新城疫病毒F48E9株重组NP蛋白单克隆抗体制备及鉴定  

Preparation and identification of monoclonal antibody against recombinant NP protein of NDV F_(48)E_9

作　　者：刘文丽[1,2,3] 刘怀然[2] 刘培欣[2] 张馨心[1] 孔宪刚[2] 陈维多[1] 

机构地区：[1]东北农业大学生命科学学院,哈尔滨150030 [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,哈尔滨150001 [3]黑龙江中医药大学,哈尔滨150040

出　　处：《东北农业大学学报》2011年第9期131-135,147,共6页Journal of Northeast Agricultural University

基　　金：国家科技支撑计划(2006BAD06A03,2006BAD06A08)

摘　　要：制备新城疫病毒(NDV)NP蛋白单抗,以期为NDV抗原表位研究及检测方法建立方面奠定基础。利用NDV F48E9株pET32a-NP重组蛋白免疫BALB/c小鼠,采用杂交瘤细胞技术,间接ELISA筛选,获得了2株稳定分泌抗NP重组蛋白单克隆抗体的杂交瘤细胞株,分别命名为1G3、4B12。经间接ELISA测定,腹水抗体效价分别为1 2.56×105、1 5.12×105。亚类鉴定结果表明这两株单抗均为IgG1。Western blot分析结果显示,1G3、4B12均能特异性识别重组NP蛋白。与NDV感染细胞经间接免疫荧光试验检测均呈黄绿色荧光。经相加ELISA测定表明两株单抗识别的抗原表位不同。Newcastle disease virus（NDV）was prepared in order to lay the foundation for NDV antigen epitope research and establishment of detection methods.The resnlt showed that BALB/c mice were immunized with pET32a-NP recombinant protein of NDV F48E9,two hybridoma cell lines steadily secreting McAb against NP protein of NDV were obtained by hybridomatechnology and indirect ELISA,and the cell lines were named 1G3 and 4B12.Their titers of ascites were 1 2.56×105 and 1 5.12 ×105,respectively,by indirect ELISA.The isotypes of McAbs were all IgG1.They were highly specific to recombinant NP protein by Western blot.It showed yellow and green fluorescence that cells infected with NDV by Indirect immunofluorescence assay（IFA）.Additive ELISA showed that the two McAbs could recognize different epitopes.

关 键 词：新城疫病毒 F48E9 NP蛋白 单克隆抗体 

分 类 号：S852.659.5[农业科学—基础兽医学]