抗人肾癌G250单克隆抗体的制备及其功能鉴定  

Preparation and functional identification of anti-human-G250 monoclonal antibody

作　　者：朱海涛[1] 郑骏年[1] 李望[1] 郑宏祥[1] 温儒民[1] 刘晓筠[1] 刘俊杰[1] 裴冬生[1] 孔德领[2] 

机构地区：[1]江苏徐州医学院附属医院泌尿外科,221002 [2]南开大学生命科学院

出　　处：《中华实验外科杂志》2008年第10期1319-1320,共2页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30570385）;卫生部科学研究基金资助项目（WKJ2005-2-026）;江苏省内然科学基金资助项目（BK2007032）

摘　　要：目的大量制备、纯化抗人G250单克隆抗体（G250-McAb），并鉴定其与G250合成多肽及细胞天然抗原的结合特异性。方法将抗人G250杂交瘤细胞接种BALB／C小鼠腹腔，大量制备G250-McAb。采用正辛酸-硫酸铵沉淀法获得粗制抗体，然后经蛋白A亲和层析柱纯化抗体。采用荧光激活细胞分类术及免疫组织化学法鉴定G250-McAb与抗原结合特异性。结果成功制备抗人G250-McAb，腹水经纯化后，获得了高纯度抗人G250单克隆抗体，蛋白浓度达4．78g/L。荧光激活细胞分类术及免疫组织化学结果显示G250-McAb与G250表达阳性的Ketr-3细胞特异性结合，而不与G250表达阴性的ACHN细胞结合。结论成功制备并纯化G250-McAb，为进一步开展G250-McAb介导的肾癌诊断、治疗奠定了基础。Objective To prepare and purify anti-human-G250 monoelonal antibody （G250- McAb） and identify its function. Methods G250-McAb was generated in BALB/C mice abdominal cavity by injecting intraperitoneally with 1×10^6 hybridoma cells. The purity of McAb was determined by ammonium sulphate. The specificity of the McAb with G250 antigen was identified by fluorescence-activated cell sorting （FACS） and immunohischemistry. Results The McAb with high purity was obtained and the final concentration of McAb protein was about 4.78 g/L. It was also verified by FACS and immunohischemistry that McAb could bind to the surface of the G250 antigen positively expressed ceils such as Ketr-3 but not to the G250 antigen negatively expressed cells such as ACHN. Conclusion Anti-human-G250 McAb with high specificity was successfully generated and evaluated, which can be used in the diagnosis and treatment of renal cancer.

关 键 词：肾细胞癌 单克隆抗体 细胞结合 肾癌诊断 

分 类 号：R737.11[医药卫生—肿瘤] R392-33[医药卫生—临床医学]