中文摘要：

为制备CIAV VP2单克隆抗体,将原核表达的His-VP2融合蛋白作为免疫原免疫BALB/c小鼠.经4次免疫后,取小鼠脾细胞与骨髓瘤细胞（SP2/0）融合.将原核表达的GST-VP2融合蛋白作为包被抗原,利用间接ELISA筛选阳性克隆.阳性细胞株经3次亚克隆后,获得3株稳定分泌抗CIAV VP2蛋白的杂交瘤细胞克隆,分别命名为3D7、3B8、2D3.经间接ELISA测定,以上3株杂交瘤细胞腹水效价分别为1∶8.0x 105、1∶4.0x105,以及1∶6.4x106,亲和力解离常数分别为7.34x10-11、2.65x 10-11,以及2.98x10-11.IgG亚类分别为IgG1、IgG2b以及IgG2b.经Western Blot试验证明,3D7株单抗能特异地识别CIAV VP2蛋白,其识别的抗原表位区域为VP2 N-端的2040 aa.经IFA试验证明,3株单抗均能识别天然的CIAV VP2蛋白.此单抗为研究CIAV VP2的生物学功能和CIAV致病机理奠定基础.

英文摘要：

In order to develop monoclonal antibodies （MeAbs）against CIAV VP2, we vaccinated BALB/c mice with His-VP2 protein.An indirect ELISA using GST-VP2 protein as coating antigen was established to screen positive clones. We obtained 3 hybridoma cellclones that stably secreted McAbs against VP2 and these clones were named 3D7,3B8 and 2D3, respectively.The titers in ascite fluidwere 1∶8.0＆#215; 105、1∶4.0x10^5 respectively.The dissociation constants （Kds）were 7.34x10-1, 2.65x10-11, 2.98x10-1, respectively.The isotypes were determined to be IgG1, IgG2b and IgG2b.The McAb 3D7 specifically bound to VP2 and its binding domain was locatedbetween 20 to 40 aa of VP2as demonstrated by Western Blot. These McAbs specifically recognized VP2 in CIAV-infected MDCC-MSBlceUs as examined by Indirect Fluorescence Antibody assay.These McAbs would help to analyze the biological activities of CIAVVP2 and elucidate the mechanism of CIAV infection.