Selection and characterization of single-chain recombinant antibodies against spring viraemia of carp virus from mouse phage display library
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- 作者：Hong Liu ; Xiaocong Zheng ; Feng Zhang ; Li Yu ; Xiaohua Zhang ; Heping Dai ; Qunyi Hua ; Xiujie Shi ; Wensheng Lan ; Peng Jia ; Li Yuan ; Hong Liu
- 关键词：Phage display ; Single-chain fragment variable ; Spring viraemia of carp virus
- 刊名：Journal of Virological Methods
- 出版年：December, 2013
- 年：2013
- 卷：194
- 期：1-2
- 页码：178-184
- 全文大小：2325 K
文摘
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.