分泌抗猪伪狂犬病毒(鄂A株)单克隆抗体杂交瘤细胞株的建立  

Establisment of Hybridoma Cell Strain (EA strain) Producing Monoclonal Antibody to Pseudorabies Virus

作　　者：金梅林[1] 陈焕春[1] 何启盖 方六荣 吴美洲 

机构地区：[1]华中农业大学畜牧兽医学院

出　　处：《中国兽医杂志》1998年第2期3-6,共4页Chinese Journal of Veterinary Medicine

基　　金：湖北省自然科学基金

摘　　要：将本室分离鉴定的猪伪狂犬病毒（Ｐｒｖ）鄂Ａ株纯化后免疫ＢＡＬＢ／Ｃ小鼠，取免疫鼠脾细胞与经８氮鸟嘌呤（８－ＡＧ）驯化的ＳＰ２／０骨髓瘤细胞融合。经间接酶联免疫吸附试验（ＥＬＩＳＡ）筛选，挑选强阳性孔进行亚克隆，获得５株能稳定分泌伪狂犬病毒单克隆抗体（Ｐｒｖ－ＭｃＡｂ）杂交瘤细胞株，分别命名为４１８、４８３、５７６、６０３、７４４。经体外连续传代及反复冻存、复苏，杂交瘤细胞均能稳体地分泌ＭｃＡｂ。培养上清液及小鼠腹水ＥＬＩＳＡ效价分别为２－５～２－７和１０－５～１０－１０。各株分泌的ＭｃＡｂ不与疱疹病毒科中的ＤＰＶ、ＩＬＴＶ、ＩＢＲＶ及ＰＰＶ发生交叉反应。阻断试验结果显示阻断率均大于５０％。其中６０３、７４４分泌的ＭｃＡｂ对用ＰＲＶ致敏的乳胶具有凝集特性。相加ＥＬＩＳＡ测定证实６０３、４８３、７４４特异性作用同一抗原位点或作用相关极为密切的抗原位点，另４种ＭｃＡｂ作用位点有部分相关性。Pseudorabies Virus(A strain) isolated and Identified in our laboratory was purified and used to immunize BALA/C mice .The immunized spleen cells of mice were fused with the SP2/0 cells that were tamed by 8 AG . By detection of ELISA and clonization five hybridoma cell strains of the positive clone secreting Prv McAb were obtained. These five cell strainw were named 418、483、576、603744,which were able to secrete McAbs stablly by series passage cell culturing in vitro and freezing and recovering. The titer of cell culture and ascities tested by ELISA were 2 -5 2 -7 and 10 -5 10 -10 respectively. There was no cross reaction among DPV、ILTV、PPV、IBRV of the three Herpesvidae virus and ppv. The blocking assay showed that their rate of blockig was more than 50%. The secreting McAbs of 603、704 possesse the agglutination activigy to prv sensibilized latex. Additivity ELISA AI results showed that McAbs of 603、483 were specific for the same epitope or very closed related epitopes. The other four McAbs binded somewhat related epitopes.

关 键 词：伪狂犬病毒 单克隆抗体 阻断率 乳胶凝集 ELISA 

分 类 号：S852.65[农业科学—基础兽医学]