传染性法氏囊病病毒抗原表位分析——单克隆抗体的制备与鉴定  

Analysis of the antigenic epitopes of infectious bursal disease virus-establishment and identification of monoclonal antibodies to infectious bursal disease virus

作　　者：王军 李银[2] 范红结[3] 周宗安[4] 施正良[4] 王永山[4] 

机构地区：[1]山东省淄博市兽医站,山东淄博255026 [2]江苏省农业科学院兽医研究所,江苏南京210014 [3]南京农业大学动物医学院,江苏南京210095 [4]南京军区军事医学研究所,江苏南京210002

出　　处：《中国预防兽医学报》2005年第3期171-174,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：江苏省自然科学基金资助项目(BK2003011)

摘　　要：将传染性法氏囊病病毒(IBDV)分离毒株ZB和TA3的细胞培养物采用不连续蔗糖密度梯度离心的方法浓缩纯化病毒,免疫BALB/c小鼠,运用淋巴细胞杂交瘤技术将免疫鼠脾细胞与SP2/0骨髓瘤细胞融合,建立了6株分泌抗IBDV单克隆抗体(McAb)的杂交瘤细胞系1C1、1E6、2B2、2G8、3A2、3E2。间接ELISA测定,杂交瘤细胞培养上清液的抗体效价为102,诱生腹水的抗体效价为106～107。相加ELISA分析,这6株单克隆抗体对应不同的病毒抗原表位。中和试验结果表明,2G8对病毒有较强的中和能力,腹水的中和效价为105。用2B1和3E2配对建立的夹心ELISA可特异地检测IB DV。The isolates ZB and TA3 of infectious bursal disease virus (IBDV) were propagated in chicken embryo fibroblasts (CEF) and purified by sucrose discontinuous gradient ultracentrifugation. Six hybridomas 1C_1,1E_6,2B_2,2G_8,3A_2 and 3E_2 secreting monoclonal antibodies (McAb) to IBDV were established by the fusion of mouse myeloma cells SP2/0 and spleen cells from BALB/c mice immunized with the purified IBDV. The antibody titers measured with indirect ELISA were 10~2 in culture supernatants and 10~6～10~7 in ascitic fluids.Additivity ELISA revealed that the six McAb recognized spatially independent epitopes.Neutralization test indicated that 2G_8 had strong neutralizing ability to IBDV,and the neutralization titer of ascitic fluid was 10~5.Sandwich ELISA for the specific detection of IBDV was developed utilizing 2B_1 and 3E_2.

关 键 词：传染性法氏囊病病毒 单克隆抗体 鉴定 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]