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Novel assay for determination of baculovirus titer using host cell binding of enzyme-displayed baculovirus

Research Project

Project/Area Number 21560805
Research Category

Grant-in-Aid for Scientific Research (C)

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

Principal Investigator

KIKIUCHI Ken-ichi  秋田大学, 大学院・工学資源学研究科, 教授 (80108919)

Co-Investigator(Kenkyū-buntansha) GOTOH Takeshi  秋田大学, 大学院・工学資源学研究科, 教授 (10215494)
Project Period (FY) 2009 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2010: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2009: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Keywords生物機能工学 / 生体認識 / ウイルス / バイオテクノロジー / 昆虫細胞 / バキュロウイルス / 酵素反応 / リアルタイムPCR / ウイルス検出 / ウイルス力価 / 酵素展示
Research Abstract

The present study aimed at developing a novel assay method using a baculovirus (BV)-host cell binding for the determination of BV titer. First, we constructed recombinant BV displaying either GST or EGFP on GP64 envelope protein in order to distinguish between infected and uninfected cells. However, the protein probes displayed on BVs were not sensitive enough to use for this purpose. In addition, it was so difficult to produce the recombinant BVs because of their high susceptibility to endogenous proteases. Second, we examined the BV-host cell binding conditions to separate active and inactive BVs. As a result, it was suggested that the BV-host cell binding was applicable to the BV titer determination by combining with real-time PCR.

Report

(4 results)
  • 2011 Annual Research Report   Final Research Report ( PDF )
  • 2010 Annual Research Report
  • 2009 Annual Research Report
  • Research Products

    (8 results)

All 2012 2010 2009

All Journal Article (4 results) (of which Peer Reviewed: 2 results) Presentation (4 results)

  • [Journal Article] Purification and characterization of aspartic protease derived from Sf9 insect cells.2010

    • Author(s)
      Gotoh, T., Ono, H., Kikuchi, K.-I., Nirasawa, S., Takahashi, S.
    • Journal Title

      Biosci Biotechnol Biochem.

      Volume: 74 Pages: 2154-2157

    • NAID

      10027561012

    • Related Report
      2011 Final Research Report
  • [Journal Article] Prorenin processing enzyme (PPE) produced by Baculovirus-infected Sf-9 insect cells: PPE is the cysteine protease encoded in the AcMNPV gene2010

    • Author(s)
      Gotoh, T., Awa, H., Kikuchi, K.-I., Nirasawa, S., Takahashi, S.
    • Journal Title

      Biosci Biotechnol Biochem.

      Volume: 74 Pages: 70-74

    • NAID

      10027551660

    • Related Report
      2011 Final Research Report
  • [Journal Article] Prorenin processing enzyme (PPE) produced by baculovirus-infected Sf-9insect cells : PPE is the cysteine protease encoded in the AcMNPV gene2010

    • Author(s)
      Takeshi Gotoh, 他4名
    • Journal Title

      Biosci.Biotechnol.Biochem.

      Volume: 74 Pages: 370-374

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Purification and characterization of aspartic protease derived from Sf9 insect cells2010

    • Author(s)
      Takeshi Gotoh, 他4名
    • Journal Title

      Biosci.Biotechnol.Biochem.

      Volume: 74 Pages: 2154-2157

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] Real-time PCRを利用したバキュロウイルスタイター測定のための基礎検討2012

    • Author(s)
      佐藤 真一, 佐藤 愛香, 菊地 賢一, 後藤 猛
    • Organizer
      日本化学工学会第77年会
    • Place of Presentation
      工学院大学
    • Year and Date
      2012-03-15
    • Related Report
      2011 Annual Research Report 2011 Final Research Report
  • [Presentation] GP64-蛍光プローブ融合タンパク質の構築とSf-9昆虫細胞への結合2009

    • Author(s)
      村上 亮, 後藤 猛, 菊地 賢一
    • Organizer
      第61回日本生物工学会大会
    • Place of Presentation
      名古屋大学
    • Year and Date
      2009-09-24
    • Related Report
      2011 Final Research Report 2009 Annual Research Report
  • [Presentation] 蛍光タンパク質融合GP64の構築と昆虫細胞への結合性2009

    • Author(s)
      村上亮, 後藤猛, 菊地賢一
    • Organizer
      第9回食品酵素科学研究会・第15回秋田応用生命科学研究会学術講演会
    • Place of Presentation
      秋田県総合食品研究所
    • Year and Date
      2009-09-04
    • Related Report
      2009 Annual Research Report
  • [Presentation] 蛍光タンパク質融合GP64 の構築と昆虫細胞への結合性2009

    • Author(s)
      村上 亮, 後藤 猛, 菊地 賢一
    • Organizer
      第9回食品酵素化学研究会・第15回秋田応用生命科学研究会学術講演会
    • Place of Presentation
      秋田県総合食品研究センター
    • Related Report
      2011 Final Research Report

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Published: 2009-04-01   Modified: 2016-04-21  

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