抗鸡CD25单克隆抗体的制备与鉴定  

Production and identification of monoclonal antibodies to chCD25

作　　者：吴炜[1] 滕巧泱[2] 吴佳俊[2] 黄振宇[2] 周继勇[2] 

机构地区：[1]浙江大学医学院附属第一医院传染病研究所,浙江杭州310003 [2]浙江大学动物预防医学研究所,浙江杭州310029

出　　处：《浙江大学学报（农业与生命科学版）》2007年第5期479-483,共5页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金资助项目(30625030;30471289).

摘　　要：用纯化的鸡IL-2Rα(chCD25)重组蛋白免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,对杂交瘤细胞及时筛选,阳性孔经3次有限稀释法克隆,成功获得1E1,1G1,4H5,5E1,6A1,6C2,6C97株能稳定传代并分泌抗chCD25单克隆抗体的杂交瘤细胞.该7株单克隆抗体的腹水间接ELISA效价达106以上.除了1E1为IgG2b外,抗体类型均为IgG1,轻链皆为κ.经Western blot和免疫细胞化学分析表明,7株单克隆抗体皆与chCD25重组蛋白和真核表达蛋白EGFP-chCD25反应.抗chCD25单克隆抗体的获得为开展禽类分子免疫学研究提供了一种新的工具.　BALB/c mice were immunized with purified recombinant chCD25.Spleen lymphotes were fused with SP2/0 myeloma cells.Seven hybridoma cell lines（1E1,1G1,4H5,5E1,6A1,6C2,6C9）secreting monoclonal antibodies（McAb）were produced by three-time limited dilution method.The titers of ascetic fluids of the seven McAbs were up to 106 in indirect ELISA.Besides 1E1 belonging to IgG2b,κ,isotypes and subclasses of other McAbs belonged to IgG1,κ.The results of western bolt and immunocytochemistry showed that these McAbs can specifically react with recombinant chCD25 and EGFP-chCD25,separately.It is proved that all these McAbs provides a useful instrument to study the expression of chCD25 in vivo and vitro in future.

关 键 词：鸡 CD25 杂交瘤细胞 单克隆抗体 

分 类 号：Q78[生物学—分子生物学] S831[农业科学—畜牧学]