整合素阻断剂AP25单克隆抗体的制备与鉴定

Preparation and identification of integrin inhibitor AP25 monoclonal antibody

目的制备整合素阻断剂AP25的单克隆抗体,鉴定其效价和特异性。方法从抗原免疫BALB/c小鼠分离免疫脾细胞,通过杂交瘤技术将免疫脾细胞与小鼠骨髓瘤细胞SP2/0融合,筛选杂交瘤细胞株并进行扩大培养。采用小鼠体内诱生法制备腹水,进行纯化后得到抗AP25单克隆抗体,用ELISA和Western blot鉴定其效价和特异性。结果 ELISA检测抗AP25单克隆抗体效价可达1∶2×106,Western blot鉴定抗体特异性良好。结论获得了能够高表达抗AP25单克隆抗体的杂交瘤细胞株;腹水制备抗体,纯化后得到效价高、特异性良好的AP25单克隆抗体。

Objective To prepare integrin inhibitor AP25 monoelonal antibody and identify its titer and specificity. Methods BALB/c mice were immunized by antigen. Immune spleen cells wereseparated and fused with mouse myeloma cells SP2/0 by hybridoma technique. The hybridoma cell line that could secrete anti-AP25 antibody was screened and cultured. The mice were induced to generateascites,which was then purified to obtain anti-AP25 monoclonal antibody. The titer and specificity of monoclonal antibody were detected by ELISA and Western blot, respectively. Results The titer ofAP25 monoclonal antibody was 1∶2×10^6 and the specificity was favourable. Conclusion The hybridoma cell line that can highly express anti-AP25 monoclonal antibody has been obtained. Thepurified anti-AP25 monoclonal antibody is with high titer and good specificity.