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Characterisation of novel linear antigen epitopes on North American-type porcine reproductive and respiratory syndrome virus M protein

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Abstract

The M protein, encoded by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF6 gene, is considered to be one of the most conserved PRRSV proteins. In recent decades, highly specific monoclonal antibodies (Mabs) have been exploited to provide reliable diagnoses for many diseases. In this study, two different Mab clones targeting the linear epitopes on the PRRSV M protein were generated and characterized. Both Mabs showed binding activity against the native PRRSV virion and recombinant M protein when analyzed by immunofluorescence assay (IFA) and Western blot. The targeted epitope of each Mab was mapped by serial truncation of the M protein to generate overlapping fragments. Fine epitope mapping was then performed using a panel of expressed polypeptides. The polypeptide sequences of the two epitopes recognized by Mabs 1C8 and 3F7 were 3SSLD6 and 155VLGGRKAVK163, respectively, with the former being a newly identified epitope on the M protein. In both cases, these two epitopes were finely mapped for the first time. Alignments of Mab epitope sequences revealed that the two epitopes on the M protein were highly conserved between the North American-type strains. These Mabs, along with their mapped epitopes, are useful for the development of diagnostic and research tools, including immunofluorescence, ELISA and Western blot.

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Acknowledgments

This study was supported by the Heilongjiang Natural Science Funds for Distinguished Young Scholars (JC201314), the National Natural Science Foundation of China (grant no. 31001050) and the National High Technology Research and Development Program (863 plan) (Grant No. 2011AA10A208).

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Authors and Affiliations

  1. Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine and State Key Laboratory of Agribiotechnology, China Agricultural University, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100193, China

    Qian Wang, Xin Guo, Xinna Ge & Hanchun Yang

  2. Division of Swine Infectious Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 427, Maduan Street, Nangang District, Harbin, 150001, China

    Qian Wang, Jiazeng Chen, Jinmei Peng, Tongqing An, Chaoliang Leng, Yan Sun & Zhijun Tian

Authors
  1. Qian Wang
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  2. Jiazeng Chen
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  3. Jinmei Peng
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  4. Tongqing An
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  5. Chaoliang Leng
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  6. Yan Sun
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  7. Xin Guo
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  8. Xinna Ge
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  9. Zhijun Tian
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  10. Hanchun Yang
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Correspondence to Zhijun Tian or Hanchun Yang.

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Wang, Q., Chen, J., Peng, J. et al. Characterisation of novel linear antigen epitopes on North American-type porcine reproductive and respiratory syndrome virus M protein. Arch Virol 159, 3021–3028 (2014). https://doi.org/10.1007/s00705-014-2174-4

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  • DOI: https://doi.org/10.1007/s00705-014-2174-4

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