食管癌侵袭转移功能性抗体的制备  

Preparation of anti-human esophageal cancer invasion and metastasis functional monoclonal antibody

作　　者：遇珑[1] 胡海[1] 周转[1] 潘建[1] 杨治华[1] 冉宇靓[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤研究所分子肿瘤学国家重点实验室,北京100021

出　　处：《中国卫生检验杂志》2009年第4期765-768,共4页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金面上资助项目(30570818;30600279);国家重点基础研究发展规划973项目(2009CB521804)资助

摘　　要：目的:本研究旨在研制抗人食管癌侵袭转移功能性单克隆抗体,确定其抑制食管癌转移的特性。方法:用5例新鲜肺转移性食管癌原发瘤标本中分离的有核细胞免疫6只BALB/c小鼠。选择高血清效价小鼠脾细胞与S/P20细胞融合,采用HAT在甲基纤维素中选择培养。使用活细胞免疫荧光,迁移、侵袭、黏附、细胞增殖等方法筛选和鉴定有治疗价值的功能性单克隆抗体。结果:细胞融合挑选单个集落1260个。经活细胞免疫荧光筛选获得545株识别食管癌细胞膜蛋白的单抗。再通过免疫组化筛选获得了485株不与正常人食管组织反应的相对特异的单抗。通过对迁移、侵袭、黏附、增殖等功能筛选获得70株抑制迁移、32株抑制肺内皮黏附的相关单抗。结论:采用高通量制备、筛选、鉴定功能性单克隆抗体技术,获得了多株能够显著抑制食管癌细胞转移相关生物学行为和功能的单抗。Objective：The study was designed to manufacture anti - human esophageal cancer invasion and metastasis functional monoclonal antibody and determine their characteristics which inhibit metastasis of esophageal cancer. Methods：Nucleated cells were separated from 5 fresh primary tumor specimens of metastatic esophageal carcinoma to immunize 6 Balb/c mouse. The mouse of high serum titer was selected for the fusing of spleen cells with sp2/0 cells. The hybridoma cells were selected cultured with HAT in methyl cellulose. The therapeutic values of functional monoclonal antibody were screened by immigration and adhesion assay. Results： A total of 1260 monoclonal antibody clones were obtained by the fuse of spleen cells with sp2/0 cell. By living cells immunofluorescence screened, 545 clones were demonstrated to react with cell membrane protein of esophageal cancer. Monoclonal antibody 485 specially react with malignant esophageal tissues in immurohistochemistry assays through migration, adhesion, screening, and other functions of 70 clones were found to inhibit migration and 32 clones inhibiting intrapulmonary endothelia adhesions. Conclusion：We gained a number of monoclonal antibodies which could significantly inhibit of the metastatic behaviors of esophageal cancer cells by using of high - throughput screening and identification of functional monoclonal antibody library technology.

关 键 词：食管癌 转移:功能性抗体 

分 类 号：R735.1[医药卫生—肿瘤]