噬菌体抗体库淘筛方法的比较研究  

Research on Panning Methods of Phage Antibody Library

作　　者：杜东霞[1] 吴剑[2] 李小曼[2] 朱振久[1] 张冉[1] 

机构地区：[1]湖南师范大学医学院,湖南长沙410006 [2]湖南师范大学生命科学学院,湖南长沙410006

出　　处：《现代生物医学进展》2009年第4期730-732,共3页Progress in Modern Biomedicine

基　　金：湖南省自然科学基金资助项目(05JJ30065);湖南省教育厅科研基金一般项目(06C504)

摘　　要：目的:比较噬菌体抗体库的淘筛方法,从而为得到亲和力高、特异性强的抗体克隆打下基础。方法:在包被好抗原的2个细胞培养瓶中加入噬菌体抗体库,一个采用先直接加入宿主菌感染后,再将培养瓶内残留结合物洗脱中和后感染的方法,另一个则采用先洗脱并中和的噬菌体感染宿主菌,与抗原结合的剩余的噬菌体再行直接感染,将2种方法所得到的感染菌和援救所得的噬菌体进行CFU和PFU测定。结果:从实验的数据中可以发现,采用第一种方法所得次级库的容量要大于后一种方法所得的初级库。结论:噬菌体抗体库的淘筛方法必须按照实验的具体情况进行选择,如果要求严谨度适中,则可以采用直接感染的方法淘筛。Objective： The aim of this study is to find out the optimal panning methods of phage antibody library. Methods： The phage antibody library was mixed and incubated with coated antigens in cell culture flasks. The phages combined in one flask were infected directly with E.coli XLl-blue host cells, and those in the other one were eluted by Gly/HCl. The eluted phages were then allowed to infect host cells. The CFU and PFU of the infected bacteria and phages rescued by M13KO7 were determined by two panning methods. Results： The CFU and PFU of phages infected directly with bacteria are larger than those eluted with GIy/HC1 firstly. Conclusions： Selection of panning methods should be based on the actual condition. We can pan the phage antibody library through direct infection with the host cells when the library capacity is not large enough.

关 键 词：噬菌体抗体库 淘筛 CFU PFU 亲和力 

分 类 号：R364.5[医药卫生—病理学] Q514.3[医药卫生—基础医学]