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