摘要
目的:制备特异性识别天然构象的人表皮生长因子受体-2(HER2)的单克隆抗体(mAb),用以指导Herceptin的临床应用。方法:利用纯化的原核表达的HER2融合蛋白作为抗原免疫BALB/c小鼠,取脾细胞与NS1骨髓瘤细胞按常规方法融合,ELISA法进行杂交瘤的初步筛选。构建HER2酵母展示系统,用酵母cellbase ELISA进行第2次筛选。将筛选到的阳性克隆株进行亚克隆获得稳定分泌细胞株,用免疫组织化学的方法鉴定其与SK-Br-3细胞和293a细胞的反应性。结果:通过ELISA筛选得到168株阳性杂交瘤,用酵母cellbase ELISA最终得到8株阳性杂交瘤,此8株免疫组化检测均与SK-Br-3细胞反应,而与293a细胞无反应。结论:成功制备了8株识别天然构象的HER2的mAb,能够用于免疫组化,为临床上使用Herceptin进行个性化治疗乳腺癌打下了基础。
AIM：To prepare and characterize the specific monoclonal antibodies （mAbs） that can recognize the native conformation of HER2 so as to guide the clinical application of Herceptin.METHODS：Hybridomas were generated by the fusion with the NS1 myeloma cell and spleen cell,which were from mice immunized by HER2 recombinant protein.After cell fusion,ELISA,Yeast cell base ELISA and immunohistochemistry were used to screening clones respectively.RESULTS：Eight clones that could react with SK-Br-3 and yeast displaying HER2 were selected from 168 clones which were positive in initial screening by ELISA.CONCLUSION：Successfully prepared 8 mAbs that can recognize the native conformation of HER2,which should provide useful reagent for personalized therapy against breast cancer with Herceptin.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2010年第5期456-458,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
天津市科技创新专项资金项目(08FDZDSH03000)
