摘要
为制备猪伪狂犬病毒(Pseudorabies virus,PRV)变异毒株的单抗,以纯化灭活的PRV JS-2012毒株免疫BALB/c雌性小鼠。经间接免疫荧光法(indirect immunofluorescence assay,IFA)筛选,获得2株稳定分泌PRV单抗的杂交瘤细胞:4D8、4F10。2株单抗经亚型鉴定均为Ig G1亚类,轻链为κ链,腹水IFA效价均达1:51 200;IFA与IHC结果显示,二者均能与PRV毒株发生特异性反应;Western blot结果表明,4D8与PRV反应条带大小约为60 k Da,而4F10不与PRV发生反应,推测其可能针对PRV的构象性抗原表位。该单抗的制备对PRV的抗原变异及诊断研究均有重要意义。
In order to generate the monoclonal antibodies(MAbs) against a variant of Porcine pseudorabies virus(PRV), BALB/c mice were immunized with purified and inactivated PRV JS-2012 strain. Two hybridomas secreting PRV specific MAbs were obtained and designated as 4D8 and 4F10, respectively. Both MAbs belonged to IgG1 subgroup and had κ light chains. The ascetic fluids containing either 4D8 or 4F10 showed indirect immunofluorescence assay(IFA) titers at 1:51200. Both MAbs reacted specifically with PRV in IFA and immunohistochemical(IHC). In Western blot, MAb 4D8 reacted with 60 k Da protein of PRV while MAb 4F10 showed no obvious reactions with PRV protein, suggesting that MAb 4F10 might react with a conformational epitope. The availability of PRV MAbs was important for further research of antigenic variation and diagnosis.
出处
《中国动物传染病学报》
CAS
北大核心
2015年第3期7-11,共5页
Chinese Journal of Animal Infectious Diseases
基金
上海市自然科学基金项目(14ZR1448900)
中央级公益性科研院所基本科研业务费专项资金项目(2014JB02)
关键词
伪狂犬病毒
单克隆抗体
间接免疫荧光检测
免疫组化鉴定
Pseudorabies virus
monoclonal antibody
indirect immunofluorescence assay (IFA)
immunohistochemical (IHC)