抗犬瘟热病毒重组核衣壳蛋白单克隆抗体的制备及鉴定

以纯化的重组犬瘟热病毒(CDV)N蛋白免疫BALB/c小鼠,应用常规杂交瘤技术获得两株能稳定分泌特异性单克隆抗体的杂交瘤细胞系,分别命名为A_4C_6C_(12)和A_5B_8H_7。间接ELISA检测腹水效价分别为1:10~6、1:10~5;亚类鉴定结果分别为IgG2a、IgG2b,轻链均为κ型;Western blot和ELISA分析结果显示2株单克隆抗体均能与重组N蛋白和CDV发生反应,而与犬细小病毒及犬腺病毒等无交叉反应;ELISA叠加试验的增值结果表明两株单克隆抗体识别的抗原位点不同。特异性抗CDV-rN的单克隆抗体的获得,为进一步用于临床诊断研究奠定了基础。

Production and identification of monoclonal antibodies against recombinant nucleocapsid protein of canine distemper virus

Monoclonal antibodies(McAbs)against N protein of canine distemper virus were produced by fusing BALB/c mice myeloma cell line SP2/0 with B lymphocytes from spleen of BALB/c mice immunized with the purified recombinant N protein. Two hybridoma cell lines,A_4C_6C_(12)and A_5B_8H_7 that stably produce antibodies against N protein were identified by ELISA.The antibody titres of ascites for the hybridoma lines were 1:10~6 and 1:10~5,respectively.The subtypes of monoclonal antibodies were identified as IgG2a and IgG2b.Western blot and ELISA analysis showed that the McAbs specifically recognized certain antigenic epitopes of N protein of CDV,but not those of other reference viruses;Additive Index of ELISA competitive binding assay indicated that the two monoclonal antibodies were specific for different epitopes.