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Mamestra brassicae NIAS-Mb-32 cell strain 2g2 enables high-yield recombinant protein production in baculovirus-free and baculovirus-based insect cell expression.
Yamamoto, Yotaro; Kajiura, Hiroyuki; Nishibu, Takahiro; Fujiyama, Kazuhito.
  • Yamamoto Y; Production Process Development Department, FUJIFILM Wako Pure Chemical Corporation, 6-1 Takata-cho, Amagasaki-shi, Hyogo 661-0963, Japan; International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan.
  • Kajiura H; International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan.
  • Nishibu T; Production Process Development Department, FUJIFILM Wako Pure Chemical Corporation, 6-1 Takata-cho, Amagasaki-shi, Hyogo 661-0963, Japan.
  • Fujiyama K; International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan. Electronic address: fujiyama@icb.osaka-u.ac.jp.
J Biosci Bioeng ; 134(5): 432-440, 2022 Nov.
Article in English | MEDLINE | ID: covidwho-2041897
ABSTRACT
The production of recombinant proteins using insect cells has been widely used for over 30 years, which contributing to life science research and biotechnology. Insect cells exhibiting enhanced N-glycosylation and recombinant protein productivity enhance the productivity of the baculovirus-insect cell system (BICS). A new highly proliferative insect cell strain, 2g2, was established from the Mamestra brassicae pupa ovary cell strain NIAS-MB-32 (RCB0413) to address the problem of Sf-rhabdovirus and to explore the newly available possibilities in BICS as well as Sf9, such as increased protein production and recombinant baculovirus amplification. The high-growth cell strain 2g2 was examined for its recombinant protein production ability and baculovirus productivity; moreover, the activity of the produced recombinant proteins was examined using Sf9 as a benchmark. Recombinant protein productivity and virus production by BICS in 2g2 was confirmed as equivalent to that of Sf9. Furthermore, we produced the severe acute respiratory syndrome coronavirus 2 spike protein in a baculovirus-free system and compared its productivity, binding activity with human angiotensin-converting enzyme 2, and N-glycosylation. The productivity and bioactivity were found to be equal to or better than that of Sf9. Moreover, N-glycosylation analysis revealed that the glycans derived from the 2g2-produced glycoproteins were mostly of the high mannose type as Sf9. Therefore, 2g2 may have the same N-glycosylation ability as Sf9. Finally, the Sf-rhabdovirus was confirmed to be negative in 2g2. Our results demonstrated that the novel insect cell strain 2g2 can serve as a protein production tool in scientific research and industrial biotechnology.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Baculoviridae / COVID-19 Limits: Animals / Humans Language: English Journal: J Biosci Bioeng Journal subject: Biomedical Engineering / Microbiology Year: 2022 Document Type: Article Affiliation country: J.jbiosc.2022.07.010

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Baculoviridae / COVID-19 Limits: Animals / Humans Language: English Journal: J Biosci Bioeng Journal subject: Biomedical Engineering / Microbiology Year: 2022 Document Type: Article Affiliation country: J.jbiosc.2022.07.010