癌变·畸变·突变 ›› 2009, Vol. 21 ›› Issue (2): 149-152.doi: 10.3969/j.issn.1004-616x.2009.02.017

• 技术与方法 • 上一篇    下一篇

基因免疫制备人Mig-2蛋白特异性单克隆抗体

胡元庆1,涂长春2,卫广森3,张宏权4   

  1. 1. 安徽科技学院 家禽疫病防控监测安徽省重点实验室，安徽　凤阳　233100；2.军事医学科学院军事兽医研究所，吉林　长春　130062；3.中国农业科学院兰州兽医研究所，甘肃　兰州　730046；4. Karolinska Institutet, Department of Microbiology, Pathology, and Immunology, SE-141 86 Huddinge, Sweden
  • 收稿日期:2008-06-30 修回日期:2008-09-16 出版日期:2009-03-30 发布日期:2009-03-30

Preparation of the Monoclonal Antibody Specific to Mig-2Protein by Genetic Immunization

HU Yuan-qing1,TU Chang-chun2,WEI Guang-sen3, ZHANG Hong-quan4,   

  1. 1. Anhui Science and Technology University, Anhui Key Laboratory of Poultry Epidemic Disease Prevention and Surveillance,Fengyang 233100,Anhui,China;2.Military Veterinary Research Institute, Academy of Military Medicine, Changchun 130062, Jilin,China;3. Lanzhou Veterinary Research Institute ,Chinese academy of Agricultural Sciences, Lanzhou 730046,Gansu,China;4.Karolinska Institutet, Department of Microbiology, Pathology, and Immunology, SE-141 86 Huddinge,Sweden
  • Received:2008-06-30 Revised:2008-09-16 Online:2009-03-30 Published:2009-03-30

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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。

关键词: 基因免疫, 单克隆抗体, Mig-2, 肿瘤治疗

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.

Key words: genetic immunization, monoclonal antibody, Mig-2, tumor therapy

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