用噬菌体蛋白Ⅸ展示抗HBsAg分子Fab段  

Generation of Fab-phage antibody using pⅨ phage display

作　　者：张达矜[1] 王琰[1] 化冰[1] 刘晓琳[1] 

机构地区：[1]海军总医院,北京100037

出　　处：《中华微生物学和免疫学杂志》2004年第1期52-56,共5页Chinese Journal of Microbiology and Immunology

基　　金：全军医药卫生十五重点项目 ( 0 1Z0 1 5)

摘　　要：目的　用丝状噬菌体外壳蛋白Ⅸ (pⅨ )展示抗体Fab段 ,并与外壳蛋白Ⅲ (pⅢ )展示效果进行比较。方法　采用PCR方法钓取噬菌体蛋白Ⅸ基因 ,构建pⅨ展示抗乙肝表面抗原Fab的表达载体 ,制备噬菌体抗体 ,鉴定其抗原结合活性和Fab呈现水平 ,观察了噬菌体洗脱回收效率 ,并与pⅢ展示进行比较。结果　噬菌体pⅨ可以展示有功能活性的抗体Fab段 ,但对Fab的展示率低于pⅢ ,展示率的高低可能与pⅨ展示的外源蛋白分子量大小有关。“结合 洗脱 回收”实验显示 ,酸洗脱法回收的Fab pⅢ噬菌体的感染活性大为下降 ,而对Fab pⅨ噬菌体无影响。结论　pⅨ成功展示Fab段 。Objective To display Fab on phage using pⅨ, and t o compare the Fab-pⅨ phage display with Fab-pⅢ phage display. Met hods Phage gene Ⅸ was obtained by PCR, and anti-HBsAg Fab-pⅨ exp ression vector was constructed. Phage antibody displaying Fab-pⅨ was prepared. Antigen binding activity and Fab displaying level were analyzed by ELISA. Behav ior of Fab-pⅨ and Fab-pⅢ phage display was analyzed by biopanning analysis a nd phage DNA semi-quantitation. Results Phage pⅨ could be used to display functional Fab. Fab displaying ratio of pⅨ display was lower than that of pⅢ display, which may be explained by the size of fusion protein. Biopanning analysis showed that the eluted Fab-pⅢ phage had dramatically decr eased viability compared with Fab-pⅨ phage. Conclusion We proved that pⅨ could be used to display Fab and pⅨ display had the advantag e of higher viability after elution than pⅢ though it may have lower Fab displa ying ratio.

关 键 词：噬菌体蛋白 抗乙肝表面抗原 基因表达载体 噬菌体抗体 噬菌体抗体库技术 

分 类 号：R392[医药卫生—免疫学]