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.
Keywords: Foot-and-Mouth disease virus (FMDV), Monoclonal antibody (mAb), 3AB
Footnotes
Foundation items: State Key Projects of Transgene Program (No.2011ZX08011-004; 2009ZX08007-008B; 2009ZX08006-002B).
References
- 1.Barteling S. J. Modern inactivated foot-and mouth disease vaccines: historical background and key elements in production and use. In: Sobrino F., Domingo E., editors. Foot-and-Mouth Disease: Current Perspectives. England: Horizon Bioscience; 2004. pp. 305–333. [Google Scholar]
- 2.Freiberg B., Hohlich B., Haas B., et al. 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. 2001;92:199–205. doi: 10.1016/S0166-0934(00)00287-1. [DOI] [PubMed] [Google Scholar]
- 3.Guo H., Liu Z., Sun S., et al. Immune response in guinea pigs vaccinated with DNA vaccine of foot-and-mouth disease virus O/China99. Vaccine. 2005;23:3236–3242. doi: 10.1016/j.vaccine.2004.03.074. [DOI] [PubMed] [Google Scholar]
- 4.Hema M., Nagendrakumar S. B., Yamini R., et al. Chimeric tymovirus-like particles displaying foot-and-mouth disease virus non-structural protein epitopes and its use for detection of FMDV-NSP antibodies. Vaccine. 2007;25:4784–4794. doi: 10.1016/j.vaccine.2007.04.023. [DOI] [PubMed] [Google Scholar]
- 5.Jackson T., King A. M., Stuart D. I., et al. Structure and receptor binding. Virus Res. 2003;91:33–46. doi: 10.1016/S0168-1702(02)00258-7. [DOI] [PubMed] [Google Scholar]
- 6.Lee H. S., Heo E. J., Jeoung H. Y., et al. Enzyme-Linked Immunosorbent Assay Using Glycoprotein and Monoclonal Antibody for Detecting Antibodies to Vesicular Stomatitis Virus Serotype New Jersey. Clin Vaccine Immunol. 2009;16(5):667–671. doi: 10.1128/CVI.00043-09. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7.Lin T., Du J. Z., Shao J. J., et al. Application of VP1 Protein to Develop Monoclonal Antibody against Foot-and-mouth Disease Virus Asia1 Type. Virol Sin. 2009;24(3):215–220. doi: 10.1007/s12250-009-2986-z. [DOI] [Google Scholar]
- 8.Lu Z. J., Zhang X. L., Fu Y. F., et al. 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. 2010;170:128–133. doi: 10.1016/j.jviromet.2010.09.016. [DOI] [PubMed] [Google Scholar]
- 9.Mohapatra J. K., Pandey L. K., Sanyal A., et al. Recombinant non-structural polyprotein 3AB-based serodiagnostic strategy for FMD surveillance in bovines irrespective of vaccination. J Virol Methods. 2011;177(2):184–192. doi: 10.1016/j.jviromet.2011.08.006. [DOI] [PubMed] [Google Scholar]
- 10.Priyadharshini M. L., Balamurugan V., Prabhudas K., et al. Expression of 3AB protein of foot and mouth disease virus in Pichia pastoris. Indian J Biotechnology. 2007;6:329–335. [Google Scholar]
- 11.Rosas M. F., Vieira Y. A., Postigo R., et al. Susceptibility to viral infection is enhanced by stable expression of 3A or 3AB proteins from foot-and-mouth disease virus. Virology. 2008;380:34–45. doi: 10.1016/j.virol.2008.06.040. [DOI] [PubMed] [Google Scholar]
- 12.Shao J. J., Chang H. Y., Lin T., et al. Expression and utilization of 3AB nonstructural protein of foot-and-mouth disease virus in Escherichia coli. Chin J Biotechnol. 2011;27(2):180–184. [PubMed] [Google Scholar]
- 13.Smith D. B. Generating fusions to glutathione S-transferase for protein studies. Methods Enzymol. 2000;326:254–270. doi: 10.1016/S0076-6879(00)26059-X. [DOI] [PubMed] [Google Scholar]
- 14.Song M. Y., Park S. K., Kim C. S., et al. Characterization of a novel anti-human TNF-α murine monoclonal antibody with high binding affinity and neutralizing activity. Exp Mol Med. 2008;40(1):35–40. doi: 10.3858/emm.2008.40.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]