Optimal silkworm larva host for high-level production of Mus musculus IL-4 using a baculovirus expression vector system

Masahiko Kobayashi; Jian Xu; Kohei Kakino; Akitsu Masuda; Masato Hino; Naoki Fujimoto; Kosuke Minamihata; Noriho Kamiya; Hiroaki Mon; Hiroshi Iida; Masateru Takahashi; Takahiro Kusakabe; Jae Man Lee

doi:10.1016/j.aspen.2019.12.014

Optimal silkworm larva host for high-level production of Mus musculus IL-4 using a baculovirus expression vector system

Masahiko Kobayashi, Jian Xu, Kohei Kakino, Akitsu Masuda, Masato Hino, Naoki Fujimoto, Kosuke Minamihata, Noriho Kamiya, Hiroaki Mon, Hiroshi Iida, Masateru Takahashi, Takahiro Kusakabe, Jae Man Lee

Biological, Environmental Sciences and Engineering

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Interleukine-4 (IL-4) is a cytokine that plays an important role in the immune system and recognized as a biological medicine. Therefore, there is a demand for the production of IL-4 with high performance. The expression of a recombinant IL-4 protein in the prokaryotic system usually results in the formation of an inclusion body. To date, the solution to obtain those active products without the refolding process remains to be established. In this study, we tried to acquire a biologically active recombinant Mus musculus IL-4 (rMmIL-4) using a silkworm-baculovirus expression vector system (silkworm-BEVS). We constructed two recombinant baculoviruses coding rMmIL-4 with the distinct location of affinity purification tags and succeeded in the expression and purification of rMmIL-4 proteins directly without the refolding process. Both purified proteins displayed comparable biological activity to the commercial proteins produced by the E. coli expression system. Besides, we performed screening of silkworm strains to seek optimal hosts for the mass-production of rMmIL-4. Intriguingly, we found that some silkworm strains showed significantly higher secretion levels of rMmIL-4 in silkworm sera. Our study provides meaningful insights into the industrial-scale production of rMmIL-4 with high productivity for pharmaceutical applications in the future.

Original language	English (US)
Pages (from-to)	268-273
Number of pages	6
Journal	Journal of Asia-Pacific Entomology
Volume	23
Issue number	1
DOIs	https://doi.org/10.1016/j.aspen.2019.12.014
State	Published - Dec 31 2019

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: We thank Dr. Imanishi (National Institute of Agrobiological Sciences, Japan) for kindly providing the NIAS-Bm-oyanagi2 (BmO2) cell line. This work was supported by the Japan Science and Technology Agency (JST) for the Program for Creating Start-ups from Advanced Research and Technology (START Program). The authors declare no conflict of interest.

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10.1016/j.aspen.2019.12.014

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Kobayashi, M., Xu, J., Kakino, K., Masuda, A., Hino, M., Fujimoto, N., Minamihata, K., Kamiya, N., Mon, H., Iida, H., Takahashi, M., Kusakabe, T., & Man Lee, J. (2019). Optimal silkworm larva host for high-level production of Mus musculus IL-4 using a baculovirus expression vector system. Journal of Asia-Pacific Entomology, 23(1), 268-273. https://doi.org/10.1016/j.aspen.2019.12.014

@article{bdd67dec4926492a89a34cffbcd6b300,

title = "Optimal silkworm larva host for high-level production of Mus musculus IL-4 using a baculovirus expression vector system",

abstract = "Interleukine-4 (IL-4) is a cytokine that plays an important role in the immune system and recognized as a biological medicine. Therefore, there is a demand for the production of IL-4 with high performance. The expression of a recombinant IL-4 protein in the prokaryotic system usually results in the formation of an inclusion body. To date, the solution to obtain those active products without the refolding process remains to be established. In this study, we tried to acquire a biologically active recombinant Mus musculus IL-4 (rMmIL-4) using a silkworm-baculovirus expression vector system (silkworm-BEVS). We constructed two recombinant baculoviruses coding rMmIL-4 with the distinct location of affinity purification tags and succeeded in the expression and purification of rMmIL-4 proteins directly without the refolding process. Both purified proteins displayed comparable biological activity to the commercial proteins produced by the E. coli expression system. Besides, we performed screening of silkworm strains to seek optimal hosts for the mass-production of rMmIL-4. Intriguingly, we found that some silkworm strains showed significantly higher secretion levels of rMmIL-4 in silkworm sera. Our study provides meaningful insights into the industrial-scale production of rMmIL-4 with high productivity for pharmaceutical applications in the future.",

author = "Masahiko Kobayashi and Jian Xu and Kohei Kakino and Akitsu Masuda and Masato Hino and Naoki Fujimoto and Kosuke Minamihata and Noriho Kamiya and Hiroaki Mon and Hiroshi Iida and Masateru Takahashi and Takahiro Kusakabe and {Man Lee}, Jae",

note = "KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: We thank Dr. Imanishi (National Institute of Agrobiological Sciences, Japan) for kindly providing the NIAS-Bm-oyanagi2 (BmO2) cell line. This work was supported by the Japan Science and Technology Agency (JST) for the Program for Creating Start-ups from Advanced Research and Technology (START Program). The authors declare no conflict of interest.",

year = "2019",

month = dec,

day = "31",

doi = "10.1016/j.aspen.2019.12.014",

language = "English (US)",

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journal = "Journal of Asia-Pacific Entomology",

issn = "1226-8615",

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Kobayashi, M, Xu, J, Kakino, K, Masuda, A, Hino, M, Fujimoto, N, Minamihata, K, Kamiya, N, Mon, H, Iida, H, Takahashi, M, Kusakabe, T & Man Lee, J 2019, 'Optimal silkworm larva host for high-level production of Mus musculus IL-4 using a baculovirus expression vector system', Journal of Asia-Pacific Entomology, vol. 23, no. 1, pp. 268-273. https://doi.org/10.1016/j.aspen.2019.12.014

TY - JOUR

T1 - Optimal silkworm larva host for high-level production of Mus musculus IL-4 using a baculovirus expression vector system

AU - Kobayashi, Masahiko

AU - Xu, Jian

AU - Kakino, Kohei

AU - Masuda, Akitsu

AU - Hino, Masato

AU - Fujimoto, Naoki

AU - Minamihata, Kosuke

AU - Kamiya, Noriho

AU - Mon, Hiroaki

AU - Iida, Hiroshi

AU - Takahashi, Masateru

AU - Kusakabe, Takahiro

AU - Man Lee, Jae

N1 - KAUST Repository Item: Exported on 2020-10-01 Acknowledgements: We thank Dr. Imanishi (National Institute of Agrobiological Sciences, Japan) for kindly providing the NIAS-Bm-oyanagi2 (BmO2) cell line. This work was supported by the Japan Science and Technology Agency (JST) for the Program for Creating Start-ups from Advanced Research and Technology (START Program). The authors declare no conflict of interest.

PY - 2019/12/31

Y1 - 2019/12/31

N2 - Interleukine-4 (IL-4) is a cytokine that plays an important role in the immune system and recognized as a biological medicine. Therefore, there is a demand for the production of IL-4 with high performance. The expression of a recombinant IL-4 protein in the prokaryotic system usually results in the formation of an inclusion body. To date, the solution to obtain those active products without the refolding process remains to be established. In this study, we tried to acquire a biologically active recombinant Mus musculus IL-4 (rMmIL-4) using a silkworm-baculovirus expression vector system (silkworm-BEVS). We constructed two recombinant baculoviruses coding rMmIL-4 with the distinct location of affinity purification tags and succeeded in the expression and purification of rMmIL-4 proteins directly without the refolding process. Both purified proteins displayed comparable biological activity to the commercial proteins produced by the E. coli expression system. Besides, we performed screening of silkworm strains to seek optimal hosts for the mass-production of rMmIL-4. Intriguingly, we found that some silkworm strains showed significantly higher secretion levels of rMmIL-4 in silkworm sera. Our study provides meaningful insights into the industrial-scale production of rMmIL-4 with high productivity for pharmaceutical applications in the future.

AB - Interleukine-4 (IL-4) is a cytokine that plays an important role in the immune system and recognized as a biological medicine. Therefore, there is a demand for the production of IL-4 with high performance. The expression of a recombinant IL-4 protein in the prokaryotic system usually results in the formation of an inclusion body. To date, the solution to obtain those active products without the refolding process remains to be established. In this study, we tried to acquire a biologically active recombinant Mus musculus IL-4 (rMmIL-4) using a silkworm-baculovirus expression vector system (silkworm-BEVS). We constructed two recombinant baculoviruses coding rMmIL-4 with the distinct location of affinity purification tags and succeeded in the expression and purification of rMmIL-4 proteins directly without the refolding process. Both purified proteins displayed comparable biological activity to the commercial proteins produced by the E. coli expression system. Besides, we performed screening of silkworm strains to seek optimal hosts for the mass-production of rMmIL-4. Intriguingly, we found that some silkworm strains showed significantly higher secretion levels of rMmIL-4 in silkworm sera. Our study provides meaningful insights into the industrial-scale production of rMmIL-4 with high productivity for pharmaceutical applications in the future.

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U2 - 10.1016/j.aspen.2019.12.014

DO - 10.1016/j.aspen.2019.12.014

M3 - Article

SN - 1226-8615

VL - 23

SP - 268

EP - 273

JO - Journal of Asia-Pacific Entomology

JF - Journal of Asia-Pacific Entomology

IS - 1

ER -

Optimal silkworm larva host for high-level production of Mus musculus IL-4 using a baculovirus expression vector system

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