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Soluble expression and purification of Brucella cell surface protein (BCSP31) of Brucella melitensis and preparation of anti-BCSP31 monoclonal antibodies

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Abstract

Brucella cell surface protein (BCSP31) is potentially useful for diagnosing brucellosis. We aimed to establish a monoclonal antibody (MAb) against Brucella melitensis BCSP31 and to investigate its distribution in diagnosis. Soluble recombinant BCSP31 was successfully expressed and purified. Two MAbs (1F1 and 1E5) against B. melitensis BCSP31, effective in detecting both recombinant and cellular proteins, were obtained and characterized. The MAbs did not react with Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Mycobacterium tuberculosis, or Bacillus aeruginosus, but strongly reacted with BCSP31 and B. melitensis by ELISA and Western blot analysis. We also tested different Brucella species and brucellosis using the prepared anti-BCSP31 MAbs. BCSP31 and anti-BCSP31 MAbs may play important roles in future research in diagnosing brucellosis.

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Acknowledgments

We thank Professor Wenyu Ma, and Dr. Jun** Ren for their advice and assistance. Ailin Fan (**jng Hospital, **’an, China) and Jie Zhang (Center of Experiments, The Fourth Military Medical University) generously supplied the bacterial strains. This work was supported by grants from the Military Medical Science and Technique Foundation (No. 06G091) and National Natural Science Foundation of China (No. 30901281).

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Authors and Affiliations

  1. Department of Microbiology, and PLA Medical Key Library of Molecular Virology and Immunology, Fourth Military Medical University, **’an, 710032, People’s Republic of China

    Lei Zhang, **ng An Wu, Fang Lin Zhang, Wen Tao Bai & Zhi Kai Xu

  2. Department of Pests and Brucellosis, Shaanxi Center for Disease Control and Prevention, **’an, People’s Republic of China

    Cui Hong An & Yang **n Sun

Authors
  1. Lei Zhang
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  2. **ng An Wu
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  3. Fang Lin Zhang
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  4. Cui Hong An
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  5. Yang **n Sun
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  6. Wen Tao Bai
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  7. Zhi Kai Xu
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Corresponding authors

Correspondence to Wen Tao Bai or Zhi Kai Xu.

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Lei Zhang and **ng An Wu contributed equally to this work.

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Zhang, L., Wu, X.A., Zhang, F.L. et al. Soluble expression and purification of Brucella cell surface protein (BCSP31) of Brucella melitensis and preparation of anti-BCSP31 monoclonal antibodies. Mol Biol Rep 39, 431–438 (2012). https://doi.org/10.1007/s11033-011-0755-9

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