PD-L1 mAb的制备、鉴定及其特性的研究

目的制备抗程序化死亡配体-1(Programmed Death-Ligand 1，PD-L1)的mAb杂交瘤细胞系并对其分泌的抗体特性进行研究。方法用我室纯化的PD-L1／IgG4 Fc融合蛋白免疫Balb／c小鼠，取小鼠脾细胞与骨髓瘤细胞SP2／0进行融合，经筛选和克隆化后，建立杂交瘤细胞系。以Western-blot等方法研究抗体特性。结果得到3株能够稳定分泌抗体，效价高的杂交瘤细胞系2D10、1C3和1A6，Western-blot显示2D10、1C3和1A6能与PD-L1(Mr为5800)结合。结论得到杂交瘤细胞系稳定分泌的mAb，在抗病毒免疫、抗肿瘤反应和自身免疫性疾病的免疫诊断和／或治疗方面可能有广阔的应用前景。

Establishment and identification of anti-PD-L1 antibodies

Objective To establish the anti-programmed death-ligand 1 (anti-PD-L1) mAb and to study characteristics of the monoclonal antibodies. Methods Balb/c mice were immunized with PD-L1/IgG4 Fc fusion protein and the immunized spleen cells were fused with SP2/0 to raise hybridoma. The property of the antibody was studied by chromosome assay, affinity assay, antigen epitope identification, and Western-blot method. Results Three hybridoma cell lines named 2D10, 1C3, and 1A6 were established, which could secrete high quality mAb. Conclusion 2D10, 1C3, and 1A6 cells have all the characteristics of hybridoma. They may have a broad prospect in diagnose and treatment in the filed of anti-virus immunity, anti-tumor, and autoimmune disease.