MS2 Phage-mediated Preparation and Immunogenicity of Chimeric Nanoparticles with Linear Epitope of Reproductive and Respiratory Syndrome Virus

WANG Guoqiang; LI Xinxin; SU Yunfang; ZENG Huahui; MA Hongfang; LIU Baoguang; SHANG Lizhi; ZHANG Zhenqiang

doi:10.19303/j.issn.1008-0384.2020.06.007

Volume 35 Issue 6

Jun. 2020

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Article Navigation > Fujian Journal of Agricultural Sciences > 2020 > 35(6): 618-625

WANG G Q, LI X X, SU Y F, et al. MS2 Phage-mediated Preparation and Immunogenicity of Chimeric Nanoparticles with Linear Epitope of Reproductive and Respiratory Syndrome Virus [J]. Fujian Journal of Agricultural Sciences，2020，35（6）：618−625 doi: 10.19303/j.issn.1008-0384.2020.06.007

Citation:

WANG G Q, LI X X, SU Y F, et al. MS2 Phage-mediated Preparation and Immunogenicity of Chimeric Nanoparticles with Linear Epitope of Reproductive and Respiratory Syndrome Virus [J]. Fujian Journal of Agricultural Sciences，2020，35（6）：618−625 doi: 10.19303/j.issn.1008-0384.2020.06.007

Citation:

WANG G Q, LI X X, SU Y F, et al. MS2 Phage-mediated Preparation and Immunogenicity of Chimeric Nanoparticles with Linear Epitope of Reproductive and Respiratory Syndrome Virus [J]. Fujian Journal of Agricultural Sciences，2020，35（6）：618−625 doi: 10.19303/j.issn.1008-0384.2020.06.007

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MS2 Phage-mediated Preparation and Immunogenicity of Chimeric Nanoparticles with Linear Epitope of Reproductive and Respiratory Syndrome Virus

doi: 10.19303/j.issn.1008-0384.2020.06.007

1.
Academy of Traditional Chinese Medicine, Henan University of Chinese Medicine, Zhengzhou, Henan　450046, China
2.
The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, Henan　450046, China
3.
College of Animal and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan　450002, China

Received Date: 2019-12-24
Rev Recd Date: 2020-03-19
Publish Date: 2020-08-10

Abstract

Abstract

Objective Using the protein on coat of MS2 bacteriophage as a carrier, chimeric nanoparticles with a linear epitope of porcine reproductive and respiratory syndrome virus, GP5, were constructed, and their immunogenicity determined. Method The dominant neutralizing epitope gene sequence on GP5 was inserted into the MS2 protein by overlapping extension PCR. The recombinant vector was to express the chimeric protein through a prokaryotic expression system. The target protein was purified by ammonium sulfate precipitation and gel filtration chromatography followed by characterization with dynamic light scattering and electron microscopy. The immunogenicity of the constructed chimeric nanoparticles was determined by Western blot and animal immunoassay. Result The linear epitope was successfully inserted into the target sequence, and the recombinant vector reached a purity greater than 85%. The purified target protein self-assembled in vitro to form uniform chimeric nanoparticles with a diameter of 25–31 nm. After immunizing animals, the chimeric particles produced a high-level of antibodies that reacted with inactivated viruses showing significant immunogenicity. Conclusion The MS2 phage coat protein allowed the insertion of 9 exogenous polypeptides (the linear epitope on GP5) and self-assembled in vitro to form the chimeric virus-like particles. Each particle carried the exogenous polypeptides on the surface generating high immunogenicity. The current technology provided a new venue for building other epitopes of porcine reproductive and respiratory syndrome virus or epitopes with longer tandem.
- MS2 phage,
- linear epitope,
- chimeric nanoparticle,
- porcine reproductive and respiratory syndrome virus

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References(28)

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