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从鼠噬菌体展示文库中筛选和鉴定抗鲤鱼春病毒血症病毒的单链重组抗体。

Selection and characterization of single-chain recombinant antibodies against spring viraemia of carp virus from mouse phage display library.

机构信息

The Key Laboratory of Aquatic Animal Diseases, Shenzhen Exit & Entry Inspection and Quarantine Bureau, Shenzhen 518001, PR China; College of Fisheries, Huazhong Agricultural University, Wuhan 430070, PR China.

出版信息

J Virol Methods. 2013 Dec;194(1-2):178-84. doi: 10.1016/j.jviromet.2013.08.017. Epub 2013 Aug 29.

Abstract

Antibody-displaying phage library was selected after three rounds of panning against spring viraemia of carp virus (SVCV) by phage display technology. Eight positive clones which could produce soluble single-chain fragment variable (scFv) antibody induced by isopropyl-beta-d-thiogalactopyranoside (IPTG) were obtained. Dot blot results showed that the eight scFv antibodies could recognize SVCV. The soluble scFv antibodies showed a molecular weight 29 kD by Western blot. All scFv antibodies could recognize SVCV proteins specifically without cross-reaction with other virus proteins by ELISA. Indirect immunofluorescence results showed that all of these scFv antibodies reacted positively with virus in the SVCV-infected cells. These scFv antibodies will be useful tools to establish immunological detection methods for SVCV.

摘要

噬菌体展示技术经过三轮筛选,从鲤鱼春病毒血症病毒(SVCV)中获得了展示抗体的噬菌体文库。通过异丙基-β-D-硫代半乳糖苷(IPTG)诱导,获得了 8 个能够产生可溶性单链片段可变(scFv)抗体的阳性克隆。点印迹结果表明,这 8 个 scFv 抗体能够识别 SVCV。Western blot 结果显示,可溶性 scFv 抗体的分子量为 29 kD。ELISA 结果表明,所有 scFv 抗体都能特异性识别 SVCV 蛋白,与其他病毒蛋白无交叉反应。间接免疫荧光结果表明,这些 scFv 抗体均与 SVCV 感染细胞中的病毒呈阳性反应。这些 scFv 抗体将成为建立 SVCV 免疫检测方法的有用工具。

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