TY - JOUR
T1 - Production of an active Mus musculus IL-3 using updated silkworm-based baculovirus expression vector system
AU - Nagai, Ryo
AU - Ebihara, Takeru
AU - Kakino, Kohei
AU - Masuda, Akitsu
AU - Xu, Jian
AU - Minamihata, Kosuke
AU - Kamiya, Noriho
AU - Kongkrongtong, Tatphon
AU - Kawahara, Masahiro
AU - Mon, Hiroaki
AU - Fujii, Tsuguru
AU - Kusakabe, Takahiro
AU - Lee, Jae Man
N1 - Funding Information:
The cultured silkworm BmO2 (NIAS-Bm-oyanagi2) cells were kindly gifted by Dr. Imanishi and stably cultured in IPL-41 medium (Sigma, St. Louis, MO) with 10% fetal bovine serum (Gibco, Grand Island, NY) at 27 °C. The silkworm d17 silkworm larvae ( Kawakami et al., 2008 ) were provided by the Institute of Genetic Resources, Kyushu University Graduate School, supported by the Japan National BioResource Project. The larvae were routinely reared on fresh mulberry leaves under well-controlled environmental conditions at 25–27 °C.
Funding Information:
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).
Publisher Copyright:
© 2021 Korean Society of Applied Entomology
PY - 2021
Y1 - 2021
N2 - Due to the biological significance and therapeutic potential of Interleukin-3 (IL-3) secreted mainly by activated T cells, various protein expression systems have been challenged to produce recombinant IL3 to meet the increasing demands worldwide. Recently, we established an updated silkworm-based baculovirus expression vector system (silkworm-BEVS), which in most cases, produces eukaryotic proteins in biological or enzymatical active forms with considerable amounts. We attempted to reconstruct and express a recombinant mouse IL-3 (rMmIL-3) with C-terminal His8-Strep tags in silkworm-BEVS in the current study. From our results, we gained an active glycosylated rMmIL-3 protein in a substantial amount and quality. As compared with the E. coli expression system, silkworm-BEVS is a better choice regarding the glycosylations attached in rMmIL-3 and up-scalable system in case that a commercial amount is required in the future. Collectively, our method shares an excellent model to produce interleukin molecular for approaching pharmaceutical applications.
AB - Due to the biological significance and therapeutic potential of Interleukin-3 (IL-3) secreted mainly by activated T cells, various protein expression systems have been challenged to produce recombinant IL3 to meet the increasing demands worldwide. Recently, we established an updated silkworm-based baculovirus expression vector system (silkworm-BEVS), which in most cases, produces eukaryotic proteins in biological or enzymatical active forms with considerable amounts. We attempted to reconstruct and express a recombinant mouse IL-3 (rMmIL-3) with C-terminal His8-Strep tags in silkworm-BEVS in the current study. From our results, we gained an active glycosylated rMmIL-3 protein in a substantial amount and quality. As compared with the E. coli expression system, silkworm-BEVS is a better choice regarding the glycosylations attached in rMmIL-3 and up-scalable system in case that a commercial amount is required in the future. Collectively, our method shares an excellent model to produce interleukin molecular for approaching pharmaceutical applications.
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U2 - 10.1016/j.aspen.2021.04.011
DO - 10.1016/j.aspen.2021.04.011
M3 - Article
AN - SCOPUS:85106296770
SN - 1226-8615
VL - 24
SP - 544
EP - 549
JO - Journal of Asia-Pacific Entomology
JF - Journal of Asia-Pacific Entomology
IS - 3
ER -