Applied Microbiology and Biotechnology

, Volume 103, Issue 19, pp 8075–8086 | Cite as

Identification of three linear B cell epitopes against non-structural protein 3ABC of FMDV using monoclonal antibodies

  • Wei Liu
  • Junjun Shao
  • Danian Chen
  • Yanyan Chang
  • Huiyun ChangEmail author
  • Yongguang ZhangEmail author
Applied genetics and molecular biotechnology


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.


Foot-and-mouth disease virus Non-structural protein 3ABC Monoclonal antibodies Epitope mapping 


Funding information

This study was supported by grants from the National Key R&D Program of China (2017YFD0500902 and 2016YFE0204100).

Compliance with ethical standards

All procedures involving animals were approved by the Animal Ethics Committee of LVRI, Chinese Academy of Agricultural Sciences.

Conflict of interest

The authors declare that they have no conflicting interests.

Ethics statement

All applicable international, national, and institutional guidelines for the care and use of animals were followed.

Supplementary material

253_2019_10081_MOESM1_ESM.pdf (332 kb)
ESM 1 (PDF 331 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research InstituteChinese Academy of Agricultural SciencesLanzhouChina
  2. 2.Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhouChina

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