从小鼠噬菌体展示文库中筛选和鉴定抗传染性造血坏死病毒的单链重组抗体
Selection and characterization of single-chain recombinant antibodies against infectious haematopoietic necrosis virus from mouse phage display library.
作者信息
Liu Hong, Zheng Xiaocong, Shi Xiujie, Yu Li, Jia Peng, Wang Jinjin, He Junqiang, Lan Wensheng, Liu Hong, Wu Zhixin
机构信息
College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China.
Shenzhen Entry-Exit Inspection and Quarantine Bureau, Shenzhen 518045, China.
出版信息
J Virol Methods. 2014 Sep 1;205:61-7. doi: 10.1016/j.jviromet.2014.04.008. Epub 2014 Apr 18.
Six single-chain fragment variable (scFv) antibodies against infectious haematopoietic necrosis virus (IHNV) were selected from an antibody phage display library by phage display technology. The soluble scFv antibodies showed a molecular weight 32kDa by Western blot. Dot blot analysis revealed that the six scFv antibodies could recognize IHNV. For enzyme linked immunosorbent assay (ELISA), four scFv antibodies (P1A4, P1A12, P1D5 and P3E2) showed cross-reactivity with spring viraemia of carp virus (SVCV). However, none of the six scFv antibodies had cross-reaction with Pike fry rhabdovirus (PFRV), Soft-shelled turtle iridovirus (STIV), viral haemorrhagic septicemia virus (VHSV), or viral nervous necrosis virus (VNNV). Indirect immunofluorescence results showed that all of these scFv antibodies reacted positively with virus in the IHNV-infected cells. These scFv antibodies will be useful in diagnostic test development and pathogenesis studies for IHNV.
通过噬菌体展示技术从抗体噬菌体展示文库中筛选出六种针对传染性造血器官坏死病毒（IHNV）的单链可变片段（scFv）抗体。经蛋白质免疫印迹法检测，可溶性scFv抗体的分子量为32kDa。斑点印迹分析表明，这六种scFv抗体能够识别IHNV。在酶联免疫吸附测定（ELISA）中，四种scFv抗体（P1A4、P1A12、P1D5和P3E2）与鲤春病毒血症病毒（SVCV）存在交叉反应。然而，这六种scFv抗体均未与梭鲈弹状病毒（PFRV）、中华鳖虹彩病毒（STIV）、病毒性出血性败血症病毒（VHSV）或病毒性神经坏死病毒（VNNV）发生交叉反应。间接免疫荧光结果显示，所有这些scFv抗体在IHNV感染的细胞中均与病毒呈阳性反应。这些scFv抗体将有助于开发针对IHNV的诊断测试及开展发病机制研究。
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