Continuous production of virus-like particles by recombinant insect cells

• Project/Area Number: 19560777
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Kobe University
• Principal Investigator: YAMAJI Hideki Kobe University, 大学院工学研究科, 准教授 (40283874)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2008: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2007: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: 昆虫細胞 / 組換えタンパク質生産 / ウイルスタンパク質 / ワクチン / ウイルス様粒子 / 日本脳炎ウイルス / 遺伝子組換え

Research Abstract

ウイルスの表面タンパク質の遺伝子を発現する組換え昆虫細胞を作製し, 安全かつ有効なワクチンとして利用可能なウイルス様粒子の生産について検討した. カイコガ由来のアクチンプロモーターの上流にバキュロウイルス由来のトランス作用因子とエンハンサーを配し, 選択マーカーを有する発現ベクターに, 日本脳炎ウイルスの表面タンパク質prMおよびEの遺伝子を挿入した. このプラスミドを培養昆虫細胞に導入して薬剤存在下で培養することにより, 哺乳動物細胞と比べて100倍以上のEタンパク質を分泌生産する細胞を得ることに成功した.

Research Products

• [Journal Article] A high-level expression vector containing selectable marker for continuous production of recombinant protein in insect cells (2009)
  • Author(s): Takuya Shishido, Naoya Kurata, Myung Eui Yoon, Tsutomu Tanaka, Hideki Yamaji, Hideki Fukuda, Akihiko Kondo
  • Journal Title: Biotechnol. Lett. 31 Pages: 623-627
  • Peer Reviewed
• [Journal Article] Production of functional antibody Fab fragment by recombinant insect cells (2008)
  • Author(s): Hideki Yamaji, Toshitaka Manabe, Keizo Watakabe, Masaru Muraoka, Ikuo Fujii, Hideki Fukuda
  • Journal Title: Biochem. Eng. J. 41 Pages: 203-209
  • NAID: 120001478904
  • Peer Reviewed
• [Journal Article] Production of bionanocapsules in immobilized insect cell culture using porous biomass support particles (2007)
  • Author(s): Takuya Shishido, Masaru Muraoka, Hideki Yamaji, Akihiko Kondo, Hideki Fukuda
  • Journal Title: J. Biosci. Bioeng. 103 Pages: 572-574
  • NAID: 110006385794
  • Peer Reviewed
• [Presentation] 昆虫細胞を用いた日本脳炎ウイルスタンパク質の生産 (2009)
  • Author(s): 高橋裕輔, 中村匡崇, 勝田知尚, 山地秀樹
  • Organizer: 化学工学会第74年会
  • Place of Presentation: 横浜
  • Year and Date: 2009-03-19
• [Presentation] 昆虫細胞を用いた日木脳炎ウイルスタンパク質の生産 (2009)
  • Author(s): 高橋 裕輔, 他
  • Organizer: 化学工学会第74年会
  • Place of Presentation: 横浜国立大学
  • Year and Date: 2009-03-19
• [Presentation] Low multiplicity infection of 293 cells for adenovirus vector production (2008)
  • Author(s): Naoya Morishita, Kentaro Yamada, Shuji Kubo, Akinobu Gotoh, Tomohisa Katsuda, Hideki Fukuda, Hideki Yamaji
  • Organizer: JAACT 2008 Fukuoka (The 21th Annual and International Meeting of the Japanese Association for Animal Cell Technology), "POWER OF CELLS, Animal Cell Technology in Bioscience, Bioengineering and Bioindustries"
  • Place of Presentation: Fukuoka
  • Year and Date: 2008-11-26
• [Presentation] Production of adenovirus vector by 293 cells immobilized within porous biomass support particles (2008)
  • Author(s): Naoya Morishita, Shuji Kubo, Akinobu Gotoh, Tomohisa Katsuda, Hideki Fukuda, Hideki Yamaji
  • Organizer: JAACT 2008 Fukuoka (The 21th Annual and International Meeting of the Japanese Association for Animal Cell Technology), "POWER OF CELLS Animal, Cell Technology in Bioscience, Bioengineering and Bioindustries"
  • Place of Presentation: Fukuoka
  • Year and Date: 2008-11-26
• [Presentation] Secretory production of Japanese encephalitis virus E protein by recombinant insect cells (2008)
  • Author(s): Yusuke Takahashi, Masataka Nakamura, Takanori Furuta, Tomohisa Katsuda, Hideki Yamaji
  • Organizer: "Bioseparation for Biorecognition and Bionanotechnology" Conference
  • Place of Presentation: Hanyang University, Korea
  • Year and Date: 2008-10-27
• [Presentation] Secretory produclion of Japanese cnccphalilis virus E protcin by recombinant insect cells (2008)
  • Author(s): Y. Takahashi, et al.
  • Organizer: “Bioseparation for Biorccognition and Bionanotech-nology" Conference
  • Place of Presentation: Hanyang University, Ansan, Korea
  • Year and Date: 2008-10-27
• [Presentation] 293細胞の固定化培養によるアデノウイルスベクターの生産 (2008)
  • Author(s): 森下直矢, 山田健太郎, 山地秀樹, 福田秀樹
  • Organizer: 化学工学会第73年会
  • Place of Presentation: 浜松
  • Year and Date: 2008-03-18
• [Presentation] Display of functional antibody Fab fragment on the baculovirus surface (2007)
  • Author(s): Hideki Yamaji, Hideki Fukuda
  • Organizer: 13th European Congress on Biotechnology
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2007-09-18
• [Presentation] 昆虫細胞を用いる機能性抗体の生産システム (2007)
  • Author(s): 古田貴憲, 山地秀樹, 福田秀樹
  • Organizer: 化学工学会第39回秋季大会
  • Place of Presentation: 札幌
  • Year and Date: 2007-09-14