特异性抗人cyclin D1人源单链抗体的筛选  

Panning of CyclinD1-specific human single-chain fv from a phage antibody library

作　　者：李桂英[1] 朱迅[2] 周立宏[1] 邹德生[1] 郝东云[1] 田宇[3] 杜柏榕[2] 

机构地区：[1]吉林大学分子酶学工程教育部重点实验室,吉林长春130021 [2]吉林大学基础医学院免疫学教研室,吉林长春130021 [3]吉林大学中日联谊医院神经外科,吉林长春130033

出　　处：《中国实验诊断学》2006年第11期1248-1251,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金青年基金资助项目(30200256);吉林省科技发展计划资助项目(20050410-3)

摘　　要：目的从噬菌体抗体库中筛选人源性抗人cyclinD1蛋白的单链抗体并进行鉴定。方法以原核表达并用Ni-NTA Agarose纯化的重组人cyclinD1蛋白为抗原,通过“吸附-洗脱-扩增”淘选过程从噬菌体抗体库中筛选特异性抗人cyclinD1蛋白的单链抗体,通过ELISA方法对获得抗体的特异性和结合活性进行分析鉴定。结果经过4轮筛选,获得7株能与人cyclinD1蛋白结合的阳性克隆,其中3株噬菌体的可溶性表达单链抗体能与人cyclinD1蛋白有特异性结合活性。结论利用噬菌体抗体库技术可以不经免疫过程制备出特异性的人源性抗人cyclinD1抗体。Objective To obtain human phage single-chain antibodies （seFv） against human eyclinD1 from a phage antibody library. Methods Panning of the phage antibody library against human cyclinD1 antigen expressed in E. coli and purified with Ni-NTA agarose was conducted to select specific seFv. The binding activity and specificity of the isolated phage antibodies were confirmed by ELISA. Results After 4 rounds of panning, ＂adhension-amplification-elution＂ ,7 clones of seFv which could bind specifically to human cyclinD1 were obtained, but only 3 clones had binding activity after being expressed in soluble form.Conclusion It is feasible to isolate human anti-cyclinD1 phage antibodies from phage antibody library by-passing immunization.

关 键 词：人cyclinD1蛋白 噬菌体抗体库 单链抗体 淘选 

分 类 号：R392[医药卫生—免疫学]