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

• Published in: Journal of Asia-Pacific entomology Vol. 21; no. 2; pp. 716 - 720
• Main Authors: 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, Jae Man
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01-06-2018; 한국응용곤충학회
• Subjects: Basic fibroblast growth factor; Signal peptide; Silkworm-baculovirus expression vector system; 농수해양학; Signal peptide; Basic fibroblast growth factor; Silkworm-baculovirus expression vector system
• ISSN: 1226-8615; 1876-7990
• DOI: 10.1016/j.aspen.2018.05.002

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.
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•Recombinant hbFGF were expressed in silkworm-BEVS.•The N-terminal tagged rhbFGF exhibited comparable cell proliferation activity to that of the commercial E.coli-derived rhbFGF.•The N-terminal tagged rhbFGF is more suitable for mass production in silkworm-BEVS.