【摘要】 为制备抗犬I型腺病毒单克隆抗体,将犬I型腺病毒(CAV-I)细胞培养液饱和硫酸铵沉淀,差速离心浓缩,氯化铯密度梯度离心的方法纯化后免疫BALB/c小鼠,三免后效价过1:10000即可取脾细胞与SP2/0细胞在聚乙二醇(PEG)作用下融合,通过间接ELISA方法筛选阳性杂交瘤细胞株,有限稀释法亚克隆,制备单克隆抗体,并对制备完成的单克隆抗体进行生物学特性鉴定。获得2株能稳定分泌抗CAV-I的单克隆抗体杂交瘤细胞,命名为C8、E9,经鉴定其亚型分别为IgG1和IgG2a。间接ELISA检测效价,2株单抗细胞上清液效价为1:3200～1:6400,腹水效价为1:25600～1:51200。该单克隆抗体与CDV、FPV、FCV病毒均无交叉反应。实验成功制备了抗CAV-I单克隆抗体,为进一步建立相关诊断方法奠定了基础。更多还原

【Abstract】 To prepare and identify the monoclonal antibody against canine adenovirus typeⅠ.Cell culture fluid of CAV-I were precipitationed by saturated ammonium sulfate,concentrated differential centrifugation,caesium chloride density gradient centrifugation,BALB/c mice were immunized conventionally with the purified CAV-I.On the third day after the final immunization,spleen cells of mice fused with myeloma cells SP2/0 were filtered by culturing selectively,detecting special antibody,ELISA test,and coloning.Two hybridoma cell lines,obtained by culturing in vitro and anabiosis after freezing in a long period which were named C8,E9,could stably secrete McAbs.The subclasses of the two McAbs were IgG2a and IgG1 respectively.Indirect ELISA titer detected,the titers of cell cultures ranged from 1:3200 to 1:6400 and the titers of ascites fluids ranged from 1:25600 to 1:51200.These McAbs were specific to CAV-I,but not reacted with CDV,FPV,FCV.McAbs against CAV-I has been successfully prepared.It is also a basic for establishing a related diagnoses method.更多还原