一株鼠抗人B7-DC单克隆抗体的研制及其生物学特性的初步鉴定  

Preparation of a Novel Mouse Anti-Human B7-DC Mab and Primary Study of Its Biological Characteristics

作　　者：王萃[1] 陈永井[1] 

机构地区：[1]苏州大学医学部,江苏苏州215007

出　　处：《中国血液流变学杂志》2017年第1期36-40,共5页Chinese Journal of Hemorheology

摘　　要：目的 研制鼠抗人B7-DC分子单克隆抗体并鉴定其生物学特性.方法 以高表达人B7-DC分子的L929/B7-DC基因转染细胞株为免疫原,常规免疫6～8周龄的雌性Balb/c小鼠;利用B淋巴细胞杂交瘤技术,将免疫小鼠脾脏细胞与SP2/0融合,制备鼠抗人B7-DC单克隆抗体,用L929/B7-DC和293T/B7-DC基因转染细胞为抗体筛选阳性细胞,经间接免疫荧光标记法对杂交瘤进行反复筛选和多次克隆化培养,采用快速定性试纸分析法鉴定单抗Ig亚类,利用竞争抑制实验分析该单抗识别的抗原表位;采用免疫荧光标记分析该单抗在DC和肿瘤细胞系上的表达情况.结果 成功获得一株持续、稳定分泌鼠抗人B7-DC单克隆抗体的杂交瘤细胞株（命名为2D2）,为IgG1型,与商品化单抗识别不同的抗原表位,该单抗能特异性地识别人B7-DC分子在部分肿瘤细胞株上的表达.结论 该抗人B7-DC单抗的获得和生物学特性的初步鉴定,为进一步研究B7-DC在DCs细胞及肿瘤细胞上的作用机制奠定物质基础.Objective To prepare anti-human B7-DC monoclonal antibody and characterize its biological activities. Methods To prepare a novel mouse anti-human B7-DC monoclonal antibody and study its biological characteristics, 6～8 weeks female Balb/c mice were immunized with L929/B7-DC cell line as immunogen, then the mice spleen B cells were fused with SP2/0. Hybridoma cells were screened with L929/B7-DC and 293T/B7-DC transfectant cell lines by FCM. Fast-strip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by the mAb was studied by anti-B7-DC-PE competitive assay. Expression of B7-DC on dendritic cells （DCs） and tumor cell lines was studied by FACS assay. Results One hybridoma cell line named 2D2 was obtained successfully, which shows a property of secreting mouse anti-human B7-DC monoclonal antibody continuously and steadily. Conclusion Generation and characterization of a novel mouse anti-human B7-DC mAb may benefit further exploring the B7-DC functions on DCs and tumor cells in the process of immune response and studying how important the role B7-DC plays in cancer immunology.

关 键 词：B7-DC 单克隆抗体 共刺激分子 

分 类 号：R730.3[医药卫生—肿瘤]