RRC ID 57000
著者 Masuda A, Lee JM, Miyata T, Sato T, Hayashi S, Hino M, Morokuma D, Karasaki N, Mon H, Kusakabe T.
タイトル Purification and characterization of immunogenic recombinant virus-like particles of porcine circovirus type 2 expressed in silkworm pupae.
ジャーナル J Gen Virol
Abstract Porcine circovirus type 2 (PCV2) is a primary causative agent of postweaningmultisystemic wasting syndrome (PMWS), which has a significant economic impact on the swine industry. The capsid protein (Cap) encoded by ORF2 of the viral genome has been used effectively as a vaccine against PCV2 infection. The Cap protein can spontaneously assemble into virus-like particles (VLPs) that are safe and highly immunogenic for vaccine applications. Several expression systems, including bacteria, yeast and insect cells, have been utilized to produce PCV2 VLPs. However, in some cases, the recombinant Cap (rCap) proteins produced in bacteria and yeast do not assemble spontaneously. In this study, we expressed rCap protein using a silkworm-baculovirus expression vector system (silkworm-BEVS) for mass production of PCV2 VLPs and established a simple three-step protocol for its purification from pupae: extraction by detergent, ammonium sulfate precipitation and anion exchange column chromatography. Size-exclusion chromatography (SEC) analysis and transmission electron microscope (TEM) observation showed that purified rCap proteins formed VLPs with a similar morphology to that of the original virus. Furthermore, the VLPs produced in silkworms were capable of inducing neutralizing antibodies against PCV2 in mice. Our results demonstrated that the silkworm system is a powerful tool for the production of PCV2 VLPs and will be useful for the development of a reliable and cost-effective PCV2 vaccine.
巻・号 99(7)
ページ 917-926
公開日 2018-7-1
DOI 10.1099/jgv.0.001087
PMID 29851377
MeSH Animals Antibodies, Neutralizing / biosynthesis Antibodies, Viral / biosynthesis Baculoviridae / genetics Baculoviridae / metabolism Bombyx / genetics Bombyx / growth & development Bombyx / virology* Capsid Proteins / administration & dosage Capsid Proteins / genetics Capsid Proteins / immunology Capsid Proteins / isolation & purification* Circovirus / drug effects* Circovirus / genetics Circovirus / immunology Cloning, Molecular Female Gene Expression Genetic Vectors / chemistry Genetic Vectors / metabolism Immunogenicity, Vaccine Mice Mice, Inbred BALB C Porcine Postweaning Multisystemic Wasting Syndrome / immunology Porcine Postweaning Multisystemic Wasting Syndrome / prevention & control* Porcine Postweaning Multisystemic Wasting Syndrome / virology Pupa / genetics Pupa / metabolism Pupa / virology Recombinant Proteins / administration & dosage Recombinant Proteins / genetics Recombinant Proteins / immunology Recombinant Proteins / isolation & purification Swine Vaccines, Virus-Like Particle / administration & dosage Vaccines, Virus-Like Particle / biosynthesis Vaccines, Virus-Like Particle / genetics Vaccines, Virus-Like Particle / immunology* Viral Vaccines / administration & dosage Viral Vaccines / biosynthesis Viral Vaccines / genetics Viral Vaccines / immunology*
IF 2.809
引用数 6
リソース情報
カイコ d17