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.
UR - http://hdl.handle.net/10754/661564
UR - https://linkinghub.elsevier.com/retrieve/pii/S1226861519307277
UR - http://www.scopus.com/inward/record.url?scp=85079222734&partnerID=8YFLogxK
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 -