中文摘要：

传染性囊病病毒（Infectious Bursal Disease Virus,IBDV）的非结构蛋白VP5在病毒感染细胞的不同阶段发挥着抑制或诱导细胞凋亡的重要作用。为制备抗血清I型IBDV VP5蛋白的单克隆抗体,取经His-VP5蛋白免疫4次的BALB/c小鼠的脾细胞与骨髓瘤细胞（SP2/0）进行融合,3次亚克隆后获得2株能稳定分泌VP5蛋白单克隆抗体的杂交瘤细胞株,分别命名为5EC7和2BE5。经间接ELISA测定,以上2株单抗的亲和力解离常数分别为1.14×10-10和4.87×10-10,亚类分别为Ig G2a和Ig G1。通过Western Blot和间接免疫荧光试验检测,2株单抗均能识别IBDV感染DF-1细胞后产生的VP5蛋白。Western Blot试验表明,2株单抗识别的抗原表位区域为VP5 N-端的110 aa。该单抗为研究血清I型IBDV VP5蛋白的生物学功能及病毒致病机理奠定基础。

英文摘要：

VP5 is a nonstructural protein of IBDV and plays an important role in inducing or inhibiting apoptosis during IBDV infection. In order to develop monoclonal antibodies（ Mc Abs） against IBDV VP5,we vaccinated BALB / c mice with His- VP5 protein.Hybridomas were generated by fusing mouse myeloma cells SP2 /0 with the splenocytes from the immunized mice. After screening and subcloning,we obtained 2 hybridoma cell lines（ 5EC7,and 2BE5） that stably secreted Mc Abs against the IBDV VP5. The dissociation constants（ Kds） of these Mc Abs were 1. 14 × 10-10 and 4. 87 × 10-10,respectively. The isotypes of these Mc Abs were Ig G2 a and Ig G1.These Mc Abs specially bound to VP5 in IBDV- infected DF- 1 cells as demonstrated by Western Blot and indirect immunofluorescence assay. The recognition regions of these Mc Abs were all located between 1to 10 aa of VP5 as demonstrated by Western Blot.These Mc Abs will be useful in analyzing the biological activities of VP5 and the mechanisms of IBDV infection.