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作 者:胡海燕[1] 姜春英[2] 张岩[1] 耿昭[1] 卢翌[1] 王晓明[1] 张冰[1] 刘贤锡[1]
机构地区:[1]山东大学医学院医学分子生物学研究中心,山东济南250012 [2]山东中医药大学附属医院肛肠科,山东济南250011
出 处:《中国现代普通外科进展》2005年第6期339-341,共3页Chinese Journal of Current Advances in General Surgery
基 金:卫生部科研基金资助项目(98-1-173)
摘 要:目的:制备鸟氨酸脱羧酶(ODC)单克隆抗体并观察其在大肠癌检测中的意义。方法:以人ODC重组蛋白免疫小鼠,用杂交瘤细胞技术制备单克隆杂交瘤细胞,以ELISA和Westernblot法验证阳性单克隆细胞所产生的免疫球蛋白。免疫组化检测大肠癌组织中ODC的表达水平。结果:筛选出稳定分泌抗ODC单抗的细胞株,ELISA检测证明其所分泌单抗属于IgG2a型。免疫组化检测大肠癌组织中ODC的表达显示该单抗检测效果较好,大肠癌组织中ODC的表达水平明显高于正常组织(P<0.05)。结论:成功地制备出ODC单克隆抗体,为进一步研究ODC基因表达与大肠癌的发病机理及其诊断的关系打下了良好基础。Objective :To prepare anti-ODC mab and investigate the ODC gene expression in the colorectal carcinoma. Methods : The anti-ODC mAb was prepared with classical hybridoma technique and was used to determine the ODO expression in colon cancer tissues by immunohistochemical assay.Results : A cell line which could steadily secret anti-ODO mAb was selected through four times suboloning. Western blotting reconfirmed the mAb and ELISA showed the subtype which was IgG2a. Immunohistochemioal staining showed that the coloreotal carcinoma Cells expressed significantly higher level of ODO than the normal coloreotal muoosa(98.6±1.03% vs 5.26± 5%, P〈0.01). Conelusion :The anti-ODC mAb was prepared successfully. It was used in immunoohemistry which showed ODC protein was very high in coloreotal carcinoma.
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