High-level expression and purification of biologically active human IL-2 using silkworm-baculovirus expression vector system

Masato Hino, Takuji Kawanami, Jian Xu, Daisuke Morokuma, Kazuma Hirata, Mami Yamashita, Noriko Karasaki, Tuneyuki Tatsuke, Hiroaki Mon, Kazuhiro Iiyama, Noriho Kamiya, Yutaka Banno, Takahiro Kusakabe, Man Lee

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Interleukin 2 (IL-2) is a pharmacologically vital cytokine secreted mainly by activated CD4+ T lymphocyte. Recombinant human IL-2 (rhIL-2) protein has already been globally applied as an immune-therapeutic reagent for various diseases. Therefore, there is great interest in developing an active form of rhIL-2 in very large amounts and with excellent purity for clinical use. In this study, we successfully mass-expressed and purified N- or C-terminal tandem tag-fused rhIL-2 in a baculovirus expression vector system (BEVS) using silkworm larvae as factories. We confirmed that the intrinsic instability of hIL-2 causes the loss and low recovery of N-tagged rhIL-2 and that C-tagged rhIL-2 is more suitable for mass production. Furthermore, the activity of purified rhIL-2s was further validated by a cell proliferation assay of a human natural killer cell line, and both rhIL-2 proteins produced in the silkworm-BEVS exhibited comparable activity to that of the commercial E.coli-derived rhIL-2. Taken together, our results and strategies could contribute greatly to the mass production of active rhIL-2.

Original languageEnglish
Pages (from-to)313-317
Number of pages5
JournalJournal of Asia-Pacific Entomology
Volume19
Issue number2
DOIs
Publication statusPublished - Jun 1 2016

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Baculoviridae
silkworms
interleukin-2
natural killer cells
factories
purity
insect larvae
cell proliferation
cytokines
proteins
T-lymphocytes
cell lines
Escherichia coli
therapeutics
assays

All Science Journal Classification (ASJC) codes

  • Insect Science

Cite this

High-level expression and purification of biologically active human IL-2 using silkworm-baculovirus expression vector system. / Hino, Masato; Kawanami, Takuji; Xu, Jian; Morokuma, Daisuke; Hirata, Kazuma; Yamashita, Mami; Karasaki, Noriko; Tatsuke, Tuneyuki; Mon, Hiroaki; Iiyama, Kazuhiro; Kamiya, Noriho; Banno, Yutaka; Kusakabe, Takahiro; Lee, Man.

In: Journal of Asia-Pacific Entomology, Vol. 19, No. 2, 01.06.2016, p. 313-317.

Research output: Contribution to journalArticle

Hino, Masato ; Kawanami, Takuji ; Xu, Jian ; Morokuma, Daisuke ; Hirata, Kazuma ; Yamashita, Mami ; Karasaki, Noriko ; Tatsuke, Tuneyuki ; Mon, Hiroaki ; Iiyama, Kazuhiro ; Kamiya, Noriho ; Banno, Yutaka ; Kusakabe, Takahiro ; Lee, Man. / High-level expression and purification of biologically active human IL-2 using silkworm-baculovirus expression vector system. In: Journal of Asia-Pacific Entomology. 2016 ; Vol. 19, No. 2. pp. 313-317.
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