【作者】 李渊； 徐黎明； 赵景壮； 刘淼； 卢彤岩；

【Author】 Liyuan;Li-Ming Xu;Jing-Zhuang Zhao;Miao Liu;Tongyan-Lu;Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences;

【机构】 中国水产科学研究院黑龙江水产研究所；

【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。更多还原

【Abstract】 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.更多还原