【摘要】 背景与目的:Mig-2蛋白是肿瘤形成过程中的重要分子,本研究探讨制备人Mig-2蛋白的单克隆抗体以便为肿瘤诊断和治疗提供新途径。材料与方法:用重组Mig-2质粒(以P3XFLAG-CMV-10表达质粒为载体)免疫6～8周龄雌性BALB/c小鼠,通过细胞融合与克隆,筛选Mig-2特异性单抗。结果:得到稳定分泌Mig-2抗体的单克隆杂交瘤细胞3株,分别命名为3C4、4H2和1F8;间接ELISA方法测得3株Mig-2单抗腹水效价分别为1∶2.4×104、1∶3.6×104和1∶4.8×104,细胞分泌上清效价分别为1∶256、1∶32和1∶512;单抗亚类鉴定表明3株单抗均为IgM类。结论:制备的Mig-2单抗特异性和稳定性较好,可以用于肿瘤治疗相关研究。更多还原

【Abstract】 BACKGROUND AND AIM:Mig-2 protein is an important moleculer in the process of tumor formation,and is a target for tumor therapy.To search new effective ways to diagnose and treat malignant tumors,we prepared the McAbs of Mig-2 protein.MATERIALS AND METHODS:BALB/c mice were immunized with recombinant plasmid of Mig-2(the vector is P3XFLAG-CMV-10).By cell fusion and cell cloning,the McAbs of Mig-2 protein were prepared.RESULTS:Three strains of hybridoma cells 3C4,4H2 and 1F8 all secreting a subclass of IgM,anti-mig-2 antibody,were obtained after fusion followed by three or four screenings.The a scitic fluid McAbs were prepared by injecting the hybridoma cells into mice abdomen.The indirect ELISA results showed that ascitic McAbs revealed high affinity with the Mig-2 protein at the titer of 1∶2.4×104-1∶4.8×104,and the titer of cell supernatant was 1∶32-1∶512.CONCLUSION:Prepared McAbs of Mig-2 were specific and stable agent for tumor therapy research.更多还原