流行性出血热病毒单克隆抗体的研究Study on the Monoclonal Antibody to Epidemic Haemorrhagic Fever Virus
朱青,俞永新
Zhu Qing National Institute for the Control of Pharmaceutical and Biological Product, Beijing
摘要(Abstract)：
应用流行性出血热病毒(EHFV)Z_(10)株制备的疫苗免疫BALB/c小鼠,取脾细胞与BALB/c小白鼠骨髓瘤SP2/0细胞融合,获得10株能分泌ENFV特异性单克隆抗体(McAb)的杂交瘤细胞,其McAbs小鼠腹水荧光抗体滴度达1:2000～1:51200。经免疫印迹试验及ELISA夹心法证明1 0株McAbs为抗EHFV核蛋白(NP)的McAb,均不具有中和抗体活性,有低滴度的血凝抑制活性;经间接免疫荧光试验(IFAT)证明,2株为组特异性的McAb,5株为亚组特异性的McAb,3株为型特异性的McAb。实验用冻存免疫脾细胞进行杂交获得成功,为杂交瘤细胞的研究提供了经验。
Balb/e mice were immunized with the vaccine prepared from the Z_0 strain of epidemic haemorrhagic fever virus(EHFV). 10 hybridoma cell strains which could secrete specific McAbs to EHFV were obtained by fusing the spleen cells of immunized Balb/c mice with murine myeloma SP 2/0 cells. The titers of MeAbs in murine ascites detected by immunofluorescence technique reached 1:2000~1:51200. Detecting by Western blot and ELISA sandwich method, the McAbs were proved to be specific to the nucleoprotein of EHFV, and showed low haemagglutination inhibition activity, but on neutralizing antibody activity was found; detecting by indirect fluorescent antibody technique, it demonstrated that, among the 10 cell strains, 2 couldsecrete group-specific McAbs, 5 could secrete subgroup-specific McAbs, and 3 could secrete type-specific McAbs. In addition, we also succeeded in hybridization using frozen immunized spleen cells, and provided an experience for the studies on hybridoma.
关键词(KeyWords)：
流行性出血热;单克隆抗体
Epidemic haemorrhagic fever McAb
基金项目(Foundation):
作者(Author):
朱青,俞永新
Zhu Qing National Institute for the Control of Pharmaceutical and Biological Product, Beijing
DOI: 10.13200/j.cjb.1993.02.3.zhuq.001
参考文献(References)：
- 1 宋干,等。流行性出血热防治手册,1987。
- 2 James W G. Monoclonal Antibodies: Principle and Practice. 1982．
- 3 MiChael F M. J Imm Meth, 1988, 114: 155．
- 4 Elliott L H, et al. J Gen Virol, 1984, 65: 1285．
- 5 俞永新,等。病毒学报,1988,4(2) :162。
- 6 汪力亚,等。免疫学杂志,1989,5(4) :213。
- 7 王灵枢,等。中华微生物学和免疫学杂志,1911,11:319。
- 8 徐志凯,等。病毒学报,1988,4(2) :113。
- 9 俞永新,等。中国生物制品学杂志,1992,5(3) ,97。