抗传染性胰脏坏死病毒的重组单链抗体的流式细胞仪分选
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- 英文论文题名：Recombinant scFv antibodies against infectious pancreatic necrosis virus isolated by flow cytometry
- 论文作者：李渊 ; 徐黎明 ; 赵景壮 ; 刘淼 ; 卢彤岩
- 英文论文作者：Liyuan ; Li-Ming Xu ; Jing-Zhuang Zhao ; Miao Liu ; Tongyan-Lu ; Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences
- 年：2016
- 作者机构：中国水产科学研究院黑龙江水产研究所;
- 论文关键词：抗体库 ; 抗体抗原共表达 ; 细菌展示 ; 荧光激活细胞分选仪 ; 传染性胰脏坏死病毒chrtm213 ; 单链抗体
- 英文论文关键词：Antibody library ; Antibody-antigen co-expression ; Bacterial display ; fluorescence activated cell sorter ; Infectious pancreatic necrosis virus ChRtm213 ; Single chain variable fragment
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S943
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：547k
- 原文格式：D
- 会议级别：全国
摘要
传染性胰腺坏死病是中国养殖鲑鱼的常见疾病。在这项研究中,将虹鳟的单链抗体库(scFv)和中国传染性胰脏坏死病毒(IPNV)Ch Rtm213株的病毒蛋白VP2通过细菌展示技术共表达。然后通过流式细胞术(FCM)进行IPNV特异性抗体的筛选。经过三轮筛选,利用随机挑选单菌落的方法,获得了2株具有不同的荧光强度(MFI)的单链抗体克隆。将IPNV特异性的scFv抗体进行体外克隆、表达和纯化。结果显示纯化的抗体能够成功应用于Western印迹,酶联免疫吸附试验(ELISA)和荧光抗体试验(IFAT)。本研究提供了一种通过流式细胞仪高通量筛选抗体库的技术,通过该技术筛选获得的抗IPNV单链抗体可作为中国现行IPNV毒株的潜在的检测试剂。
Infectious pancreatic necrosis is a significant disease of farmed salmonids in China. In this study, a single chain variable fragment(scFv) antibody library derived from rainbow trout(Oncorhynchus mykiss) and viral protein VP2 of a Chinese infectious pancreatic necrosis virus(IPNV) isolate Ch Rtm213 were co-expressed by a bacterial display technology. The library was subjected to three rounds of screening by flow cytometry(FCM) to select IPNV specific antibodies. Two antibody clones with different mean fluorescence intensities(MFI) were obtained by picking colonies at random. The antibody clones were expressed and purified. The purified IPNV-specific scFv antibodies were used successfully in Western blotting, enzyme linked immunosorbent assay(ELISA) and an immunofluorescence antibody test(IFAT). This method provides a high throughput means to screen an antibody library by flow cytometry, and isolate a panel of antibody that can be used as potential reagents for the detection and study of IPNV that are prevalent in China.
Infectious pancreatic necrosis is a significant disease of farmed salmonids in China. In this study, a single chain variable fragment(scFv) antibody library derived from rainbow trout(Oncorhynchus mykiss) and viral protein VP2 of a Chinese infectious pancreatic necrosis virus(IPNV) isolate Ch Rtm213 were co-expressed by a bacterial display technology. The library was subjected to three rounds of screening by flow cytometry(FCM) to select IPNV specific antibodies. Two antibody clones with different mean fluorescence intensities(MFI) were obtained by picking colonies at random. The antibody clones were expressed and purified. The purified IPNV-specific scFv antibodies were used successfully in Western blotting, enzyme linked immunosorbent assay(ELISA) and an immunofluorescence antibody test(IFAT). This method provides a high throughput means to screen an antibody library by flow cytometry, and isolate a panel of antibody that can be used as potential reagents for the detection and study of IPNV that are prevalent in China.
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