鸭肠炎病毒VP5蛋白单克隆抗体的制备及鉴定  

Development and identification of the monoclonal antibodies against duck enteritis virus VP5 protein

作　　者：李慧昕[1,2] 刘胜旺[2] 韩宗玺[2] 邵昱昊[2] 刘晓丽[2] 华育平[1] 刘娣[1] 孔宪刚[2] 

机构地区：[1]东北林业大学博士后流动站黑龙江省农业科学院博士后工作站,黑龙江哈尔滨150086 [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禽传染病研究室,黑龙江哈尔滨150001

出　　处：《中国兽医杂志》2013年第1期9-12,共4页Chinese Journal of Veterinary Medicine

基　　金：黑龙江省自然科学基金(QC08C12);中国博士后科学基金(2008440920);黑龙江省博士后资助项目(LBH-Z08022)

摘　　要：应用截短表达的鸭肠炎病毒(DEV)VP5-C蛋白免疫BALB/c小鼠,通过细胞融合及筛选,获得4株稳定分泌抗VP5蛋白的杂交瘤细胞株,命名为1B5、3A1、4F9、6F6。抗体亚类鉴定表明,4株单克隆抗体均为IgG、k链。间接免疫荧光检测结果表明,4株单克隆抗体均能够与DEV感染CEF发生特异性反应,而不与CEF对照发生反应。Western blot检测,4株单克隆抗体均能够识别大小约为150kDa的蛋白,说明4株单抗隆抗体是特异性识别VP5蛋白的抗体。DEV VP5蛋白单克隆抗体的制备为进一步鉴定VP5蛋白B细胞抗原表位及VP5蛋白功能研究提供了有效工具。The purified protein of truncated VP5-C was used to immunize the BALB/c mice.Four stable hybridoma cell strains secreting antibody against VP5 were screened and designated as 1B5、3A1、4F9 and 6F6,respectively.Four hybridoma cell strains were detected as IgG,κ chain by the subtype identification assay.The indirect immunofluorescent assay revealed that four McAbs could detect the DEV-infected CEF but not CEF controls.Western blot detection showed that the four McAbs could recognize the VP5 protein with a molecular weight 150 KDa specifically.The development of McAbs against VP5 protein provided a useful tool for the study on the B cell epitopes and the function of VP5 during the virus life cycle.

关 键 词：鸭肠炎病毒 VP5蛋白 单克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]