【摘要】 研制鼠抗人DC-SIGN mAb,利用所获得的mAb对DC-SIGN的表达谱进行分析。以人DC-SIGN的基因转染细胞L929/DC-SIGN为免疫原免疫BALB/c小鼠;采用B淋巴细胞融合技术,将免疫小鼠脾脏细胞与SP2/0融合;经免疫荧光标记对杂交瘤进行反复筛选和多次的克隆化培养;利用Western blot检测抗体对DC-SIGN分子的特异性识别;采用快速定性试纸法及竞争抑制结合实验分析了该mAb的亚型及抗原识别位点;利用免疫荧光法对其表达谱进行了分析。结果获得了1株持续、稳定分泌鼠抗人DC-SIGN mAb的杂交瘤细胞株;该mAb能特异性识别人DC-SIGN分子;该抗体的亚型为IgG1,其与商品化抗体E021819识别不同的抗原位点;DC-SIGN分子特异性高表达于Mo-DC,是Mo-DC的标记分子。更多还原

【Abstract】 To prepare the mouse anti-human DC-SIGN monoclonal antibody and analyze the expression of the DC-SIGN molecules,BALB/c mice were immunized with L929/DC-SIGN transgenic cells,and the splenocytes of the mice were fused with SP2/0.The hybridoma cells were screened by immunofluorescence staining.The specific recognition of the mAb with DC-SIGN molecules was analyzed by Western blotting and Fast-strip analysis was performed to identify the sub-class of this mAb.The epitope recognized by this mAb was detected by competitive inhibition assay,and the expression of the DC-SIGN molecules was analyzed by immunofluorescence staining.In the end,one hybridoma cell line continuously and steadily secreting specific anti-human DC-SIGN mAb was obtained.This mAb specifically recognized DC-SIGN molecules.The mAb was IgG1 subclass and it could recognize specifically different antigenic epitopes just as the same as the commercial mAb E021819.DC-SIGN molecule primarily expresses on Mo-DC and serves as the marker of Mo-DC.更多还原