Efficient production of recombinant SARS-CoV-2 spike protein using the baculovirus-silkworm system
| العنوان: | Efficient production of recombinant SARS-CoV-2 spike protein using the baculovirus-silkworm system |
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| المؤلفون: | Ryosuke Fujita, Takato Moriyama, Yoshino Tonooka, Miyu Tanaka, Tsuguru Fujii, Takeru Ebihara, Masato Hino, Akitsu Masuda, Jae Man Lee, Hiroaki Mon, Takumi Nagasato, Takahiro Kusakabe, Ryo Nagai, Kohei Kakino |
| المصدر: | Biochemical and Biophysical Research Communications |
| بيانات النشر: | Elsevier BV, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, viruses, Biophysics, Spike protein, Biochemistry, Article, Cell Line, law.invention, 03 medical and health sciences, 0302 clinical medicine, Antigen, law, Animals, Cloning, Molecular, Silkworm-BmNPV expression system, Molecular Biology, Furin, biology, SARS-CoV-2, Chemistry, fungi, Cell Biology, Transfection, Bombyx, Molecular biology, Nucleopolyhedroviruses, Recombinant Proteins, 030104 developmental biology, Ectodomain, Immunization, 030220 oncology & carcinogenesis, Spike Glycoprotein, Coronavirus, biology.protein, Recombinant DNA, Antibody, Fetal bovine serum |
| الوصف: | 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. Highlights • Establishment of the BmNPV-silkworm expression system for production of the ectodomain of SARS-CoV-2 spike protein. • The SARS-CoV-2 S protein with the native sequence was cleaved by the host silkworm furin protease. • S protein without furin-cleavage site could be purified as oligomeric structures with immunoreactivity to the anti-SARS-CoV-2 S2 antibody. |
| تدمد: | 0006-291X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fab7b18f2d5ea12f21a6204c73ce360e https://doi.org/10.1016/j.bbrc.2020.06.020 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....fab7b18f2d5ea12f21a6204c73ce360e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 0006291X |
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