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Efficient application of a baculovirus-silkworm larvae expression system for obtaining porcine circovirus type 2 virus-like particles for a vaccine

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Abstract

Porcine circovirus type 2 (PCV2) is a major pathogen associated with swine diseases. It is the smallest single-stranded DNA virus, and its genome contains four major open reading frames (ORFs). ORF2 encodes the major structural protein Cap, which can self-assemble into virus-like particles (VLPs) in vitro and contains the primary antigenic determinants. In this study, we developed a high-efficiency method for obtaining VLPs and optimized the purification conditions. In this method, we expressed the protein Cap with a 6× His tag using baculovirus-infected silkworm larvae as well as the E. coli BL21(DE3) prokaryotic expression system. The PCV2 Cap proteins produced by the silkworm larvae and E. coli BL21(DE3) were purified. Cap proteins purified from silkworm larvae self-assembled into VLPs in vitro, while the Cap proteins purified from bacteria were unable to self-assemble. Transmission electron microscopy confirmed the self-assembly of VLPs. The immunogenicity of the VLPs produced using the baculovirus system was demonstrated using an enzyme-linked immunosorbent assay (ELISA). Furthermore, the purification process was optimized. The results demonstrated that the expression system using baculovirus-infected silkworm larvae is a good choice for obtaining VLPs of PCV2 and has potential for the development of a low-cost and efficient vaccine.

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Acknowledgements

The authors acknowledge the assistance of Bo Ning in the maintenance and collection of field samples for this work. The assistance of Dr. Feifei Ren in experimental technology is also greatly appreciated.

Funding

This work was supported by the National Natural Science Foundation of China (grant numbers 31872426 and, 31372373), the Natural Science Foundation of Guangdong Province, China (grant number 2016A030311018).

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  1. Qianhua He and Zhenming Cao contributed equally to this work.

Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Agro-animal Genomics and Molecular Breeding and Subtropical Sericulture and Mulberry Resources Protection and Safety Engineering Research Center, College of Animal Science, South China Agricultural University, Guangzhou, 510642, Guangdong, China

    Qianhua He, Zhenming Cao, Pengwei Wang, Qiuyuan Lu, Hao Zheng & **gchen Sun

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  1. Qianhua He
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  2. Zhenming Cao
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  3. Pengwei Wang
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Contributions

JS coordinated the project. QH and ZC performed the research. QH HZ and JS wrote the manuscript. QL and PW contributed to new methods. QH, ZC and JS performed data analysis. QH, ZC, and JS interpreted the context of results. All of the authors have read and approved the manuscript.

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Correspondence to Hao Zheng or **gchen Sun.

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He, Q., Cao, Z., Wang, P. et al. Efficient application of a baculovirus-silkworm larvae expression system for obtaining porcine circovirus type 2 virus-like particles for a vaccine. Arch Virol 165, 2301–2309 (2020). https://doi.org/10.1007/s00705-020-04754-9

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