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Volume 22 Issue 1
February 2007

    Wan-gang GU, Jun-fa YUAN, Ge-lin XU, Li-juan LI, Ni LIU, Cong ZHANG, Jian-hong ZHANG and Zheng-li SHI. Production and Characterization of Monoclonal Antibodies of Shrimp White Spot Syndrome Virus Envelope Protein VP28[J]. Virologica Sinica, 2007, 22(1): 21-25.
    Citation: Wan-gang GU, Jun-fa YUAN, Ge-lin XU, Li-juan LI, Ni LIU, Cong ZHANG, Jian-hong ZHANG, Zheng-li SHI. Production and Characterization of Monoclonal Antibodies of Shrimp White Spot Syndrome Virus Envelope Protein VP28 .VIROLOGICA SINICA, 2007, 22(1) : 21-25.
    shu

    对虾白斑综合征病毒囊膜蛋白VP28单克隆抗体的制备 和特性研究

    • 1.

      中国科学院武汉病毒研究所 病毒学国家重点实验室,湖北武汉,430071;2卫生部生物制品研究所，湖北武汉，430060;3华中师范大学 生命科学学院，湖北武汉,430079

    • 通讯作者： 石正丽, zlshi@wh.iov.cn
    • 收稿日期： 2006-04-24
      录用日期： 2006-09-07
    • 摘要：用纯化的对虾白斑综合征病毒（White spot syndrome virus, WSSV）免疫BALB/c小鼠，用大肠杆菌表达的VP28蛋白通过ELISA方法筛选到六株抗囊膜蛋白VP28的单抗细胞株。用这6株单抗体外中和病毒后进行感染实验，结果表明，其中的4株单抗对病毒有中和作用，可以明显延缓螯虾被病毒感染后的起始死亡时间。Western blot杂交显示，所有单抗均为针对VP28空间结构的抗体。挑选其中的一株单抗7B4，用胶体金进行标记可以将VP28定位在病毒囊膜上。这些单克隆抗体可以用于建立WSSV的诊断技术。

    Production and Characterization of Monoclonal Antibodies of Shrimp White Spot Syndrome Virus Envelope Protein VP28

    • 1.

      State Key Laboratory of Virology,Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan 430071,China

    • 2.

      Wuhan Institute of Biological Products,Ministry of Health,Wuhan 430060,China

    • 3.

      College of Life Science,Central China Normal University,Wuhan 430079,China

    • Corresponding author: Zheng-li SHI, zlshi@wh.iov.cn
    • Received Date: 24 April 2006
      Accepted Date: 07 September 2006
    • The BALB/c mice were immunized with purified White Spot Syndrome Virus (WSSV). Six monoclonal antibody were selected, by ELISA, with VP28 protein expressed in E. Coli. The in vitro neutralization experiments showed that 4 of them could inhibit the virus infection to crayfish. Western-blot revealed that all these monoclonal antibodies were against the nature structure of VP28. The monoclonal antibody 7B4 was labeled with colloidal gold particles and used to locate the VP28 on virus envelope by immunogold labelling. These monoclonal antibodies could be used to develop immunological diagnosis methods of WSSV infection.
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      5. Li L J,Yuan J F,Cai C A,et al. Multiple envelope proteins are involved in white spot syndrome virus(WSSV) infection in crayfish[J]. Arch Virol,2006,151:1309-1317.
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      7. Liu W,Wang Y T,Tian D S,et al. Detection of white spot syndrome virus(WSSV) of shrimp by means of monoclonal antibodies (MAbs) specific to an envelope protein(28 kDa) [J]. Dis Aquat Organ,2002,49 (1): 11-18.

      8. Lo C F,Ho C H,Peng S E,et al. White spot syndrome baculovirus (WSBV) detected in cultured and captured shrimp,crabs and other arthropods[J]. Dis Aquat Organ,1996,27: 215-225.
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      10. Tsai J M,Wang H C,Leu J H,et al. Identification of the nucleocapsid,tegument,and envelope proteins of the shrimp white spot syndrome virus virion[J]. J Virol,2006,80: 3021-3029.
          doi: 10.1128/JVI.80.6.3021-3029.2006

      11. van Hulten M C,Westenberg M,Goodall S D,et al. Identification of two major virion protein genes of white spot syndrome virus of shrimp[J]. Virology,2000,266: 227-236.
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      12. van Hulten M C,Witteveldt J,Peters S,et al. 2001a. The white spot syndrome virus DNA genome sequence[J]. Viro-logy,286: 7-22.
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      13. van Hulten M C,Witteveldt J,Snippe M,et al. 2001b. White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp[J]. Virology,285: 228-233.

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      15. Wei K Q,Xu Z R. Effect of white spot syndrome virus envelope protein Vp28 expressed in silkworm-(Bombyx mori) pupae on disease resistance in Procam-buras clarkii[J]. Acta Biologiae Experimentalis Sinica,2005,38:190-198.(in Chenese)

      16. Witteveldt J,Vlak J M,van Hulten M C. Protection of Penaeus monodon against white spot syndrome virus using a WSSV subunit vaccine[J]. Fish Shellfish Immunol,2004,16: 571-579.
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      Production and Characterization of Monoclonal Antibodies of Shrimp White Spot Syndrome Virus Envelope Protein VP28

        Corresponding author: Zheng-li SHI, zlshi@wh.iov.cn
      • 1. State Key Laboratory of Virology,Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan 430071,China
      • 2. Wuhan Institute of Biological Products,Ministry of Health,Wuhan 430060,China
      • 3. College of Life Science,Central China Normal University,Wuhan 430079,China
      • Received Date: 24 April 2006
      • Accepted Date: 07 September 2006

      Abstract: The BALB/c mice were immunized with purified White Spot Syndrome Virus (WSSV). Six monoclonal antibody were selected, by ELISA, with VP28 protein expressed in E. Coli. The in vitro neutralization experiments showed that 4 of them could inhibit the virus infection to crayfish. Western-blot revealed that all these monoclonal antibodies were against the nature structure of VP28. The monoclonal antibody 7B4 was labeled with colloidal gold particles and used to locate the VP28 on virus envelope by immunogold labelling. These monoclonal antibodies could be used to develop immunological diagnosis methods of WSSV infection.

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