摘要
用纯化的重组鸭IL-2蛋白免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,对杂交瘤细胞进行筛选,阳性孔经三次有限稀释法克隆,成功获得5株能稳定传代并分泌抗鸭IL-2单克隆抗体(mAb)的杂交瘤细胞,分别命名为:1H4、2B3、4G12、5F6、5H6,其中一株对鸡IL-2具有交叉反应性.各单克隆抗体腹水间接ELISA效价在1∶32000～1∶512000之间,抗体类型除mAb5H6为IgG2a亚类外,其余4株mAb均为IgG1亚类,5株mAb的轻链都为κ链.经Western-blot分析表明,所获得的5株抗鸭IL-2单克隆抗体与鸭IL-2具有反应性,推测它们是针对序列决定簇的.
Five hybridoma cell lines secreting monoclonal antibody(mAb) against duck IL-2 were produced by fusing mouse myeloma cells(SP2/0) with spleen cells from BALB/c immunized with the purified recombinant duck IL-2 protein and named as 1H4,2B3,4G12,5F6 and 5H6. One of the five hybridoma cell lines (5H6) could also react with chicken IL-2. The titres of ascitic fluids of the five mAbs ranged from 1∶32000 to 1∶512000 by indirect ELISA. Among the five mAbs, 5H6 belongs to IgG2a, and the other mAbs were of the same isotype of murine antibodies IgG1, according to the Mouse Immunoglobulin Standard Panel. The five murine antibodies' light chain hold the same isotype, κ. Western-blot analysis showed that the five mAbs could react with duck IL-2, they are supposed to against continuous determinants of duck IL-2.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2004年第2期169-172,共4页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
浙江省自然科学基金青年人才培养专项资助项目(R303145).
