利用噬菌体展示技术淘选草鱼呼肠孤病毒的单链抗体  

ISOLATION OF SCFV ANTIBODIES AGAINST GCRV USING PHAGE DISPLAY TECHNOLOGY

作　　者：张凤[1,2] 李周全[1,2] 闫利明[2,3] 罗绍祥[1,2] 钟利桥[1,2] 张晓华[1] 袁丽[1] 方勤[3] 戴和平[1] 

机构地区：[1]中国科学院水生生物研究所淡水生态与生物技术国家重点实验室,武汉430072 [2]中国科学院大学,北京100049 [3]中国科学院病毒研究所病毒学国家重点实验室,武汉430071

出　　处：《水生生物学报》2014年第3期430-437,共8页Acta Hydrobiologica Sinica

基　　金：国家重点基础研究发展计划(批准号:2009CB118704)资助

摘　　要：草鱼呼肠孤病毒(GCRV)是引起我国大面积草鱼幼鱼出血病暴发的主要病原,其外衣壳蛋白VP5和VP7在病毒入侵宿主细胞过程中起着至关重要的作用。研究以原核表达的VP7、全长VP5、VP5的N端片段及C端片段为靶蛋白,利用已构建的噬菌体展示单链抗体文库进行淘选。经过3轮淘选后,共获得7个针对VP7、VP5、VP5N和VP5C的单链抗体。经过验证,识别原核表达的VP7的两个单链抗体能够成功识别天然GCRV病毒。此结果对于进一步研究GCRV与宿主细胞的相互作用机理奠定了基础。Grass carp reovirus（GCRV） is the main cause of severe outbreaks of hemorrhagic disease in fingerling and yearling grass carp.The virus outer capsid proteins,VP5 and VP7,have been shown to play critical roles in the invasion of host cells.In our study,prokaryotic-expressed VP7,VP5,and the N and C terminals of VP5 protein were used as targets to select the desired antibodies from phage-displayed scFv library.After 3 rounds of panning,we selected 7 antibodies against the prokaryotic-expressed antigens--VP7,VP5,VP5N and VP5C.Furthermore,we found that 2 antibodies against VP7 out of the total 7 were able to recognize native GCRV particles.Our study should provide an insight into the interactions between GCRV and its host cells.

关 键 词：草鱼呼肠孤病毒 VP5 VP7 噬菌体展示 单链抗体 

分 类 号：Q939.4[生物学—微生物学]