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为制备犬瘟热病毒(CDV)单克隆抗体(MAb),本研究用腺病毒表达的CDV截短N蛋白(aa401~aa523)免疫BALB/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行融合并筛选到2株能够稳定分泌抗CDV N蛋白的MAb,命名为N1-C8和N1-C41。经间接ELISA测定N1-C8和N1-C41培养上清液及小鼠腹水效价分别为1∶1 024、1∶106和1∶512、1∶105。通过肽扫描的方法筛选出N1-C8的抗原表位为线性表位~(440)ENQGGDKYPIHFNDE454,但并未能够筛选出N1-C41的抗原表位,推测可能是因为其抗原表位为空间构象型。Western blot结果显示N1-C8和N1-C41两株MAb在识别CDV的不同毒力病毒株上有差异,可能与CDV强弱病毒株在N蛋白的差异变化有关。本研究两株MAb的制备为CDV不同毒力株的鉴别诊断提供了可能。
In order to prepare monoclonal antibodies against canine distemper virus (CDV), BALB / c mice were immunized with adenovirus expressing CDV truncated N protein (aa401 ~ aa523) and spleen cells immunized with SP2 / 0 Myeloma cells were fused and two strains of MAb that could stably secrete anti-CDV N protein were screened and named as N1-C8 and N1-C41. The indirect ELISA assay showed that the ascites titer of N1-C8 and N1-C41 culture supernatant and mouse were 1: 1 024,1:106 and 1: 512 respectively. The epitope of N1-C8 was screened by peptide scanning for the linear epitope of (440) ENQGGDKYPIHFNDE454, but the N1-C41 antigenic epitope was not able to be screened, presumably because of its spatial epitope . Western blot results showed that two MAb N1-C8 and N1-C41 were different in CDV-recognized virulent strains, which may be related to the differences in the N protein of CDV virulent strains. The preparation of two strains of MAb in this study provided the possibility of differential diagnosis of different virulent strains of CDV.