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Volume 27 Issue 5
October 2012

    Tong Lin, Junjun Shao, Huiyun Chang, Shandian Gao, Guozheng Cong and Junzheng Du. Generation of Monoclonal Antibodies against Non-structural Protein 3AB of Foot-and-Mouth Disease Virus[J]. Virologica Sinica, 2012, 27(5): 316-319. doi: 10.1007/s12250-012-3261-x
    Citation: Tong Lin, Junjun Shao, Huiyun Chang, Shandian Gao, Guozheng Cong, Junzheng Du. Generation of Monoclonal Antibodies against Non-structural Protein 3AB of Foot-and-Mouth Disease Virus .VIROLOGICA SINICA, 2012, 27(5) : 316-319.  http://dx.doi.org/10.1007/s12250-012-3261-x
    shu

    Generation of Monoclonal Antibodies against Non-structural Protein 3AB of Foot-and-Mouth Disease Virus

    cstr: 32224.14.s12250-012-3261-x

    • State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Vetersinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

    • 通讯作者： Huiyun Chang, changhuiyun@126.com; 
    • 收稿日期： 2012-05-31
      录用日期： 2012-09-12

    • To identify linear epitopes on the non-structural protein 3AB of foot-and-mouth disease virus (FMDV), BABL/c mice were immunized with the 3AB protein and splenocytes of BALB/c mice were fused with myeloma Sp2/0 cells. Two hybridoma monoclonal antibodies (mAbs) cell lines against the 3AB protein of foot-and-mouth disease virus (FMDV) were obtained, named C6 and E7 respectively . The microneutralization titer was 1:1024 for mAb C6, and 1:512 for E7. Both mAbs contain kappa light chains, and were of subclass IgG2b. In order to define the mAbs binding epitopes, the reactivity of these mAbs against FMDV were examined by indirect ELISA. The results showed that both mAbs can react with FMDV, but had no cross-reactivity with Swine Vesicular Disease (SVD) antigens. The titers in abdomen liquor were 1:5×106 for C6 and 1:2×106 for E7. In conclusion, the mAbs obtained from this study are specific for the detection of FMDV, can be used for etiological and immunological researches on FMDV, and have potential use in diagnosis and future vaccine designs.

    Generation of Monoclonal Antibodies against Non-structural Protein 3AB of Foot-and-Mouth Disease Virus

    • State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Vetersinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

    • Corresponding author: Huiyun Chang, changhuiyun@126.com
    • Received Date: 31 May 2012
      Accepted Date: 12 September 2012

      Fund Project: State Key Projects of Transgene Program 2011ZX08011-004State Key Projects of Transgene Program 2009ZX08006-002BState Key Projects of Transgene Program 2009ZX08007-008B

    • To identify linear epitopes on the non-structural protein 3AB of foot-and-mouth disease virus (FMDV), BABL/c mice were immunized with the 3AB protein and splenocytes of BALB/c mice were fused with myeloma Sp2/0 cells. Two hybridoma monoclonal antibodies (mAbs) cell lines against the 3AB protein of foot-and-mouth disease virus (FMDV) were obtained, named C6 and E7 respectively . The microneutralization titer was 1:1024 for mAb C6, and 1:512 for E7. Both mAbs contain kappa light chains, and were of subclass IgG2b. In order to define the mAbs binding epitopes, the reactivity of these mAbs against FMDV were examined by indirect ELISA. The results showed that both mAbs can react with FMDV, but had no cross-reactivity with Swine Vesicular Disease (SVD) antigens. The titers in abdomen liquor were 1:5×106 for C6 and 1:2×106 for E7. In conclusion, the mAbs obtained from this study are specific for the detection of FMDV, can be used for etiological and immunological researches on FMDV, and have potential use in diagnosis and future vaccine designs.
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      1. Barteling S J. 2004. Modern inactivated foot-and mouth disease vaccines: historical background and key elements in production and use. In: Foot-and-Mouth Disease: Current Perspectives, Sobrino F, Domingo E, ed. England: Horizon Bioscience, pp305-333.

      2. Freiberg B, Hohlich B, Haas B, et al. 2001. Type-independent detection of foot-and-mouth disease virus by monoclonal antibodies that bind to amino-terminal residues of capsid protein VP2. J Virol Methods, 92:199-205.
          doi: 10.1016/S0166-0934(00)00287-1

      3. Guo H, Liu Z, Sun S, et al. 2005. Immune response in guinea pigs vaccinated with DNA vaccine of foot-and-mouth disease virus O/China99.Vaccine, 23:3236-3242.
          doi: 10.1016/j.vaccine.2004.03.074

      4. Hema M, Nagendrakumar S B, Yamini R, et al. 2007. Chimeric tymovirus-like particles displaying foot-and-mouth disease virus non-structural protein epitopes and its use for detection of FMDV-NSP antibodies. Vaccine, 25: 4784-4794.
          doi: 10.1016/j.vaccine.2007.04.023

      5. Jackson T, King A M, Stuart D I, et al. 2003. Structure and receptor binding. Virus Res, 91:33-46.
          doi: 10.1016/S0168-1702(02)00258-7

      6. Lee H S, Heo E J, Jeoung H Y, et al. 2009. Enzyme-Linked Immunosorbent Assay Using Glycoprotein and Monoclonal Antibody for Detecting Antibodies to Vesicular Stomatitis Virus Serotype New Jersey. Clin Vaccine Immunol, 16(5):667-671.
          doi: 10.1128/CVI.00043-09

      7. Lin T, Du J Z, Shao J J, et al. 2009. Application of VP1 Protein to Develop Monoclonal Antibody against Foot-and-mouth Disease Virus Asia1 Type. Virol Sin, 24(3):215-220.
          doi: 10.1007/s12250-009-2986-z

      8. Lu Z J, Zhang X L, Fu Y F, et al.2010. Expression of the major epitope regions of 2C integrated with the 3AB non-structural protein of foot-and-mouth disease virus and its potential for differentiating infected from vaccinated animals. J Virol Methods, 170:128-133.
          doi: 10.1016/j.jviromet.2010.09.016

      9. Mohapatra J K, Pandey L K, Sanyal A, et al. 2011. Recombinant non-structural polyprotein 3AB-based serodiagnostic strategy for FMD surveillance in bovines irrespective of vaccination. J Virol Methods, 177(2) :184-192.
          doi: 10.1016/j.jviromet.2011.08.006

      10. Priyadharshini M L, Balamurugan V, Prabhudas K, et al. 2007. Expression of 3AB protein of foot and mouth disease virus in Pichia pastoris. Indian J Biotechnology, 6:329-335.

      11. Rosas M F, Vieira Y A, Postigo R, et al.2008. Susceptibility to viral infection is enhanced by stable expression of 3A or 3AB proteins from foot-and-mouth disease virus. Virology, 380:34-45.
          doi: 10.1016/j.virol.2008.06.040

      12. Shao J J, Chang H Y, Lin T, et al. 2011. Expression and utilization of 3AB nonstructural protein of foot-and-mouth disease virus in Escherichia coli. Chin J Biotechnol, 27(2): 180-184. (in Chinese)

      13. Smith D B. 2000. Generating fusions to glutathione S-transferase for protein studies. Methods Enzymol, 326:254-270.
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      14. Song M Y, Park S K, Kim C S, et al. 2008.Characterization of a novel anti-human TNF-α murine monoclonal antibody with high binding affinity and neutralizing activity. Exp Mol Med, 40(1):35-40.
          doi: 10.3858/emm.2008.40.1.35

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      Generation of Monoclonal Antibodies against Non-structural Protein 3AB of Foot-and-Mouth Disease Virus

        Corresponding author: Huiyun Chang, changhuiyun@126.com
      • State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Disease Reference Laboratory, Lanzhou Vetersinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
      • Received Date: 31 May 2012
      • Accepted Date: 12 September 2012
      Fund Project:  State Key Projects of Transgene Program 2011ZX08011-004State Key Projects of Transgene Program 2009ZX08006-002BState Key Projects of Transgene Program 2009ZX08007-008B

      Abstract: To identify linear epitopes on the non-structural protein 3AB of foot-and-mouth disease virus (FMDV), BABL/c mice were immunized with the 3AB protein and splenocytes of BALB/c mice were fused with myeloma Sp2/0 cells. Two hybridoma monoclonal antibodies (mAbs) cell lines against the 3AB protein of foot-and-mouth disease virus (FMDV) were obtained, named C6 and E7 respectively . The microneutralization titer was 1:1024 for mAb C6, and 1:512 for E7. Both mAbs contain kappa light chains, and were of subclass IgG2b. In order to define the mAbs binding epitopes, the reactivity of these mAbs against FMDV were examined by indirect ELISA. The results showed that both mAbs can react with FMDV, but had no cross-reactivity with Swine Vesicular Disease (SVD) antigens. The titers in abdomen liquor were 1:5×106 for C6 and 1:2×106 for E7. In conclusion, the mAbs obtained from this study are specific for the detection of FMDV, can be used for etiological and immunological researches on FMDV, and have potential use in diagnosis and future vaccine designs.

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