Abstract
VP2 is a structural protein of the foot-and-mouth disease virus (FMDV). In this study, a FMDV serotype-in-dependent monoclonal antibody (MAb), 4B2, was generated. By screening a phage-displayed random 12-peptide library, we found positive phages displaying the consensus motif ETTXLE (X is any amino acid (aa)), which is highly homologous to 6ETTLLE11 at the N-terminus of the VP2 protein. Subsequently, a series of GST-fusion proteins expressing a truncated N-terminus of VP2 were examined by western blot analysis using the MAb 4B2. The results indicated that the motif 6ETTLLE11 of VP2 may be the minimal requirement of the epitope recognized by 4B2. Moreover, a 12-aa peptide 2KKTEETTLLEDR13 was shown to be the minimal unit of the epitope with maximal binding activity to 4B2. Alanine-scanning analysis demonstrated thatThr7, Thr8, and Leu10 are the functional residues of the 4B2 epitope Glu6 and Leu9 are required residues, and Glu11 plays a crucial role in the binding of MAb 4B2. The fine mapping of the epitope indicated that MAb 4B2 has the potential to be used in FMDV diagnosis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bachrach, H.L. 1968. Foot-and-mouth disease. Annu. Rev. Microbiol. 22, 201–244.
Barnett, P.V., E.J. Ouldridge, D.J. Rowlands, F. Brown, and N.R. Parry. 1989. Neutralizing epitopes of type O foot-and-mouth disease virus. I. Identification and characterization of three functionally independent, conformational sites. J. Gen. Virol. 70(Pt 6), 1483–1491.
Barnett, P.V., A.R. Samuel, L. Pullen, D. Ansell, R.N. Butcher, and R.M. Parkhouse. 1998. Monoclonal antibodies, against O1 serotype foot-and-mouth disease virus, from a natural bovine host, recognize similar antigenic features to those defined by the mouse. J. Gen. Virol. 79(Pt 7), 1687–1697.
Chothia, C., A.M. Lesk, A. Tramontano, M. Levitt, S.J. Smith-Gill, G. Air, S. Sheriff, and et al. 1989. Conformations of immunoglobulin hypervariable regions. Nature 342, 877–883.
Curry, S., E. Fry, W. Blakemore, R. Abu-Ghazaleh, T. Jackson, A. King, S. Lea, J. Newman, and D. Stuart. 1997. Dissecting the roles of VP0 cleavage and RNA packaging in picornavirus capsid stabilization: the structure of empty capsids of foot-and-mouth disease virus. J. Virol. 71, 9743–9752.
Domingo, E., E. Baranowski, C. Escarmis, and F. Sobrino. 2002. Foot-and-mouth disease virus. Comp. Immunol. Microbiol. Infect. Dis. 25, 297–308.
Drake, J.W. and J.J. Holland. 1999. Mutation rates among RNA viruses. Proc. Natl. Acad. Sci. USA 96, 13910–13913.
Freiberg, B., B. Hohlich, B. Haas, A. Saalmuller, E. Pfaff, and O. Marquardt. 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.
Grubman, M.J. and B. Baxt. 2004. Foot-and-mouth disease. Clin. Microbiol. Rev. 17, 465–493.
Jackson, T., A.M. King, D.I. Stuart, and E. Fry. 2003. Structure and receptor binding. Virus Res. 91, 33–46.
Kitson, J.D., D. McCahon, and G.J. Belsham. 1990. Sequence analysis of monoclonal antibody resistant mutants of type O foot and mouth disease virus: evidence for the involvement of the three surface exposed capsid proteins in four antigenic sites. Virology 179, 26–34.
Lea, S., J. Hernandez, W. Blakemore, E. Brocchi, S. Curry, E. Domingo, E. Fry, and et al. 1994. The structure and antigenicity of a type C foot-and-mouth disease virus. Structure 2, 123–139.
Marquardt, O., M.M. Rahman, and B. Freiberg. 2000. Genetic and antigenic variance of foot-and-mouth disease virus type Asia1. Arch. Virol. 145, 149–157.
Muller, J.D., J.A. McEachern, K.N. Bossart, E. Hansson, M. Yu, A. Clavijo, J.M. Hammond, and L.F. Wang. 2008. Serotype-independent detection of foot-and-mouth disease virus. J. Virol. Methods 151, 146–153.
Saiz, J.C., M.J. Gonzalez, M.V. Borca, F. Sobrino, and D.M. Moore. 1991. Identification of neutralizing antigenic sites on VP1 and VP2 of type A5foot-and-mouth disease virus, defined by neutralization-resistant variants. J. Virol. 65, 2518–2524.
Thomas, A.A., R.J. Woortmeijer, W. Puijk, and S.J. Barteling. 1988. Antigenic sites on foot-and-mouth disease virus type A10. J. Virol. 62, 2782–2789.
Valarcher, J.F., N.J. Knowles, N.P. Ferris, D.J. Paton, V. Zakharov, A. Sherbakov, Y.J. Shang, and et al. 2005. Recent spread of FMD virus serotype Asia 1. Vet. Rec. 157, 30.
Wang, J.L., M.Q. Liu, J. Han, W.Z. Chen, W. Cong, G. Cheng, Y.H. Gao, and et al. 2007. A peptide of foot-and-mouth disease virus serotype Asia1 generating a neutralizing antibody response, and an immunostimulatory peptide. Vet. Microbiol. 125, 224–231.
Yang, M., A. Clavijo, R. Suarez-Banmann, and R. Avalo. 2007. Production and characterization of two serotype independent monoclonal antibodies against foot-and-mouth disease virus. Vet. Immunol. Immunopathol. 115, 126–134.
Author information
Authors and Affiliations
Corresponding author
Additional information
These authors contributed equally to this work.
Rights and permissions
About this article
Cite this article
Yu, Y., Wang, H., Zhao, L. et al. Fine mapping of a foot-and-mouth disease virus epitope recognized by serotype-independent monoclonal antibody 4B2. J Microbiol. 49, 94–101 (2011). https://doi.org/10.1007/s12275-011-0134-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-011-0134-1