Vet Med - Czech, 2018, 63(10):468-475 | DOI: 10.17221/23/2018-VETMED

Preparation and characterisation of monoclonal antibodies against the N protein of the SHpd/2012 strain of porcine epidemic diarrhoea virusOriginal Paper

S.J. Ding1, Y.R. Luo1, S.T. Zhou1, C. Xie1, K. Wang2, Y.Y. Xie1, L. Cui1, X.G. Hua1, C.L. Yuan1, Y.J. Zhou*,2, Z.B. Yang*,1
1 Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China
2 Division of Swine Infectious Diseases, Shanghai Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Shanghai, China

Porcine epidemic diarrhoea is caused by the porcine epidemic diarrhoea virus, and is a highly contagious disease which affects the intestines of new-born piglets resulting in intense diarrhoea. Historically, the virus has caused enormous economic losses in the pig industry. In particular, the emergence of new epidemic strains means there is a pressing need for prevention and control of the disease. Owing to the specificity of the monoclonal antibodies now available, study of the pathogenesis, immune mechanisms and new diagnostic methods can be performed. In this study, 13 strains of positive hybridoma cells were prepared by immunising mice with purified whole porcine epidemic diarrhoea virus, and analysis was performed using ELISA and Western blotting. Three cell strains specifically recognised the porcine epidemic diarrhoea virus nucleocapsid protein (N protein). In this study, we report the characterisation of effective tools for the establishment of porcine epidemic diarrhoea virus diagnostic methods and we have specifically generated primary antibodies for ELISA, IFA, test strip and Western blotting.

Keywords: porcine epidemic diarrhoea virus; PEDV; nucleocapsid protein; N protein; monoclonal antibodies; MAbs

Published: October 31, 2018  Show citation

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Ding SJ, Luo YR, Zhou ST, Xie C, Wang K, Xie YY, et al.. Preparation and characterisation of monoclonal antibodies against the N protein of the SHpd/2012 strain of porcine epidemic diarrhoea virus. Vet Med - Czech. 2018;63(10):468-475. doi: 10.17221/23/2018-VETMED.
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    References

    1. Chen F, Zhu Y, Wu M, Ku X, Ye S, Li Z, Guo X, He Q (2015): Comparative genomic analysis of classical and variant virulent parental/attenuated strains of porcine epidemic diarrhoea virus. Viruses 7, 5525-5538. Go to original source... Go to PubMed...
    2. Cheng Q, Niu X, Ye C (1992): Investigation of porcine epidemic diarrhoea disease in Qinghai area. Journal of Animal Science and Veterinary Medicine in Qinghai 22, 22-23.
    3. Feng L, Sun DB, Chen JF, Shi HY, Wu BP, Cui XC, Lv MJ, Liu D, Tong GZ (2008): Preparation and characterization of the monoclonal antibodies against nucleocapsid protein of PEDV. Chinese Journal of Preventive Veterinary Medicine 30, 190-193.
    4. Gao Y, Kou Q, Ge X, Zhou L, Guo X, Yang H (2013): Phylogenetic analysis of porcine epidemic diarrhoea virus field strains prevailing recently in China. Archives of Virology 158, 711-715. Go to original source... Go to PubMed...
    5. Lee HK, Yeo SG (2003): Cloning and sequence analysis of the nucleocapsid gene of porcine epidemic diarrhoea virus Chinju99. Virus Genes 26, 207-212. Go to original source... Go to PubMed...
    6. Lei X, Zhao J, Wang X, Zhao Y, Wang C (2015): Development of a hybridoma cell line secreting monoclonal antibody against S protein of a Chinese variant of PEDV. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy 34, 12-16. Go to original source... Go to PubMed...
    7. Lei XM, Chen J, Yang PP, Zhao YX, Wang XM, Zhao J, Wang CQ (2016): The development of the hybridoma cell line secreting monoclonal antibody against S protein of a PEDV variant. Chinese Journal of Veterinary Science 36, 206-215.
    8. Li ZL, Zhu L, Ma JY, Zhou QF, Song YH, Sun BL, Chen RA, Xie QM, Bee YZ (2012a): Molecular characterization and phylogenetic analysis of porcine epidemic diarrhoea virus (PEDV) field strains in south China. Virus Genes 45, 181- 185. Go to original source... Go to PubMed...
    9. Li W, Li H, Liu Y, Pan Y, Deng F, Song Y, Tang X, He Q (2012b): New variants of porcine epidemic diarrhoea virus, China, 2011. Emerging Infectious Diseases 18, 1350-1353. Go to original source...
    10. Li R, Qiao S, Yang Y, Su Y, Zhao P, Zhou E, Zhang G (2014): Phylogenetic analysis of porcine epidemic diarrhoea virus (PEDV) field strains in central China based on the ORF3 gene and the main neutralization epitopes. Archives of Virology 159, 1057-1065. Go to original source... Go to PubMed...
    11. Li R, Qiao S, Yang Y, Guo J, Xie S, Zhou E, Zhang G (2016): Genome sequencing and analysis of a novel recombinant porcine epidemic diarrhoea virus strain from Henan, China. Virus Genes 52, 91-98. Go to original source... Go to PubMed...
    12. Liu SJ, Duan SH, Hu JY, Jin JJ, Tu YQ, Bai Y (2015): Preparation and identification of monoclonal antibodies against the nucleocapsid protein of porcine epidemic diarrhoea virus. Acta Agriculturae Zhejiangensis 27, 950-954.
    13. Oldham J (1972): Pig Farming (Oct suppl), 72-73.
    14. Pei DG, Ge JW, Ma GP, Jiang YP, Li YJ (2008): Production and identification of monoclonal antibodies against recombinant PEDV N protein. Journal of Northeast Agriculture University 39, 84-89.
    15. Rodak L, Valicek L, Smid B, Nevorankova Z (2005): An ELISA optimized for porcine epidemic diarrhoea virus detection in faeces. Veterinary Microbiology 105, 9-17. Go to original source... Go to PubMed...
    16. Shi XL, Tan C, Zhao PD, Bai J, Li YF, Jiang P (2014): Preparation and identification of monoclonal antibodies against the nucleocapsid protein of porcine epidemic diarrhoea virus. Animal husbandry and veterinary medicine 46, 1-5.
    17. Song D, Park B (2012): Porcine epidemic diarrhoea virus: a comprehensive review of molecular epidemiology, diagnosis, and vaccines. Virus Genes 44, 167-175. Go to original source... Go to PubMed...
    18. Sun DB, Feng L, Shi HY, Chen JF, Cui XC, Tong YE (2007): Preparation and characterization of the monoclonal antibodies against neutralizing epitope region of PEDV S protein. Chinese Journal of Preventive Veterinary Medicine 29, 887-890.
    19. Sun RQ, Cai RJ, Chen YQ, Liang PS, Chen DK, Song CX (2012): Outbreak of porcine epidemic diarrhoea in suckling piglets, China. Emerging Infectious Diseases 18, 161-163. Go to original source... Go to PubMed...
    20. Sun D, Wang X, Wei S, Chen J, Feng L (2016): Epidemiology and Vaccine of porcine epidemic diarrhoea virus in China: mini-review. Journal of Veterinary Medical Science 78, 355-363. Go to original source... Go to PubMed...
    21. Wang J, Zhao P, Guo L, Liu Y, Du Y, Ren S, Li J, Zhang Y, Fan Y, Huang B, Liu S, Wu J (2013): Porcine epidemic diarrhoea virus variants with high pathogenicity, China. Emerging Infectious Diseases 19, 2048-2049. Go to original source... Go to PubMed...
    22. Wang Z, Jiyuan Y, Su C, Xinyuan Q, Lijie T, Yijing L (2015a): Development of an antigen capture enzyme-linked immunosorbent assay for virus detection based on porcine epidemic diarrhoea virus monoclonal antibodies. Viral Immunology 28, 184-189. Go to original source... Go to PubMed...
    23. Wang J, Chen J, Wei F, Dong Y, Zhu L, Han W, Wang L, Shen Z (2015b): prokaryotic expression of truncated S1 protein of porcine epidemic diarrhoea virus and production of monoclonal antibodies to recombinant protein. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy 34, 327-333. Go to original source... Go to PubMed...
    24. Wang K, Xie C, Zhang J, Zhang W, Yang D, Yu L, Jiang Y, Yang S, Gao F, Yang Z, Zhou Y, Tong G (2016): The identification and characterization of two novel epitopes on the nucleocapsid protein of the porcine epidemic diarrhoea virus. Scientific Reports 6, doi: 10.1038/srep39010. Go to original source... Go to PubMed...
    25. Wang MY, Lu HB, Liu YJ, Guo CM, Zhu LS, Xing ZL, Deng YR, Ouyang HS, Wang XP (2017): Generation of monoclonal antibody against PEDV S protein and detection of PEDV antigens using the developed sandwich ELISA. Chinese Journal of Veterinary Science 37, 2033-2042.
    26. Wu Y, Yu L, Cheng Q, Wang K, Yu H, Liu G, Tong G, Zhou Y (2012): Expression and identification of M gene of porcine epidemic diarrhoea virus. Chinese Journal of Animal Infectious Diseases 20, 6-10.
    27. Zhang ZB, Chen JF, Shi HY, Yang B, Shi D, Chen XJ, Feng L (2011): Preparation of the monoclonal antibody against membrane protein of porcine epidemic diarrhoea virus. Chinese Journal of Preventive Veterinary Medicine 33, 568-570.
    28. Zhang Q, Hu R, Tang X, Wu C, He Q, Zhao Z, Chen H, Wu B (2013): Occurrence and investigation of enteric viral infections in pigs with diarrhoea in China. Archives of Virology 158, 1631-1636. Go to original source... Go to PubMed...
    29. Zheng FM, Zhao J, Huo JY, Li Huan, Yang X, Chen L, Chang HT, Wang XW, Liu HY, Du JM, Yao HX,Wang CQ (2014): Prokaryotic expression of N gene major antigen region of PEDV and development of an indirect ELISA based on expressed protein. Chinese Journal of Veterinary Science 34, 371-378.

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