癌变·畸变·突变 ›› 2007, Vol. 19 ›› Issue (5): 347-350.doi: 10.3969/j.issn.1004-616x.2007.05.001

• 论著 •    下一篇

基因免疫制备人PAK4单克隆抗体及其部分特性的鉴定

胡元庆1，2/ 涂长春2,/ 卫广森3/ 张宏权4   

  1. (1.安徽科技学院动物科学学院，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 Institute, Department of Microbiology, Pathology and Immunology, SE_141 86 Huddinge, Sweden)
  • 收稿日期:2007-03-29 修回日期:2007-05-23 出版日期:2007-09-30 发布日期:2007-09-30
  • 通讯作者: 涂长春

Preparation and Partial Charac_terization of the Monoclonal Antibody Specific to PAK4

HU Yuan_qing1,2, TU Chang_chun2, WEI Guang_sen3, ZHANG Hong_quan4   

  1. (1.Animal Science College, Anhui Science and Technology University,Fengyang 233100，Anhui; 2.Military Veterinary Research Institute, Academy of Military Medicine, Changchun 130062, Jilin; 3.Lanzhou Veterinary Research Institute ,Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu;4.Karolinska Institute, Department of Microbiology, Pathology and Immunology, SE_141 86 Huddinge,Sweden)
  • Received:2007-03-29 Revised:2007-05-23 Online:2007-09-30 Published:2007-09-30
  • Contact: TU Chang_chun

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摘要： 背景与目的： 制备人PAK4单克隆抗体可以用于临床诊断恶性肿瘤，同时为深入探讨PAK4蛋白的功能提供可靠途径。 材料与方法： 用重组PAK4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PAK4特异性单抗。 结果： 得到稳定分泌PAK4抗体的单克隆杂交瘤细胞2 株，分别命名为2D1和3H4；间接ELISA方法测得2株PAK4单抗腹水效价为1∶1.0×104；单抗亚类鉴定表明2D1是IgG类，3H4是IgM类；免疫组化试验显示2 株单抗均与临床两例乳腺癌组织管状上皮细胞高度反应。 结论： PAK4单克隆抗体特异性较好,可用于临床肿瘤组织诊断。

关键词: 基因免疫, PAK4, 单克隆抗体, 免疫组化

Abstract: BACKGROUND ＆ AIM: For clinical diagnosis of malignant tumors and research the function of PAK4 protein, we prepared the McAbs of PAK4 protein. MATERIALS AND METHODS: The Balb/C mice were immunized with the recombinant plasmid of PAK4(the vector is P3XFLAG_CMV_10).By cell fusion and cell cloning, the McAbs of PAK4 were prepared. RESULTS: Two strains of hybridoma cells, producing anti_PAK4 antibody, named 2D1(PAK4) a subclass of IgG and 3H4(PAK4) a subclass of IgM, were obtained after fusion followed by three or four screening. The ascites of McAbs were prepared by injecting the hybridoma cells into the mice abdomen. Indirect ELISA results showed that the ascites of McAbs revealed high affinity with the PAK4 protein at the titer of 1∶1.0×104. Immunohistochemistry showed that the McAbs of PAK4 reacted specifically with the cells of breast cancer. CONCLUSION:These results demonstrated that the McAbs of PAK4 could be a specific marker for diagnosis of cancer.

Key words: gene immunology, monoclonal antibody, PAK4, immunohistochemistry

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