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Production of influenza virus-like particles in recombinant insect cells

Research Project

Project/Area Number 15H04195
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biofunction/Bioprocess
Research InstitutionKobe University

Principal Investigator

Yamaji Hideki  神戸大学, 工学研究科, 教授 (40283874)

Research Collaborator KATSUDA Tomohisa  
Project Period (FY) 2015-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000)
Fiscal Year 2018: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
Fiscal Year 2017: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
Fiscal Year 2016: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2015: ¥6,110,000 (Direct Cost: ¥4,700,000、Indirect Cost: ¥1,410,000)
Keywords昆虫細胞 / 組換えタンパク質生産 / ウイルス様粒子 / インフルエンザウイルス / ワクチン / 生物化学工学
Outline of Final Research Achievements

Production of influenza virus-like particles in recombinant insect cells was investigated. The cDNA fragments encoding hemagglutinin (HA) and matrix protein 1 (M1) of an influenza virus A were separately cloned into the plasmid vector pIHAbla and pIHAneo. The pIHAbla and pIHAneo contained the Bombyx mori actin promoter downstream of the B. mori nucleopolyhedrovirus (BmNPV) IE-1 transactivator and the BmNPV HR3 enhancer for high-level expression, together with either a blasticidin or neomycin resistance gene for use as a selectable marker, respectively. After cotransfection with the prepared plasmids, High Five cells were incubated with blasticidin and G418, and cells resistant to the antibiotics were effectively obtained. Sucrose density-gradient sedimentation analysis of the culture supernatant showed that secreted HA and M1 molecules were produced in a particulate form.

Academic Significance and Societal Importance of the Research Achievements

複数の構造タンパク質とエンベロープから構成されるインフルエンザウイルス様粒子の組換え昆虫細胞による生産については,これまで報告されていない.本研究で構築した基盤技術は,亜型の異なるインフルエンザウイルスのウイルス様粒子ワクチンを開発・生産する際にも利用することができる.気候変動,グローバル化,高齢化などにより感染症の深刻化が懸念される状況のもと,本研究で得られた成果はインフルエンザなどのウイルス感染症から人類を守る一助になると期待される.

Report

(5 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Annual Research Report
  • 2016 Annual Research Report
  • 2015 Annual Research Report
  • Research Products

    (25 results)

All 2019 2018 2017 2016 Other

All Int'l Joint Research (1 results) Journal Article (7 results) (of which Peer Reviewed: 4 results,  Open Access: 2 results) Presentation (15 results) (of which Int'l Joint Research: 10 results) Book (2 results)

  • [Int'l Joint Research] University of Malaya(Malaysia)

    • Related Report
      2015 Annual Research Report
  • [Journal Article] Effects of lithium on the secretory production of recombinant antibody from insect cells2019

    • Author(s)
      Yuki Ohmuro-Matsuyama, Tomohisa Katsuda, Hideki Yamaji
    • Journal Title

      In Vitro Cellular & Developmental Biology - Animal

      Volume: 55 Issue: 1 Pages: 1-6

    • DOI

      10.1007/s11626-018-0303-1

    • NAID

      120006601532

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Modifications of a signal sequence for antibody secretion from insect cells2018

    • Author(s)
      Yuki Ohmuro-Matsuyama, Hideki Yamaji
    • Journal Title

      Cytotechnology

      Volume: 70 Issue: 3 Pages: 891-898

    • DOI

      10.1007/s10616-017-0109-0

    • NAID

      120006539286

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Production of influenza virus proteins in stably transformed insect cells2018

    • Author(s)
      Hideki Yamaji, Toshikazu Tanijima, Takuya Matsuda, Kyoko Masumi-Koizumi, Tomohisa Katsuda
    • Journal Title

      New Biotechnology

      Volume: 44 Pages: S159-S159

    • DOI

      10.1016/j.nbt.2018.05.1167

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Efficient protein production by transient gene expression using insect cells2018

    • Author(s)
      Hideki Yamaji, Keita Mori, Hirotsugu Hamada, Yuki Ohmuro-Matsuyama, Tomohisa Katsuda
    • Journal Title

      BMC Proceedings

      Volume: 12 Issue: S1 Pages: 19-19

    • DOI

      10.1186/s12919-018-0097-x

    • Related Report
      2017 Annual Research Report
    • Open Access
  • [Journal Article] Efficient production of antibody Fab fragment by transient gene expression in insect cells2017

    • Author(s)
      Keita Mori, Hirotsugu Hamada, Takafumi Ogawa, Yuki Ohmuro-Matsuyama, Tomohisa Katsuda, and Hideki Yamaji
    • Journal Title

      Journal of Bioscience and Bioengineering

      Volume: - Issue: 2 Pages: 221-226

    • DOI

      10.1016/j.jbiosc.2017.03.007

    • NAID

      120006425709

    • Related Report
      2017 Annual Research Report 2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Electrostatic engineering of the interface between heavy and light chains promotes antibody Fab fragment production2016

    • Author(s)
      Ohmuro-Matsuyama Yuki、Mori Keita、Hamada Hirotsugu、Ueda Hiroshi、Yamaji Hideki
    • Journal Title

      Cytotechnology

      Volume: 69 Issue: 3 Pages: 469-475

    • DOI

      10.1007/s10616-016-9955-4

    • NAID

      120006323719

    • Related Report
      2017 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 昆虫細胞を用いた有用物質生産2016

    • Author(s)
      山地 秀樹
    • Journal Title

      生物工学会誌

      Volume: 94 Pages: 76-80

    • Related Report
      2015 Annual Research Report
    • Open Access
  • [Presentation] Production of influenza virus-like particles in insect cells2019

    • Author(s)
      Hideki Yamaji, Takuya Matsuda, Toshikazu Tanijima, Kyoko Masumi-Koizumi, Tomohisa Katsuda
    • Organizer
      26th ESACT Meeting (ESACT 2019)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Production of influenza virus proteins in recombinant insect cells2019

    • Author(s)
      Takuya Matsuda, Toshikazu Tanijima, Kyoko Masumi-Koizumi, Tomohisa Katsuda, Hideki Yamaji
    • Organizer
      18th Asian Pacific Confedration of Chemical Engineering Congress (APCChE 2019)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Production of influenza virus proteins in stably transformed insect cells2018

    • Author(s)
      Hideki Yamaji, Toshikazu Tanijima, Takuya Matsuda, Kyoko Masumi-Koizumi, Tomohisa Katsuda
    • Organizer
      18th European Congress on Biotechnology (ECB2018)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 昆虫細胞における2Aペプチドを用いた抗体生産2018

    • Author(s)
      溝手結,増見恭子,勝田知尚,山地秀樹
    • Organizer
      第70回日本生物工学会大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] ゲノム編集技術を用いた組換え昆虫細胞の作製2018

    • Author(s)
      戸上真也,増見恭子,勝田知尚,山地秀樹
    • Organizer
      化学工学会第50回秋季大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Production of influenza virus-like particles in stably transformed insect cells2018

    • Author(s)
      Takuya Matsuda, Toshikazu Tanijima, Kyoko Masumi-Koizumi, Tomohisa Katsuda, Hideki Yamaji
    • Organizer
      31st Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2018 Tsukuba)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Production of antibody Fab fragment using 2A peptide in insect cells2018

    • Author(s)
      Yu Mizote, Kyoko Masumi-Koizumi, Tomohisa Katsuda, Hideki Yamaji
    • Organizer
      31st Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2018 Tsukuba)
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 昆虫細胞によるインフルエンザウイルス抗原タンパク質の生産2018

    • Author(s)
      松田拓也,谷島寿和,増見恭子,勝田知尚,山地秀樹
    • Organizer
      化学工学会第83年会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Efficient protein production by transient gene expression using insect cells2017

    • Author(s)
      Hideki Yamaji, Keita Mori, Hirotsugu Hamada, Yuki Ohmuro-Matsuyama, Tomohisa Katsuda
    • Organizer
      The 25th ESACT Meeting (ESACT 2017)
    • Place of Presentation
      Lausanne, Switherland
    • Year and Date
      2017-05-14
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Efficient protein production by transient gene expression using insect cells2017

    • Author(s)
      Hideki Yamaji, Keita Mori, Hirotsugu Hamada, Yuki Ohmuro-Matsuyama, Tomohisa Katsuda
    • Organizer
      25th ESACT Meeting (ESACT 2017)
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 昆虫細胞を用いたインフルエンザウイルスタンパク質の生産2017

    • Author(s)
      山地秀樹,谷島寿和,増見恭子,勝田知尚
    • Organizer
      第30回日本動物細胞工学会2017年度大会 (JAACT2017)
    • Related Report
      2017 Annual Research Report
  • [Presentation] Production of influenza virus proteins by recombinant insect cells2017

    • Author(s)
      Hideki Yamaji, Toshikazu Tanishima, Kyoko Masumi-Koizumi, Tomohisa Katsuda
    • Organizer
      International Meeting & 42nd Annual Conference of the Malaysian Society for Biochemistry & Molecular Biology (MSBMB)
    • Related Report
      2017 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 昆虫細胞による2Aペプチドを用いた抗体タンパク質の生産2017

    • Author(s)
      溝手結,増見恭子,勝田知尚,山地秀樹
    • Organizer
      化学工学会第49回秋季大会
    • Related Report
      2017 Annual Research Report
  • [Presentation] Production of virus-like particles using recombinant insect cells2016

    • Author(s)
      Toshikazu Tanishima, Tomohisa Katsuda, Hideki Yamaji
    • Organizer
      The 29th Annual and International Meeting of the Japanese Association for Animal Cell Technology (JAACT2016 Kobe)
    • Place of Presentation
      神戸
    • Year and Date
      2016-11-09
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Production of virus-like particles using insect cells2016

    • Author(s)
      Hideki Yamaji
    • Organizer
      The 2016 World Congress on In Vitro Biology
    • Place of Presentation
      San Diego, USA
    • Year and Date
      2016-06-11
    • Related Report
      2016 Annual Research Report 2015 Annual Research Report
    • Int'l Joint Research
  • [Book] バイオロジクスの開発と品質・安全性確保 上巻2018

    • Author(s)
      山地 秀樹
    • Publisher
      (株) エル・アイ・シー
    • ISBN
      9784900487567
    • Related Report
      2018 Annual Research Report
  • [Book] Sunil Thomas (ed.), Vaccine Design: Methods and Protocols, Volume 2: Vaccines for Veterinary Diseases, Methods in Molecular Biology, vol. 14042016

    • Author(s)
      Hideki Yamaji, Eiji Konishi
    • Publisher
      Springer Science+Business Media
    • Related Report
      2015 Annual Research Report

URL: 

Published: 2015-04-16   Modified: 2020-03-30  

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