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Volume 22 Issue 1
February 2007
    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.
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    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
      Available online: 01 February 2007
    • 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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      1. Huang R,Chen X X,Zhang J H,et al. Experimental infection of red claw crayfish Cherax quadricarinatus with white spot Syndrome virus[J]. J Wuhan Univ(Nat Sci Ed),2004,50 (S2): 79-82.(in Chinee)

      2. Huang R,Xie Y L,Zhang J H,et al. A novel envelope protein involved in white spot syndrome virus infection[J]. J Gen Virol,2005,86: 1357-1361.
          doi: 10.1099/vir.0.80923-0

      3. Inouye K,Miwa S,Oseko N,et al. Mass mortalities of cultured Kuruma shrimp Penaeus japonicus in Japan in 1993: Electron microscopic evidence of the causative virus[J]. Fish Pathol,1994,29: 149-158.
          doi: 10.3147/jsfp.29.149

      4. Kanchanaphum P,Wongteerasupaya C,Sitidilokratana N,et al. Experimental transmission of white spot syndrome virus(WSSV) from crabs to shrimp Penaeus monodon [J]. Dis Aquat Organ,1998,34: 1-7.
          doi: 10.3354/dao034001

      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.
          doi: 10.1007/s00705-005-0719-2

      6. Liang Y,Huang J,Song X L,et al. Four viral protein of white spot syndrome virus (WSSV) that attach to shrimp cell membranes[J]. Dis Aquat Organ,2005,66: 81-85.
          doi: 10.3354/dao066081

      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.
          doi: 10.3354/dao027215

      9. Mayo M A. A summary of taxonomic changes recently approved by ICTV[J]. Arch Virol,2002,147: 1655-1656.
          doi: 10.1007/s007050200039

      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.
          doi: 10.1006/viro.1999.0088

      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.
          doi: 10.1006/viro.2001.1002

      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.

      14. Wang B L,Scopsi L,Martvig M,et al. Simplified purification and testing of colloidal gold probes[J]. Histochemistry,1985,83:109-115.
          doi: 10.1007/BF00495139

      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.
          doi: 10.1016/j.fsi.2003.09.006

      17. Wu W,Wang L,Zhang X. Identification of white spot syndrome virus(WSSV) envelope proteins involved in shrimp infection[J]. Virology,2005,332: 578-583.
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      18. Xie Y L,Zhang S Y,Huang R,et al. A modified technique for purifying white spot syndrome virus[J]. Virologica Sinica,2003,18 (4): 391-393. (in Chinese)

      19. Yang F,He J,Lin X,et al. Complete genome sequence of the shrimp white spot bacilliform virus [J]. J Virology,2001,75: 11811-11820.
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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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