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国家自然科学基金(31072233)

作品数:2 被引量:9H指数:2
相关作者:张晓华陈丛琳方勤汪亚平廖兰杰更多>>
相关机构:中国科学院更多>>
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Antigenic analysis of grass carp reovirus using single-chain variable fragment antibody against IgM from Ctenopharyngodon idella被引量:6
2013年
Grass carp (Ctenopharyngodon idella) is an important species of freshwater aquaculture fish in China. However, grass carp reovirus (GCRV) can cause fatal hemorrhagic disease in yearling populations. Until now, a strategy to define the antigenic capacity of the virus's structural proteins for preparing an effective vaccine has not been available. In this study, some single-chain variable fragment antibodies (scFv), which could specifically recognize grass carp IgM, were selected from a constructed mouse na ve antibody phage display cDNA library. The identified scFv C1B3 clone was shown to possess relatively higher specific binding activity to grass carp IgM. Furthermore, ELISA analysis indicated that the IgM level in serum from virus-infected grass carp was more than two times higher than that of the control group at 5-7 days post infection. Moreover, Western blot analysis demonstrated that the outer capsid protein VP7 has a specific immuno-binding-reaction with the serum IgM from virus-infected grass carp. Our results suggest that VP7 can induce a stronger immune response in grass carp than the other GCRV structural proteins, which implies that VP7 protein could be used as a preferred immunogen for vaccine design.
CHEN CongLinSUN XiaoYunLIAO LanJieLUO ShaoXiangLI ZhouQuanZHANG XiaoHuaWANG YaPingGUO QionLinFANG QinDAI HePing
关键词:草鱼呼肠孤病毒抗原性分析可变区VP7基因CDNA文库
利用抗草鱼IgM的单链抗体分析草鱼呼肠孤病毒的免疫原性被引量:3
2012年
草鱼是中国重要的淡水养殖鱼种之一,草鱼呼肠孤病毒(GCRV)可引起一龄草鱼出现致命性的出血性疾病,严重影响草鱼养殖业的健康发展.目前,在草鱼中尚未建立一种有效的检测该病毒结构蛋白抗原性质的方法.本研究采用已构建的小鼠天然噬菌体展示抗体文库淘选得到可以特异性识别草鱼IgM的单链抗体,并在已鉴定的单链抗体阳性克隆中,筛选出对草鱼IgM有较好的特异性结合活性的单链抗体C1B3.ELISA分析表明,在感染草鱼出血病病毒5~7天后,感染组比对照组血清中IgM的含量增加了2倍多.进一步的Western blot结果显示,已感染病毒草鱼血清中的IgM能特异地识别草鱼的外壳蛋白VP7.结果表明,VP7相对于病毒的其他结构蛋白而言在草鱼中能产生更强的免疫反应,提示VP7应是用于疫苗设计的首选抗原.
陈丛琳孙小云廖兰杰罗绍祥李周全张晓华汪亚平郭琼林方勤戴和平
关键词:草鱼草鱼呼肠孤病毒免疫原性
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