| RRC ID |
64955
|
| 著者 |
Fujita R, Hino M, Ebihara T, Nagasato T, Masuda A, Lee JM, Fujii T, Mon H, Kakino K, Nagai R, Tanaka M, Tonooka Y, Moriyama T, Kusakabe T.
|
| タイトル |
Efficient production of recombinant SARS-CoV-2 spike protein using the baculovirus-silkworm system.
|
| ジャーナル |
Biochem Biophys Res Commun
|
| Abstract |
In the case of a new viral disease outbreak, an immediate development of virus detection kits and vaccines is required. For COVID-19, we established a rapid production procedure for SARS-CoV-2 spike protein (S protein) by using the baculovirus-silkworm expression system. The baculovirus vector-derived S proteins were successfully secreted to silkworm serum, whereas those formed insoluble structure in the larval fat body and the pupal cells. The ectodomain of S protein with the native sequence was cleaved by the host furin-protease, resulting in less recombinant protein production. The S protein modified in furin protease-target site was efficiently secreted to silkworm serum and was purified as oligomers, which showed immunoreactivity for anti-SARS-CoV-2 S2 antibody. By using the direct transfection of recombinant bacmid to silkworms, we achieved the efficient production of SARS-CoV-2 S protein as fetal bovine serum (FBS)-free system. The resultant purified S protein would be useful tools for the development of immunodetection kits, antigen for immunization for immunoglobulin production, and vaccines.
|
| 巻・号 |
529(2)
|
| ページ |
257-262
|
| 公開日 |
2020-8-20
|
| DOI |
10.1016/j.bbrc.2020.06.020
|
| PII |
S0006-291X(20)31214-6
|
| PMID |
32703420
|
| PMC |
PMC7280120
|
| MeSH |
Animals
Bombyx / cytology*
Bombyx / enzymology
Bombyx / virology*
Cell Line
Cloning, Molecular
Furin / metabolism
Nucleopolyhedroviruses / genetics*
Nucleopolyhedroviruses / metabolism
Recombinant Proteins / biosynthesis
Recombinant Proteins / genetics
Spike Glycoprotein, Coronavirus / biosynthesis*
Spike Glycoprotein, Coronavirus / chemistry
Spike Glycoprotein, Coronavirus / genetics
Spike Glycoprotein, Coronavirus / isolation & purification*
|
| IF |
2.985
|
| リソース情報 |
| カイコ |
d17
f38 |
