抗HSPC238单克隆抗体的制备和初步鉴定

Preparation and Characterization of the Monoclonal Antibody Against HSPC238 Antigen

目的制备抗HSPC238的单克隆抗体,为HSPC238的功能研究奠定基础。方法以纯化的重组蛋白HSPC238为抗原,免疫BALB/c小鼠,运用杂交瘤技术制备HSPC238单克隆抗体,并用间接ELISA法和Western-blot法对单克隆抗体的特性进行鉴定。结果成功建立两株稳定分泌抗HSPC238的单克隆抗体杂交瘤细胞株,分别命名为E001和E002。ELISA检测抗HSPC238单克隆抗体的腹水效价为1∶12800和1∶25600。两株单克隆抗体的免疫球蛋白亚类均为IgG1。通过Western-blot实验证实,两株单克隆抗体均能特异性结合真核细胞内源性HSPC238蛋白。结论成功制备了两株效价高、特异性好的抗HSPC238单克隆抗体,制备的抗HSPC238单克隆抗体可用于HSPC238蛋白的鉴定,为HSPC238蛋白的生物学功能研究奠定了基础。

Objective To establish HSPC238 hybridoma cell lines and to characterize the activities of these monoclonal antibodies （McAbs）. Method BALB/c mice were immunized with recombinant HSPC238 protein. Hybridoma cell lines were established by using hybridoma technique and monoclonal antibodies were prepared. Specificity of the antibodies was evaluated by Western-blot and enzyme linked immunosorbent assay （ELISA）. Result Two stable hybridoma cell lines, E001 and E002, secreting IgG1 antibody against HSPC238 McAbs were obtained. Both E001 and E002 are HSPC238 specific. Using the ELISA method, the titer of the ascite fluid obtained from E001 and E002 was 1：12800 and 1：25600, respectively. Western blotting analysis confirmed that the two monoclonal antibodies detect human HSPC238. Conclusion High titer HSPC238 specific monoclonal antibodies were successfully prepared. The antibodies may be used in the future study of the HSPC238 protein.