Skip to main content

Advertisement

SpringerLink
Log in

Identification of a conserved linear epitope using a monoclonal antibody against non-structural protein 3B of foot-and-mouth disease virus

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Foot-and-mouth disease virus (FMDV) is a member of the family Picornaviridae that has caused severe economic losses in many countries of the world. Regular vaccinations have been effectively used to control foot-and-mouth disease (FMD) in countries where the disease is enzootic. Distinguishing between infected and vaccinated animals in herds after immunization is an important component of effective eradication strategies. Nonstructural protein (NSP) 3B of FMDV is part of a larger antigen that is used for this differential diagnosis. In this study, an FMDV serotype-independent monoclonal antibody (MAb) against NSP 3B, 5D12, was generated. Using western blot, it was revealed that MAb 5D12 binds to three fragments of 3B displaying the motifs G1PYAGPLERQKPLK14, K18LPQQEGPYAGPMER32 and V45KEGPYEGPVKKPVA59. The motif G1PYAGPLERQKPLK14 was chosen for further mapping. Different truncated motifs derived from the motif G1PYAGPLERQKPLK14 were expressed as GST-fusion constructs for western blot analysis. The results showed that the 5-aa peptide P2YAGP6 was the minimal epitope reactive to MAb 5D12. Subsequent alanine-scanning mutagenesis analysis revealed that Pro2, Gly5 and Pro6 were crucial for MAb 5D12 binding to P2YAGP6. Furthermore, through sequence alignment analysis, the epitope PxxGP recognized by 5D12 was found to be present not only in 3B-1 but also in 3B2 and 3B3 and was highly conserved in seven serotypes of FMDV strains. Western blot analysis also revealed that the peptide epitope could be recognized by sera from FMDV-infected pigs and cattle. Thus, the 5D12-recognized 3B epitope identified here provides theoretical support for the development of MAb 5D12 as a differential diagnosis reagent for FMDV infection.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Atassi MZ, Tarlowski DP, Paull JH (1970) Immunochemistry of sperm whale myoglobin. VII. Correlation of immunochemical cross-reaction of eight myoglobins with structural similarity and its dependence on conformation. Biochim Biophys Acta 221:623–635

    Article  CAS  PubMed  Google Scholar 

  2. Bentley L, Fehrsen J, Jordaan F, Huismans H, du Plessis DH (2000) Identification of antigenic regions on VP2 of African horsesickness virus serotype 3 by using phage-displayed epitope libraries. J Gen Virol 81:993–1000

    Article  CAS  PubMed  Google Scholar 

  3. Bergmann IE, Astudillo V, Malirat V, Neitzert E (1998) Serodiagnostic strategy for estimation of foot-and-mouth disease viral activity through highly sensitive immunoassays using bioengineered nonstructural proteins. Vet Q 20(Suppl 2):S6–S9

    Article  CAS  PubMed  Google Scholar 

  4. Bergmann IE, Malirat V, Neitzert E, Beck E, Panizzutti N, Sanchez C, Falczuk A (2000) Improvement of a serodiagnostic strategy for foot-and-mouth disease virus surveillance in cattle under systematic vaccination: a combined system of an indirect ELISA-3ABC with an enzyme-linked immunoelectrotransfer blot assay. Arch Virol 145:473–489

    Article  CAS  PubMed  Google Scholar 

  5. Brownlie J (2001) Strategic decisions to evaluate before implementing a vaccine programme in the face of a foot-and-mouth disease (FMD) outbreak. Vet Rec 148:358–360

    CAS  PubMed  Google Scholar 

  6. Clavijo A, Wright P, Kitching P (2004) Developments in diagnostic techniques for differentiating infection from vaccination in foot-and-mouth disease. Vet J 167:9–22

    Article  CAS  PubMed  Google Scholar 

  7. Dawe PS, Pinto AA (1978) Antibody responses to type-specific and “virus-infection-associated” antigens in cattle vaccinated with inactivated polyvalent foot and mouth disease virus in North Malawi. Br Vet J 134:504–511

    CAS  PubMed  Google Scholar 

  8. De Diego M, Brocchi E, Mackay D, De Simone F (1997) The non-structural polyprotein 3ABC of foot-and-mouth disease virus as a diagnostic antigen in ELISA to differentiate infected from vaccinated cattle. Arch Virol 142:2021–2033

    Article  PubMed  Google Scholar 

  9. Denisova GF, Denisov DA, Yeung J, Loeb MB, Diamond MS, Bramson JL (2008) A novel computer algorithm improves antibody epitope prediction using affinity-selected mimotopes: a case study using monoclonal antibodies against the West Nile virus E protein. Mol Immunol 46:125–134

    Article  CAS  PubMed  Google Scholar 

  10. Doel TR (2003) FMD vaccines. Virus Res 91:81–99

    Article  CAS  PubMed  Google Scholar 

  11. Du Plessis DH, Romito M, Jordaan F (1995) Identification of an antigenic peptide specific for bluetongue virus using phage display expression of NS1 sequences. Immunotechnology 1:221–230

    Article  PubMed  Google Scholar 

  12. Geysen HM, Rodda SJ, Mason TJ, Tribbick G, Schoofs PG (1987) Strategies for epitope analysis using peptide synthesis. J Immunol Methods 102:259–274

    Article  CAS  PubMed  Google Scholar 

  13. Grubman MJ, Baxt B (2004) Foot-and-mouth disease. Clin Microbiol Rev 17:465–493

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Hohlich BJ, Wiesmuller KH, Schlapp T, Haas B, Pfaff E, Saahmuller A (2003) Identification of foot-and-mouth disease virus-specific linear B-cell epitopes to differentiate between infected and vaccinated cattle. J Virol 77:8633–8639

    Article  PubMed Central  PubMed  Google Scholar 

  15. Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497

    Article  CAS  PubMed  Google Scholar 

  16. Levy R, Forsyth CM, LaPorte SL, Geren IN, Smith LA, Marks JD (2007) Fine and domain-level epitope mapping of botulinum neurotoxin type A neutralizing antibodies by yeast surface display. J Mol Biol 365:196–210

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Mackay DK (1998) Differentiating infection from vaccination in foot-and-mouth disease. Vet Q 20(Suppl 2):S2–S5

    Article  PubMed  Google Scholar 

  18. Mackay DK, Forsyth MA, Davies PR, Salt JS (1998) Antibody to the nonstructural proteins of foot-and-mouth disease virus in vaccinated animals exposed to infection. Vet Q 20(Suppl 2):S9–11

    Article  PubMed  Google Scholar 

  19. May C, Doody JF, Abdullah R, Balderes P, Xu X, Chen CP, Zhu Z, Shapiro L, Kussie P, Hicklin DJ, Liao F, Bohlen P (2005) Identification of a transiently exposed VE-cadherin epitope that allows for specific targeting of an antibody to the tumor neovasculature. Blood 105:4337–4344

    Article  CAS  PubMed  Google Scholar 

  20. McVicar JW, Sutmoller P (1970) Foot-and-mouth disease: the agar gel diffusion precipitin test for antibody to virus-infection-associated (via) antigen as a tool for epizootiologic surveys. Am J Epidemiol 92:273–278

    CAS  PubMed  Google Scholar 

  21. Meyer RF, Babcock GD, Newman JF, Burrage TG, Toohey K, Lubroth J, Brown F (1997) Baculovirus expressed 2C of foot-and-mouth disease virus has the potential for differentiating convalescent from vaccinated animals. J Virol Methods 65:33–43

    Article  CAS  PubMed  Google Scholar 

  22. Neitzert E, Beck E, de Mello PA, Gomes I, Bergmann IE (1991) Expression of the aphthovirus RNA polymerase gene in Escherichia coli and its use together with other bioengineered nonstructural antigens in detection of late persistent infections. Virology 184:799–804

    Article  CAS  PubMed  Google Scholar 

  23. Oem JK, Kye SJ, Lee KN, Park JH, Kim YJ, Song HJ, Yeh M (2005) Development of synthetic peptide ELISA based on nonstructural protein 2C of foot and mouth disease virus. J Vet Sci 6:317–325

    PubMed  Google Scholar 

  24. Oem JK, Chang BS, Joo HD, Yang MY, Kim GJ, Park JY, Ko YJ, Kim YJ, Park JH, Joo YS (2007) Development of an epitope-blocking-enzyme-linked immunosorbent assay to differentiate between animals infected with and vaccinated against foot-and-mouth disease virus. J Virol Methods 142:174–181

    Article  CAS  PubMed  Google Scholar 

  25. Paton DJ, de Clercq K, Greiner M, Dekker A, Brocchi E, Bergmann I, Sammin DJ, Gubbins S, Parida S (2006) Application of non-structural protein antibody tests in substantiating freedom from foot-and-mouth disease virus infection after emergency vaccination of cattle. Vaccine 24:6503–6512

    Article  CAS  PubMed  Google Scholar 

  26. Rodriguez A, Dopazo J, Saiz JC, Sobrino F (1994) Immunogenicity of non-structural proteins of foot-and-mouth disease virus: differences between infected and vaccinated swine. Arch Virol 136:123–131

    Article  CAS  PubMed  Google Scholar 

  27. Shen F, Chen PD, Walfield AM, Ye J, House J, Brown F, Wang CY (1999) Differentiation of convalescent animals from those vaccinated against foot-and-mouth disease by a peptide ELISA. Vaccine 17:3039–3049

    Article  CAS  PubMed  Google Scholar 

  28. Shen X, Parks RJ, Montefiori DC, Kirchherr JL, Keele BF, Decker JM, Blattner WA, Gao F, Weinhold KJ, Hicks CB, Greenberg ML, Hahn BH, Shaw GM, Haynes BF, Tomaras GD (2009) In vivo gp41 antibodies targeting the 2F5 monoclonal antibody epitope mediate human immunodeficiency virus type 1 neutralization breadth. J Virol 83:3617–3625

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Silberstein E, Kaplan G, Taboga O, Duffy S, Palma E (1997) Foot-and-mouth disease virus-infected but not vaccinated cattle develop antibodies against recombinant 3AB1 nonstructural protein. Arch Virol 142:795–805

    Article  CAS  PubMed  Google Scholar 

  30. Sorensen KJ, Hansen CM, Madsen ES, Madsen KG (1998) Blocking ELISAs using the FMDV non-structural proteins 3D, 3AB, and 3ABC produced in the baculovirus expression system. Vet Q 20:17–20

    Article  PubMed  Google Scholar 

  31. Sorensen KJ, Madsen KG, Madsen ES, Salt JS, Nqindi J, Mackay DK (1998) Differentiation of infection from vaccination in foot-and-mouth disease by the detection of antibodies to the non-structural proteins 3D, 3AB and 3ABC in ELISA using antigens expressed in baculovirus. Arch Virol 143:1461–1476

    Article  CAS  PubMed  Google Scholar 

  32. Sugimura T, Suzuki T, Chatchawanchonteera A, Sinuwonkwat P, Tsuda T, Murakami Y (2000) Application of latex beads agglutination test for the detection of the antibody against virus-infection-associated (VIA) antigen of foot-and-mouth disease (FMD) virus. J Vet Med Sci Jpn Soc Vet Sci 62:805–807

    Article  CAS  Google Scholar 

  33. Sukupolvi-Petty S, Austin SK, Engle M, Brien JD, Dowd KA, Williams KL, Johnson S, Rico-Hesse R, Harris E, Pierson TC, Fremont DH, Diamond MS (2010) Structure and function analysis of therapeutic monoclonal antibodies against dengue virus type 2. J Virol 84:9227–9239

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Sun T, Lu P, Wang X (2004) Localization of infection-related epitopes on the non-structural protein 3ABC of foot-and-mouth disease virus and the application of tandem epitopes. J Virol Methods 119:79–86

    Article  CAS  PubMed  Google Scholar 

  35. Tribbick G (2002) Multipin peptide libraries for antibody and receptor epitope screening and characterization. J Immunol Methods 267:27–35

    Article  CAS  PubMed  Google Scholar 

  36. Uddowla S, Hollister J, Pacheco JM, Rodriguez LL, Rieder E (2012) A safe foot-and-mouth disease vaccine platform with two negative markers for differentiating infected from vaccinated animals. J Virol 86:11675–11685

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  37. Wang H, Zhao L, Li W, Zhou G, Yu L (2011) Identification of a conformational epitope on the VP1 G-H Loop of type Asia1 foot-and-mouth disease virus defined by a protective monoclonal antibody. Vet Microbiol 148:189–199

    Article  CAS  PubMed  Google Scholar 

  38. Wang LF, Du Plessis DH, White JR, Hyatt AD, Eaton BT (1995) Use of a gene-targeted phage display random epitope library to map an antigenic determinant on the bluetongue virus outer capsid protein VP5. J Immunol Methods 178:1–12

    Article  CAS  PubMed  Google Scholar 

  39. Yang D, Zhang C, Zhao L, Zhou G, Wang H, Yu L (2011) Identification of a conserved linear epitope on the VP1 protein of serotype O foot-and-mouth disease virus by neutralising monoclonal antibody 8E8. Virus Res 155:291–299

    Article  CAS  PubMed  Google Scholar 

  40. Yang M, Clavijo A, Li M, Hole K, Holland H, Wang H, Deng MY (2007) Identification of a major antibody binding epitope in the non-structural protein 3D of foot-and-mouth disease virus in cattle and the development of a monoclonal antibody with diagnostic applications. J Immunol Methods 321:174–181

    Article  CAS  PubMed  Google Scholar 

  41. Yu Y, Wang H, Zhao L, Zhang C, Jiang Z, Yu L (2011) Fine mapping of a foot-and-mouth disease virus epitope recognized by serotype-independent monoclonal antibody 4B2. J Microbiol 49:94–101

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by a grant from the Key Project of Heilongjiang Provincial Science and Technology Program (No. GA06B202).

Author information

Authors and Affiliations

  1. State Key Laboratory of Veterinary Biotechnology, Division of Livestock Infectious Diseases, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 427 Maduan Street, Harbin, 150001, People’s Republic of China

    Chaosi Li, Weifeng Liang, Wenming Liu, Decheng Yang, Haiwei Wang, Guohui Zhou & Li Yu

  2. Institute of Veterinary Medicine, Xinjiang Academy of Animal Science, 151 Eastern Kelamayi Street, Ürümqi, 830000, People’s Republic of China

    Wenge Ma

Authors
  1. Chaosi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Weifeng Liang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wenming Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Decheng Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Haiwei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Wenge Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Guohui Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Li Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guohui Zhou.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, C., Liang, W., Liu, W. et al. Identification of a conserved linear epitope using a monoclonal antibody against non-structural protein 3B of foot-and-mouth disease virus. Arch Virol 161, 365–375 (2016). https://doi.org/10.1007/s00705-015-2667-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-015-2667-9

Keywords

Search

Navigation