小鼠Metrnl单克隆抗体的制备及鉴定

Preparation and characterization of monoclonal antibodies against mouse Metrnl

目的制备抗小鼠Metrnl单克隆抗体,并进行初步筛选鉴定。方法分别制备小鼠Metrnl多肽片段和全长蛋白作为免疫小鼠抗原,取免疫后小鼠脾细胞与SP2/0骨髓瘤细胞融合,筛选阳性杂交瘤细胞,并亚克隆获得稳定细胞株,制备腹水。用ELISA方法检测腹水抗体效价;用Western blot方法鉴定抗体。结果由14种多肽抗原制备的56株单克隆抗体中,未筛选出可用于Western blot识别Metrnl的抗体;由全长蛋白制备的25株抗体中,有12株可识别Metrnl蛋白。结论本实验成功制备了12株单抗,可用于识别检测小鼠Metrnl蛋白。

Objective To prepare monoclonal antibodies （mAb） against mouse Metrnl and identify its specificity. Methods Mouse Metrnl polypeptide fragments and full-length protein were prepared as antigens to immunize mice. Then mice spleen cells were fused with SP2/0 myeloma cells to obtain hybridoma cells which were screened for positive clone in order to subclone for stable cell lines. After ascites were prepared,ELISA method was used to detect the antibodies titer. Western blot method was applied to identify their specificity. Results No effective antibodies were identified from the ascites derived from 14 polypeptide antigens. Among the 25 antibodies derived from the full-length protein, 12 monoclonal antibodies can be used to identify the recombinant Metrnl protein. Conclusion 12 monoclonal antibodies were successfully prepared to identify mouse Metrnl protein.