Production of a biologically active human basic fibroblast growth factor using silkworm-baculovirus expression vector system

Akitsu Masuda, Jian Xu, Kosuke Minamihata, Genki Kagawa, Yusei Hamada, Yoshiki Morifuji, Takumi Yano, Masato Hino, Daisuke Morokuma, Noriko Karasaki, Hiroaki Mon, Noriho Kamiya, Takahiro Kusakabe, Man Lee

研究成果: ジャーナルへの寄稿記事

抄録

As a therapeutic treatment, recombinant human basic fibroblast growth factor (rhbFGF) is usually employed in tissue regeneration, and as an essential component in culture medium for maintaining the induced pluripotent stem (iPS) cell and embryonic stem (ES) cell in an undifferentiated state. Therefore, a large amount of biologically active rhbFGF is required. In this study, silkworm-baculovirus expression vector system (silkworm-BEVS) is employed to achieve a high productivity of recombinant rhbFGF with two small affinity tags (His-tag and STREP-tag) at the N or C-terminus. It is observed that rhbFGF with 30 K signal peptide of silkworm were successfully expressed but are not sufficiently secreted into the culture medium of cultured insect cells. Then we purified the N- or C-tagged intracellular rhbFGF protein and obtained a yield of about 0.7 mg/larva and 1.2 mg/larva, respectively. Although the final yield of the C-tagged rhbFGF is higher than that of the N-tagged, rhbFGF with N-tag demonstrated promising and comparable biological activity, which is evaluated through a mammalian cell proliferation assay. Taken together, these results indicate that silkworm-BEVS could contribute to the mass-production of the biologically active rhbFGF for medical uses.

元の言語英語
ページ(範囲)716-720
ページ数5
ジャーナルJournal of Asia-Pacific Entomology
21
発行部数2
DOI
出版物ステータス出版済み - 6 1 2018

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fibroblast growth factor 2
Baculoviridae
silkworms
culture media
larvae
embryonic stem cells
signal peptide
tissue repair
bioactive properties
cell proliferation
therapeutics
insects
assays

All Science Journal Classification (ASJC) codes

  • Insect Science

これを引用

Production of a biologically active human basic fibroblast growth factor using silkworm-baculovirus expression vector system. / Masuda, Akitsu; Xu, Jian; Minamihata, Kosuke; Kagawa, Genki; Hamada, Yusei; Morifuji, Yoshiki; Yano, Takumi; Hino, Masato; Morokuma, Daisuke; Karasaki, Noriko; Mon, Hiroaki; Kamiya, Noriho; Kusakabe, Takahiro; Lee, Man.

:: Journal of Asia-Pacific Entomology, 巻 21, 番号 2, 01.06.2018, p. 716-720.

研究成果: ジャーナルへの寄稿記事

Masuda, A, Xu, J, Minamihata, K, Kagawa, G, Hamada, Y, Morifuji, Y, Yano, T, Hino, M, Morokuma, D, Karasaki, N, Mon, H, Kamiya, N, Kusakabe, T & Lee, M 2018, 'Production of a biologically active human basic fibroblast growth factor using silkworm-baculovirus expression vector system', Journal of Asia-Pacific Entomology, 巻. 21, 番号 2, pp. 716-720. https://doi.org/10.1016/j.aspen.2018.05.002
Masuda, Akitsu ; Xu, Jian ; Minamihata, Kosuke ; Kagawa, Genki ; Hamada, Yusei ; Morifuji, Yoshiki ; Yano, Takumi ; Hino, Masato ; Morokuma, Daisuke ; Karasaki, Noriko ; Mon, Hiroaki ; Kamiya, Noriho ; Kusakabe, Takahiro ; Lee, Man. / Production of a biologically active human basic fibroblast growth factor using silkworm-baculovirus expression vector system. :: Journal of Asia-Pacific Entomology. 2018 ; 巻 21, 番号 2. pp. 716-720.
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AU - Masuda, Akitsu

AU - Xu, Jian

AU - Minamihata, Kosuke

AU - Kagawa, Genki

AU - Hamada, Yusei

AU - Morifuji, Yoshiki

AU - Yano, Takumi

AU - Hino, Masato

AU - Morokuma, Daisuke

AU - Karasaki, Noriko

AU - Mon, Hiroaki

AU - Kamiya, Noriho

AU - Kusakabe, Takahiro

AU - Lee, Man

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