Preparation and identification of monoclonal antibody specific to the epithelial cell adhesion molecule / 肿瘤研究与临床

ABSTRACT

Objective To prepare and identify the monoclonal antibody (mAb) specific to epithelial cell adhesion molecule (EpCAM) and explore the function of the mAb.Methods The EpCAM antigen expressed by the prokaryotic expression systems was used to immunize the BALB/c mice,and then the splenic cells from the mice were fused with the Sp2/0 cells to produce hybridomas secreting specific mAb.The positive clones were screened by the ELISA.The western blot analysis was used to identify the reactivity of the mAb to the antigen.Then the immunohistochemistry staining was used to detect EpCAM expression in the 3 primary colorectal carcinoma tissues.Results Three mAb specific to EpCAM were obtained by ELISA tests.Western blot results indicated that these three kinds of antibodies could react to the EpCAM antigen,but no response to the GST tag.Immunohistochemical staining results identified that these mAb could give positive signals to the primary colorectal carcinoma tissues from patients.Conclusion Three mAb specific to EpCAM are obtained and identified,which contributes to the diagnosis and therapy of the carcinoma in the future.