NDV）。取纯化的病毒液免疫接种Balb/c小鼠,将小鼠脾细胞与NS-1骨髓瘤细胞融合,经次黄嘌呤、氨基喋呤和胸苷（HAT）培养基选择。间接ELISA检测,有限稀释法克隆,筛选出3株杂交瘤细胞,依次为1D₄、1D₅、16G₁₂。3株杂交瘤细胞的染色体数在80～100之间,并能稳定传代。其产生的单克隆抗体重链属于IgG₁,轻链属于κ链;在ELISA检测中仅与NDV发生特异性反应与其他常见禽类病毒抗原不出现交叉反应。应用这些单克隆抗体对本实验室保存的NDV分离株进行了分群研究。更多还原

【Abstract】 The chicken embryo allantoic fluid infected with NDV was concentrated by reverse dialysis of sucrose. The concentrated virus was purified by chromatography column of Sephadex G-200. Three strains of hybridoma cells secreting monoclonal antibody（McAb） were obtained by the fusion of NS-1 myeloma cell line and spleen cell from mice immunized with F₄₈ E₈ and Lasota, cultivated in HAT medium, screened with ELISA and cloned by the limited dilution. The three strains of hybridoma cells were designated as 1D₄, 1D₅, 16G₁₂ respectively. The number of chromosome ranged is from 80 to 100. The McAbs were secreted from the three strains of hybridoma cells after being cultivated two months. The H-chain of three McAbs belonged to IgG₁ and L-chain belonged to κ type. They were specific for NDV and nocross reactions were observed with other avian virus antigens. The NDV isolates kept in this lab were divided into three groups with three McAbs.更多还原