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Secretion of truncated recombinant rabies virus glycoprotein with preserved antigenic properties using a co-expression system in Hansenula polymorpha

  • Virology
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Abstract

Rabies virus infection remains a serious public health threat in the developing world, where cost-concerns make wide-scale public health interventions impractical. The development of novel and inexpensive ELISA diagnostic antigens is critical in early detection and prevention of complications. The transmembrane glycoprotein (G) of rabies virus (RV) contains an external domain capable of inducing the synthesis of anti-rabies, virus-neutralizing antibodies, in infected or immunized hosts. In our study, the external G domain was synthesized and fused in-frame with a polyhistidine-tag coding sequence present in the expression plasmid. Soluble truncated recombinant G was secreted in Hansenula polymorpha (H. polymorpha) using H. polymorpha-derived calnexin (HpCNE1) overproduction and found to be correctly N-glycosylated. The truncated recombinant G was purified from cell culture supernatant by Ni-agarose affinity chromatography and when compared with the full-length glycoprotein, found to be similarly immunogenic in vaccinated rabbits. These results subsequently led us to explore the potential of truncated recombinant G as a diagnostic antigen in ELISA. Our results show that the truncated recombinant G can detect antibodies directed to both whole virion and native glycoprotein. More sophisticated applications of truncated recombinant G would profit from the correctly N-glycosylated and soluble monomer.

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Authors and Affiliations

  1. Life Science and Engineering School, Shaanxi University of Science and Technology, Xi’an, 710021, P. R. China

    Weidong Qian

  2. Northwestern University, Chicago, IL, USA

    Frank Aguilar

  3. Medical Technology and Engineering School, Henan University of Science and Technology, Luoyang, 471003, P. R. China

    Ting Wang

  4. Center for Agricultural Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, P. R. China

    Bingsheng Qiu

Authors
  1. Weidong Qian
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  2. Frank Aguilar
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  3. Ting Wang
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  4. Bingsheng Qiu
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Correspondence to Bingsheng Qiu.

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Qian, W., Aguilar, F., Wang, T. et al. Secretion of truncated recombinant rabies virus glycoprotein with preserved antigenic properties using a co-expression system in Hansenula polymorpha . J Microbiol. 51, 234–240 (2013). https://doi.org/10.1007/s12275-013-2337-0

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  • DOI: https://doi.org/10.1007/s12275-013-2337-0

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