抗犬Ⅰ型腺病毒单克隆抗体的制备及生物学特性鉴定  

Preparation of Monoclonal Antibodies against Canine Adenovirus TypeⅠ and Their Identification of Biological Characters

作　　者：李晓叶[1,2] 刘颖[1,2] 蓝田丰[1,2] 艾纯旭[1,2] 陈健[2] 袁宝[2] 刘殿峰[2] 任文陟[2] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]吉林大学实验动物中心,长春130062

出　　处：《中国农学通报》2011年第11期31-34,共4页Chinese Agricultural Science Bulletin

基　　金：吉林省科技平台建设项目"吉林省实验动物质量检测中心平台建设"(20071138)

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。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.

关 键 词：犬I型腺病毒 杂交瘤 单克隆抗体 酶联免疫法 鉴定 

分 类 号：Q511[生物学—生物化学]