新乡医学院学报 , Journal of Xinxiang Medical College, 编辑部邮箱 , 2011年06期 【作者】 倪亚萍; 王秦秦; 马丽菊; 李明军; 梁芳; 贾建桃; 马海智;
【Author】 NI Ya-ping1,WANG Qin-qin2,MA Li-ju2,LI Ming-jun1,LIANG Fang3,JIA Jian-tao4,MA Hai-zhi5(1.Department of Function,the Affiliated Hospital of Xi’an Medical College,Xi’an 710077,Shaanxi Province,China;2.Medical Experimental and Clinical Research Center,the First Affiliated Hospital of Kunming Medical College,Kunming 650032,Yunnan Province,China;3.Department of Oncology,Zhengzhou Central Hospital,Zhengzhou 450007,Henan Province,China;4.Department of Pathophysiology,Changzhi Medical College,Changzhi 046000,Shanxi Province,China;5.the Ophthalmology Hospital of Zhongshan University,Guangzhou 510080,Guangdong Province,China)
【机构】 西安医学院附属医院功能科; 昆明医学院第一附属医院临床医学实验研究中心; 郑州市中心医院肿瘤内科; 长治医学院病理生理学教研室; 中山医科大学眼科医院;
【摘要】 目的利用免疫亲和层析技术用自行制备的抗人肺癌单克隆抗体N35纯化其相应的肺癌相关抗原N35蛋白并对其进行鉴定。方法培养P35杂交瘤细胞,收集培养上清,利用Protein A-Sepharose CL-4B亲和层析法纯化单抗N35;用纯化得到的单抗与溴化氰活化的Protein A-Sepharose CL-4B制备免疫亲和柱;再将肺腺癌GLC-82细胞的裂解液通过此免疫亲和柱,纯化肺癌相关抗原N35;最后,对纯化得到的N35蛋白进行Western blot鉴定。结果应用Protein A-Sepharose CL-4B亲和层析法纯化得到纯度较高的单抗N35蛋白;应用溴化氰活化的Protein A-SepharoseCL-4B免疫亲和层析法纯化得到相对分子质量80 000的肺癌相关蛋白N35。结论纯化得到的肺癌相关蛋白N35是一种新的肿瘤相关抗原,可为肺癌的生物治疗提供新的靶点。 更多还原
【Abstract】 Objective To purify lung cancer-associated antigen N35(N35Ag) by immunoaffinity chromatography using our self-prepared monoclonal antibody N35(mAb N35).Methods After culturing P35 hybridoma cells and collecting their culture supernatant,the mAb N35 from the supernatant by affinity chromatography was purified by using Protein A-Sepharose CL-4B;the mAb N35 immunoaffinity column was made by using cyanogen bromide(CNBr) antivated Protein A-Sepharose CL-4B and purified mAb N35;N35Ag was purified by p... 更多还原 【关键词】 单克隆抗体; 肿瘤相关抗原; 蛋白纯化; 【Key words】 monoclonal antibody; lung cancer-associated antigen; purify; 【基金】 十五国家科技攻关计划资助项目(编号:2001B703b11)【分类号】R392.11
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