抗博尔纳病病毒磷蛋白多克隆抗体的制备及鉴定  

Preparation and identification of polyclonal antibody against phosphoprotein of Borna disease virus

作　　者：马丽华[1,2,3] 黄荣忠[1,2,3] 张亮[1,2,3] 徐晓艳[1,2,3] 曾粒[1,2,3] 刘钊[1,2,3] 邓婧[1,2,3] 李文娟[1,2,3] 谢鹏[1,2,3] 

机构地区：[1]重庆医科大学附属第一医院神经内科,重庆400016 [2]重庆医科大学神经科学研究中心,重庆400016 [3]重庆市神经生物学重点实验室,重庆400016

出　　处：《中国生物制品学杂志》2013年第2期244-247,共4页Chinese Journal of Biologicals

基　　金：国家重点基础研究发展计划(973计划;2009CB918300)

摘　　要：目的制备抗博尔纳病病毒(Borna disease virus,BDV)重组磷蛋白(p24)多克隆抗体,并对其进行鉴定,为BDV血清免疫学检测方法的建立奠定基础。方法用本课题组前期制备的重组BDVp24蛋白经背腿部多点皮下注射免疫新西兰大白兔,共免疫4次,末次免疫后第7天采血,分离血清,采用抗原亲和纯化法纯化抗血清,ELISA法测定抗血清效价;Western blot和免疫荧光鉴定其特异性。结果制备的抗BDV p24多克隆抗体的纯度为98%,ELISA效价达1∶128 000,该抗体与重组p24蛋白和BDV感染OL细胞表达的p24蛋白均能发生特异性反应。结论成功制备了灵敏性和特异性良好的抗BDV p24多克隆抗体,为研发相关的诊断试剂和研究该病毒的感染发病机制奠定了基础。Objective To prepare and identify the polyclonal antibody against recombinant phosphoprotein of Borna disease virus （BDV）, and provide a basis for development of seroimmunological detection method for BDV. Methods New Zealand rabbits were immunized with the recombinant BDV p24 prepared previously by subcutaneous injection in several sites for 4 times, of which the sera were collected on day 7 after the last immunization, purified by affinity chromatography, determined for titer by ELISA, and identified for specificity by Western blot and IFA. Results The prepared polyclonal antibody against BDV p24 showed a purity of 98%, an ELISA titer of 1 ： 128 000, and specific reaction with BDV phosphoprotein expressed either in prokaryotie or in eukaryotic cells. Conclusion The polyelonal antibody against BDV p24, with high sensitivity and specificity, was successfully prepared, which laid a foundation of development of relevant diagnostic kits and study on pathogenic mechanism of BDV.

关 键 词：博尔纳病病毒 磷蛋白类 多克隆抗体 

分 类 号：S852.43[农业科学—基础兽医学] R392.33[农业科学—兽医学]