从噬菌体抗体库中筛选抗HIV-Tat蛋白单链抗体

Screening of Human Anti-HIV Tat scFv from A Phage Display Anti Body Library

目的从Tomlinson(I+J)噬菌体抗体库中筛选人源化抗HIV-Tat蛋白单链抗体(scFv)并进行鉴定。方法以HIV-Tat蛋白为抗原,通过"吸附-洗脱-扩增"过程从噬菌体抗体库中筛选特异性抗Tat蛋白单链抗体,对其进行基因序列测定,并检测其抗原结合活性。结果经过筛选,获得了1株能与Tat蛋白特异性结合的阳性克隆,经检索kabat数据库,发现其轻、重链可变区分别属于VκⅠ型、VHⅢ型。结论利用噬菌体抗体库技术可以不经过免疫制备出高特异性的人源化抗Tat蛋白单链抗体。

Objective To clone human anfi-H1V-Tat single-chain antibodies（scFv） from Tomlinson （ I ＋ J） phage antibody li- brary. Methods Panning of phage antibody library against HIV-Tat was conducted to select specific anfi-HIV-Tat scFv. The antigen binding activity and the variable genes of the selected antibodies were analyzed and identified, Results After the panning, 1 Positive clone were obtained, which could bind Tat specifically. DNA sequence analyses indicated that the variable region gene of the obtained clone belonged to the subgroups of immunoglobulin gene family, Conclusion By-passing immunization, the specific human anti-Tat scFv also can be prepared by using phage antibody library,