畜牧与兽医2018，Vol.50Issue(1)：109-112,4.
猪伪狂犬病毒gE蛋白单克隆抗体制备与鉴定
Preparation and identification of monoclonal antibodies against gE protein of pseudorabies virus
摘要
Abstract
In order to obtain the pseudorabies virus (PRV) gE protein monoclonal antibody,the recombinant gE protein by prokaryotic expression was immunized into female BALB/c mice aged 6 weeks,the spleen cells and SP2/0 were fused,and the positive hybridoma cells were screened by indirect ELISA.The results showed that two hybridoma cell lines secreting anti-PRV gE protein were obtained,named E3B8 and E5C11,respectively.The antibody titer in the supernatants of the two hybridoma cells was 1 ∶ 6.4× 103,while the titers in ascites reached 1 ∶ 3.28×106 and 1 ∶ 6.55× 106,respectively.The chromosome numbers of the two hybridoma cells were 105 and 108.The heavy chain subtypes of E3B8 and ESC11 were IgG1 and IgG2b,respectively,with a κ light chain.Western blot analysis showed that the two monoclonal antibodies could specifically react with the recombinant gE protein of PRV.Indirect immunofluorescence assay (IFA) detection showed that the two monoclonal antibodies could specifically react with the infected BHK-21 cells with PRV.And the two monoclonal antibodies did not cross-react with other viruses.It suggests that the two monoclonal antibodies established in this study had high titer and specificity,which laid the foundation for the research on the structure and function of gE protein and the development of PRV immunological diagnostic kit.关键词
伪狂犬病毒/gE蛋白/单克隆抗体/制备/鉴定Key words
pseudorabies virus/gE protein/monoclonal antibodies/preparation/identification分类
农业科技引用本文复制引用
雷有玲,李国泰,鲍玉林,刘秀清,张文..猪伪狂犬病毒gE蛋白单克隆抗体制备与鉴定[J].畜牧与兽医,2018,50(1):109-112,4.基金项目
基余项目:青海省农技推广补助项目(青农发[2015] 293号) （青农发[2015] 293号）
