兔出血热病毒单克隆抗体的研究

THE STUDY OF MONOCLONAL ANTIBODIES TO VIRUS OF ACUTE INFECTIOUS HAEMORRHAGIC DISEASE OF RABBIT(HDRV)

本文应用杂交瘤技术,将兔出血热病毒免疫的 BALB/C 小白鼠脾细胞与骨髓瘤细胞 SP_2/O 在PEG 存在下进行融合。杂交瘤细胞经 HAT 筛选,用间接 ELISA 法检测相应的阳性抗体。经三次克隆化及三个多月的传代扩大培养,获4株能稳定分泌单克隆抗体的杂交瘤细胞株,编号为84、83、44、19,其腹水滴度为1:1×10^(-5),1:1×10^(-4),1:1×10^(-5)及1:1×10^(-4)。经分类测定其免疫球蛋白均属于 IgG。亚类测定除44号属 IgG_(2a)外,其他均属 IgG_1。从阻断试验看,单抗的作用能被兔特异性抗血清所阻断,则证明84号腹水具有抗兔出血热病毒的特异性。应用 ELISA 法测定非标记抗体间对抗原的反应增值,说明83与44号间的反应增值很低,可能是识别同一个抗原决定簇,而84及19号则分别识别其他两个不同的抗原决定簇。同时也讨论了单克隆抗体发展的远景。

Four stable hybridoma cell lines No.84,83,44 and 19,which secreted monoclonal antibodies to the specific antigen (HDRV) are established and cloned from the fussion of murine mycloma cells (SP_2/O) and spleen cells of BALB/C mice immunized with HDRV.The four hybridoma cell clones successfully developed ascites in mice,the titers of antibodies in ascites were 1∶1×10^(-4)～1∶1×10^(-5),and serological classification was determined.McAb 84,83,19 were of IgG_1 subclass and McAb 44 was of IgG_(2a) subclass. According to the additivity index of reaction between two antibodies with one antigen,McAb 44,83 might recognize the same antigenic determinant, McAb 84,19 recognize other two determinants.