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抗Asia1型口蹄疫病毒单克隆抗体的制备及生物学特性鉴定
引用本文:李万,周国辉,杨增岐,涂亚斌,于力.抗Asia1型口蹄疫病毒单克隆抗体的制备及生物学特性鉴定[J].中国预防兽医学报,2007,29(6):448-452.
作者姓名:李万  周国辉  杨增岐  涂亚斌  于力
作者单位:1. 西北农林科技大学,动物科技学院,陕西,杨凌,712100;中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室/大动物病研究室,黑龙江,哈尔滨,150001
2. 中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室/大动物病研究室,黑龙江,哈尔滨,150001
3. 西北农林科技大学,动物科技学院,陕西,杨凌,712100
摘    要:用纯化的Asia1型口蹄疫病毒免疫BALB/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行融合,经间接ELISA和间接免疫荧光(IFA)筛选,有限稀释法克隆,获得了2株稳定分泌单克隆抗体的杂交瘤细胞株,分别命名为3H6、5G3,其细胞培养上清效价分别为1:64和1:128,小鼠腹水效价分别为1×10~(-4)和8×10~(-3);ELISA和IFA结果显示,2株单抗仅与Asial型口蹄疫病毒反应,不与O型口蹄疫病毒反应,表明它们均为抗Asial型口蹄疫病毒的型特异性单克隆抗体。westem blot结果显示,2株单克隆抗体均不与全病毒抗原反应,表明它们所针对的抗原表位均为构象表位。相加ELISA试验表明,两株单抗识别不同的抗原表位。经硫氰酸盐洗脱法测定,3H6和5G3的相对亲和力指数分别为1.0 mol/L和1.5 mol/L。这2株单抗的获得为建立口蹄疫病毒检测方法提供了强有力的工具。

关 键 词:口蹄疫病毒  单克隆抗体
文章编号:1008-0589(2007)06-0448-05
修稿时间:2007-01-12

Production and characterization of monoclonal antibodies against Asia1 foot-and-mouth disease virus
LI Wan,ZHOU Guo-hui,YANG Zeng-qi,TU Ya-bin,YU Li.Production and characterization of monoclonal antibodies against Asia1 foot-and-mouth disease virus[J].Chinese Journal of Preventive Veterinary Medicine,2007,29(6):448-452.
Authors:LI Wan  ZHOU Guo-hui  YANG Zeng-qi  TU Ya-bin  YU Li
Institution:1. College of Animal Science and Technology, Northwest Sci-Tech University of Agriculture and Forestry, Yangling 712100, China; 2. Division of Livestock Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China
Abstract:Monoclonal antibodies(McAbs)3H6 and 5G3 against FMDV(Foot-and mouth disease virus)serotype Asial were produced by fusing SP2/0 myeloma cells with spleen cells of BALB/c mice immunized with purified viruses.Both McAbs reacted with FMDV serotype Asial but not with FMDV serotype O,as determined by Enzyme-linked immunsorbent assay(ELISA) and indirect immunofluorescence assay(IFA).Western blot analysis showed that both McAbs recognized conformational epitopes but not whole viral proteins.The relative affinity index(RAI)of the McAbs were 1.0 mol/L and 1.5 mol/L,respectively,as indicated by thiocyanate elution measurement.The result of additive ELISA showed that the McAbs could recognize different antigen epitopes.The McAbs are useful tools for developing diagnostic techniques of foot-and-mouth disease virus serotype Asial.
Keywords:foot-and-mouth disease virus  monoclonal antibodies
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