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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。
The murine anti-human DC-SIGN mAb was developed and the expression profile of DC-SIGN was analyzed using the obtained mAb. BALB / c mice were immunized with human DC-SIGN gene-transfected cells L929 / DC-SIGN. Spleen cells of immunized mice were fused with SP2 / 0 by B lymphocyte fusion technique. The tumor was repeatedly screened and cloned for several times. Western blot was used to detect the specific recognition of DC-SIGN. The subtypes and antigen recognition sites of the mAb were analyzed by rapid qualitative test strip and competition inhibition assay. Immunofluorescence analysis of its expression profile. Results A stable and stable hybridoma cell line secreting mouse anti-human DC-SIGN mAb was obtained. This mAb can specifically recognize human DC-SIGN molecule. The subtype of this antibody is IgG1, which is different from that of the commercial antibody E021819 The antigenic site of DC-SIGN is highly expressed in Mo-DC, which is a molecular marker of Mo-DC.