Abstract
Foot-and-mouth disease virus (FMDV) has led to serious losses in the farming industry worldwide, particularly in cattle and swine. In developing countries, the control and eradication of FMD rely upon vaccination, in which the inactivated vaccine is predominant. In the preparation of inactivated vaccine, a series of purification methods were used to remove non-structural proteins (NSPs). It is necessary to develop a quantitative detection method of residual NSP and confirm a threshold value for the evaluation of the vaccine. Meanwhile, it is also important to develop a sensitive and rapid diagnostic method to distinguish infected animals from vaccinated animals (DIVA). In this study, three monoclonal antibodies (MAbs) against NSP 3ABC, designated 2G5, 9E2, and 1E10, were used. Subsequently, a series of overlapping peptides were expressed using a prokaryotic expression system to determine the minimal epitopes identified by the MAbs. Three linear B cell epitopes (BCEs), “92EYIEKA97” “23EGPYAGPLE31” and “209EPHH212”, were identified by MAbs 2G5, 9E2, and 1E10, respectively. Alanine-scanning mutagenesis analysis confirmed the critical amino acid in these epitopes. The epitope “92EYIEKA97” is located in 3A, which is deleted in some natural deletion mutants that result in a change in virus tropism. MAb 9E2 that identified the epitope “23EGPYAGPLE31” reacted with 3B1 and 3B2, but did not react with 3B3. In combination with sequence alignment analysis, the epitope “23EGPYAGPLE31” is highly conserved among different FMDV isolates. Preliminary screening using the known positive and negative sera indicated the MAb 9E2 has the potential for the development of a diagnostic method for DIVA. The residual NSP in inactivated vaccines can be detected using 9E2-HRP, which indicated the MAb 9E2 is able to evaluate inactivated vaccines. The four–amino acid epitope is the first reported to date that is recognized by 1E10. These results provide valuable insight into the diagnosis of DIVA and the NSP residual evaluation in inactivated vaccines.
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References
Bi C, Shao Z, Li J, Weng C (2019) Identification of novel epitopes targeting non-structural protein 2 of PRRSV using monoclonal antibodies. Appl Microbiol Biotechnol 103(6):2689–2699
Biswal JK, Jena S, Mohapatra JK, Bisht P, Pattnaik B (2014) Detection of antibodies specific for foot-and-mouth disease virus infection using indirect ELISA based on recombinant nonstructural protein 2B. Arch Virol 159(7):1641–1650
Brocchi E, Bergmann IE, Dekker A, Paton DJ, Sammin DJ, Greiner M, Grazioli S, Simone FD, Yadin H, Haas B (2006) Comparative evaluation of six ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus. Vaccine 24(47):6966–6979
Chung CJ, Clavijo A, Bounpheng MA, Uddowla S, Sayed A, Dancho B, Olesen IC, Pacheco J, Kamicker BJ, Brake DA, Bandaranayaka-Mudiyanselage CL, Lee SS, Rai DK, Rieder E (2018) An improved, rapid competitive ELISA using a novel conserved 3B epitope for the detection of serum antibodies to foot-and-mouth disease virus. J Vet Diagn Invest 30(5):699–707
Dreumel AKV, Michalski WP, Mcnabb LM, Shiell BJ, Singanallur NB, Peck GR (2015) Pan-serotype diagnostic for foot-and-mouth disease using the consensus antigen of nonstructural protein 3B. J Clin Microbiol 53(6):1797–1805
Fan L, Jong MH, Yang DW (2006) Presence of antibodies to non-structural proteins of foot-and-mouth disease virus in repeatedly vaccinated cattle. Vet Microbiol 115(1):14–20
Fu Y, Lu Z, Li P, Cao Y, Sun P, Tian M, Wang N, Bao H, Bai X, Li D (2014) Development of a blocking ELISA based on a monoclonal antibody against a predominant epitope in non-structural protein 3B2 of foot-and-mouth disease virus for differentiating infected from vaccinated animals. PLoS One 9(11):e111737
Fu Y, Li P, Cao Y, Wang N, Sun P, Shi Q, Ji X, Bao H, Li D, Chen Y (2017) Development of a blocking ELISA using a monoclonal antibody to a dominant epitope in non-structural protein 3A of foot-and-mouth disease virus, as a matching test for a negative-marker vaccine. PLoS One 12(1):e0170560
Gao M, Zhang R, Li M, Li S, Cao Y, Ma B, Wang J (2012) An ELISA based on the repeated foot-and-mouth disease virus 3B epitope peptide can distinguish infected and vaccinated cattle. Appl Microbiol Biotechnol 93(3):1271–1279
Gao Y, Sun SQ, Guo HC (2016) Biological function of foot-and-mouth disease virus non-structural proteins and non-coding elements. Virol J 13(1):107
Höhlich BJ, Wiesmüller KH, Schlapp T, Haas B, Pfaff E, Saalmüller A (2003a) Identification of foot-and-mouth disease virus-specific linear B-cell epitopes to differentiate between infected and vaccinated cattle. J Virol 77(16):8633–8639
Höhlich BJ, Wiesmüller KH, Haas B, Gerner W, Correa R, Hehnen HR, Schlapp T, Pfaff E, Saalmüller A (2003b) Induction of an antigen-specific immune response and partial protection of cattle against challenge infection with foot-and-mouth disease virus (FMDV) after lipopeptide vaccination with FMDV-specific B-cell epitopes. J Gen Virol 84(Pt 12):3315–3324
Hosamani M, Basagoudanavar SH, Tamil Selvan RP, Das V, Ngangom P, Sreenivasa BP, Hegde R, Venkataramanan R (2015) A multi-species indirect ELISA for detection of non-structural protein 3ABC specific antibodies to foot-and-mouth disease virus. Arch Virol 160(4):937–944
Inoue T, Parida S, Paton DJ, Linchongsubongkoch W, Mackay D, Oh Y, Aunpomma D, Gubbins S, Saeki T (2006) Development and evaluation of an indirect enzyme-linked immunosorbent assay for detection of foot-and-mouth disease virus nonstructural protein antibody using a chemically synthesized 2B peptide as antigen. J Vet Diagn Investig 18(6):545–552
Jamal SM, Belsham GJ (2013) Foot-and-mouth disease: past, present and future. Vet Res 44(1):116
Jaworski JP, Compaired D, Trotta M, Perez M, Trono K, Fondevila N (2011) Validation of an r3AB1-FMDV-NSP ELISA to distinguish between cattle infected and vaccinated with foot-and-mouth disease virus. J Virol Methods 178(1-2):191–200
Kumar N, Sharma R, Kakker NK (2007) Non-structural protein 3A for differentiation of foot-and-mouth disease infected and vaccinated animals in Haryana (India). Zoonoses Public Health 54(9-10):376–382
Li C, Liang W, Liu W, Yang D, Wang H, Ma W, Zhou G, Yu L (2016) Identification of a conserved linear epitope using a monoclonal antibody against non-structural protein 3B of foot-and-mouth disease virus. Arch Virol 161(2):365–375
Liu W, Yang B, Wang M, Wang H, Yang D, Ma W, Zhou G, Yu L (2017a) Identification of a conformational neutralizing epitope on the VP1 protein of type A foot-and-mouth disease virus. Res Vet Sci 115:374–381
Liu Z, Shao J, Zhao F, Zhou G, Gao S, Liu W, Lv J, Li X, Li Y, Chang H (2017b) Chemiluminescence immunoassay for the detection of antibodies against the 2C and 3ABC nonstructural proteins induced by infecting pigs with the foot-and-mouth disease virus. Clin Vaccine Immunol 24(8):CVI.00153-17
Liu Z-Z, Zhao F-R, Gao S-D, Shao J-J, Zhang Y-G, Chang H-Y (2018) Development of a chemiluminescence immunoassay using recombinant non-structural epitope-based proteins to accurately differentiate foot-and-mouth disease virus-infected and vaccinated bovines. Transbound Emerg Dis 65(2):338–344
Lu ZJ, Zhang XL, Fu YF, Cao YM, Tian MN, Pu S, Dong L, Liu ZX, Xie QG (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(1-2):128–133
Mahajan S, Mohapatra JK, Pandey LK, Sharma GK, Pattnaik B (2015) Indirect ELISA using recombinant nonstructural protein 3D to detect foot and mouth disease virus infection associated antibodies. Biologicals 43(1):47–54
Meyer RF, Babcock GD, Newman JFE, 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(1):33–43
Mohapatra JK, Pandey LK, Sanyal A, Pattnaik B (2011) Recombinant non-structural polyprotein 3AB-based serodiagnostic strategy for FMD surveillance in bovines irrespective of vaccination. J Virol Methods 177(2):184–192
Mohapatra AK, Mohapatra JK, Pandey LK, Sanyal A, Pattnaik B (2014) Diagnostic potential of recombinant nonstructural protein 3B to detect antibodies induced by foot-and-mouth disease virus infection in bovines. Arch Virol 159(9):2359–2369
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(4):317–325
Pacheco JM, Gladue DP, Holinka LG, Arzt J, Bishop E, Smoliga G, Pauszek SJ, Bracht AJ, O’Donnell V, Fernandez-Sainz I (2013) A partial deletion in non-structural protein 3A can attenuate foot-and-mouth disease virus in cattle. Virology 446(1-2):260–267
Robiolo B, Seki C, Fondevilla N, Grigera P, Scodeller E, Periolo O, Torre JL, Mattion N (2006) Analysis of the immune response to FMDV structural and non-structural proteins in cattle in Argentina by the combined use of liquid phase and 3ABC-ELISA tests. Vaccine 24(7):997–1008
Shourian M, Ghourchian H, Boutorabi M (2015) Ultra-sensitive immunosensor for detection of hepatitis B surface antigen using multi-functionalized gold nanoparticles. Anal Chim Acta 895:1–11
Sørensen 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(8):1461–1476
Stenfeldt C, Arzt J, Pacheco JM, Gladue DP, Smoliga GR, Silva EB, Rodriguez LL, Borca MV (2018) A partial deletion within foot-and-mouth disease virus non-structural protein 3A causes clinical attenuation in cattle but does not prevent subclinical infection. Virology 516:115–126
Wang M, Xu Z, Liu W, Li M, Wang H, Yang D, Ma W, Zhou G, Yu L (2019) Identification of a conserved linear epitope using monoclonal antibody against non-structural protein 3A of foot-and-mouth disease virus with potential for differentiation between infected and vaccinated animals. Res Vet Sci 124:178–185
Yang PC, Chu RM, Chung WB, Sung HT (1999) Epidemiological characteristics and financial costs of the 1997 foot-and-mouth disease epidemic in Taiwan. Vet Rec 145:731–734
Yang SZ, Yang JF, Zhang GP, Qiao SL, Wang XN, Zhao D, Li XW, Deng RG, Zhi AM, You LM (2010) Development of a peptide-based immunochromatographic strip for differentiation of serotype O Foot-and-mouth disease virus-infected pigs from vaccinated pigs. J Vet Diagn Investig 22(3):412–415
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This study was supported by grants from the National Key R&D Program of China (2017YFD0500902 and 2016YFE0204100).
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Liu, W., Shao, J., Chen, D. et al. Identification of three linear B cell epitopes against non-structural protein 3ABC of FMDV using monoclonal antibodies. Appl Microbiol Biotechnol 103, 8075–8086 (2019). https://doi.org/10.1007/s00253-019-10081-0
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DOI: https://doi.org/10.1007/s00253-019-10081-0