抗ICAM-5单克隆抗体的制备及实验研究

Preparation and experimental research of monoclonal antibodies against intercellular adhesion molecule-5

目的制备抗细胞间黏附分子5(intercellular adhesion molecule-5,ICAM-5)单克隆抗体(McAb)并进行实验研究。方法采用淋巴细胞杂交瘤技术制备小鼠源性单克隆抗体。采用ELISA法测定腹水ICAM-5McAb的效价,通过免疫荧光技术及Western blotting检测McAb的抗体特异性。结果获得3株分泌ICAM-5McAb的杂交瘤细胞株。ELISA检测效价可达1∶(4×105),免疫荧光与Western blotting显示3株McAb与ICAM-5分子具有良好的结合活性。结论成功制备了抗ICAM-5的McAb,为其进一步研究和应用奠定了基础。

Objective To study on the preparation and experimental research of monoclonal antibodies of intercel- lular adhesion molecule-5 （ICAM-5）. Methods Three hybridoma cell lines that steadily secret anti-ICAM-5 McAb were obtained by hybridoma technology. The titers were detected by ELISA. The specificity of antibodies were analyzed by Western blotting and immunofluorescence. Results Three hybridoma cell lines producing monoclonal antibodies against the ICAM-5 protein were obtained, ELISA showed that the titers of ascites were all more than l：（4x10~）. Three clones of MeAb were identified to have the highly specific affinity of activity to ICAM-5 protein by Western blotting and im- munofluorescence. Conclusion Three anti-ICAM-5 monoclonal antibodies has been obtained successfully.