Xu Li-Ming, Li Tian-He, Zhou Bing, Guo Mo, Liu Miao, Zhao Jing-Zhuang, Cao Hong-Wei, Li De-Shan
Heilongjiang River Fishery Research Institute, Chinese Academy of Fishery Sciences, Harbin, 150070, People's Republic of China,
Biotechnol Lett. 2014 May;36(5):1029-35. doi: 10.1007/s10529-014-1463-z. Epub 2014 Feb 22.
Infectious bursal disease is an economically important disease that affects chickens worldwide. Here, a recombinant single chain variable fragment (scFv) antibody library derived from chickens immunized with VP2 protein of infectious bursal disease virus (IBDV) was constructed. The library was subjected to three rounds of screening by flow cytometry against VP2 protein through a bacteria display technology, resulting in the enrichment of scFv. Three scFv clones with different fluorescence intensity were obtained by random colony pick up. The isolated scFv antibodies were expressed and purified. Relative affinity assay showed the three clones had different sensitivity to VP2, in accordance with fluorescence activity cell sorting analysis. The potential use of the selected IBDV-specific scFv antibodies was demonstrated by the successful application of the isolated antibodies in western blotting assay and ELISA.
传染性法氏囊病是一种在全球范围内影响鸡群的具有重要经济意义的疾病。在此，构建了一个源自用传染性法氏囊病病毒（IBDV）的VP2蛋白免疫的鸡的重组单链可变片段（scFv）抗体文库。该文库通过细菌展示技术针对VP2蛋白进行了三轮流式细胞术筛选，从而实现了scFv的富集。通过随机挑取菌落获得了三个具有不同荧光强度的scFv克隆。对分离出的scFv抗体进行了表达和纯化。相对亲和力测定表明，这三个克隆对VP2具有不同的敏感性，这与荧光活性细胞分选分析结果一致。通过将分离出的抗体成功应用于蛋白质印迹分析和酶联免疫吸附测定，证明了所选的IBDV特异性scFv抗体的潜在用途。
