title: Antigenic analysis of grass carp reovirus using single-chain variable fragment antibody against IgM from Ctenopharyngodon idella.
abstract: Grass carp (Ctenopharyngodon idella) is an important species of freshwater aquaculture fish in China. However, grass carp reovirus (GCRV) can cause fatal hemorrhagic disease in yearling populations. Until now, a strategy to define the antigenic capacity of the virus's structural proteins for preparing an effective vaccine has not been available. In this study, some single-chain variable fragment antibodies (scFv), which could specifically recognize grass carp IgM, were selected from a constructed mouse naïve antibody phage display cDNA library. The identified scFv C1B3 clone was shown to possess relatively higher specific binding activity to grass carp IgM. Furthermore, ELISA analysis indicated that the IgM level in serum from virus-infected grass carp was more than two times higher than that of the control group at 5-7 days post infection. Moreover, Western blot analysis demonstrated that the outer capsid protein VP7 has a specific immuno-binding-reaction with the serum IgM from virus-infected grass carp. Our results suggest that VP7 can induce a stronger immune response in grass carp than the other GCRV structural proteins, which implies that VP7 protein could be used as a preferred immunogen for vaccine design.
authors: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China.
authors: CongLin
authors: C
authors: Chen
authors: XiaoYun
authors: X
authors: Sun
authors: LanJie
authors: L
authors: Liao
authors: ShaoXiang
authors: S
authors: Luo
authors: ZhouQuan
authors: Z
authors: Li
authors: XiaoHua
authors: X
authors: Zhang
authors: YaPing
authors: Y
authors: Wang
authors: QionLin
authors: Q
authors: Guo
authors: Qin
authors: Q
authors: Fang
authors: HePing
authors: H
authors: Dai
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000276:immunology; D002347:Carps / Q000276:immunology* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D005393:Fish Diseases / Q000097:blood / Q000276:immunology / Q000821:virology; D054884:Host-Pathogen Interactions / Q000276:immunology; D007075:Immunoglobulin M / Q000097:blood / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding / Q000276:immunology; D012087:Reoviridae / Q000276:immunology* / Q000502:physiology; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology*
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D007075:Immunoglobulin M; D019151:Peptide Library; D057127:Single-Chain Antibodies