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PEG化リン脂質ミセルによる糖鎖電気泳動分析の高性能化

Research Project

Project/Area Number 09J00160
Research Category

Grant-in-Aid for JSPS Fellows

Allocation TypeSingle-year Grants
Section国内
Research Field Analytical chemistry
Research InstitutionKyoto University

Principal Investigator

川井 隆之  京都大学, 工学研究科, 特別研究員(DC1)

Project Period (FY) 2009 – 2011
Project Status Completed (Fiscal Year 2011)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2011: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2010: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2009: ¥700,000 (Direct Cost: ¥700,000)
Keywords糖鎖分析 / キャピラリー電気泳動 / マイクロチップ電気泳動 / オンライン試料濃縮 / 光学異性体分析 / 内面修飾法
Research Abstract

large-volume sample stacking with electroosmotic flow pump (LVSEP)法を用いる高感度電気泳動分析法をアニオン性,カチオン性の両試料に適用し,動電クロマトグラフィー(EKC)などの様々な分離モードで分析することに成功し,採用第2年度までに最大で1000倍程度の高感度化に成功した。第3年度では,さらなる高感度化を目標とし,field-amplified sample injection (FASI)法とLVSEP法との結合について検討を行った。
LVSEP-FASI法の濃縮性能を検討するため,モデル試料としてfluoresceinを用いてキャピラリー電気泳動(CE)をベースにLVSEP-FASI分析を行った。バイアルの交換時間および印加圧力の最適化の結果,それぞれ10 min,1.0 psiとした際従来のCE分析と比較して70,000倍の高感度化が達成された。向様にAPTS誘導体化G7のWSEP-FASI-CE分析を行った結果,0.5 psiの圧力印加および15 minにおけるバイアルの交換により,CEと比較して10,000倍の高感度化および未反応のAPTSとG7のベースライン分離が達成された。検出時間,ピーク高さのRSDはそれぞれ2.4%,11%であり,オンライン試料濃縮を長時間行う分析法としては良好な再現性が得られた。また,分離性能の最適化の結果,
LVSEP同様にLVSEP-FASI法を用いてもほとんど分離度が損なわれないことが判明した。
同様にマイクロチップ電気泳動(MCE)においてもLVSEP-FASIの検討を行った。モデル試料としてfluoresceinを用い,試料濃縮時間を240sとしてLVSEP-FASI-MCE分析を行ったところ,わずか10pMの試料の検出に成功した。従来法であるpinched injectionを用いたMCE分析の結果と比較したところ,6万倍の高感度化が達成された。
以上のように,CE,MCEのどちらにおいてもLVSEP-FASI法の高い濃縮性能分離性能が確認された。

Current Status of Research Progress
Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

当初の研究計画とは異なる方向での研究成果ではあるが,糖鎖分析における高感度化,分析操作の簡略化など,高性能化は計画以上に達成されている。また,別途推進している新規親水性ナノ材料を用いた分離性能の向上,開発したLVSEP法と質量分析計(MS)との結合などについても順調に推移しており,現在開発された新規高性能糖鎖分析法の実分析への応用を推進中である。

Strategy for Future Research Activity

開発した高性能糖鎖電気泳動法とMSとを結合し,糖鎖実試料分析へ適用する。既に開発したLVSEP法とMSの結合については検証済みであり,今後この技術を用いて糖タンパク質の解析を実行する予定である。

Report

(3 results)
  • 2011 Annual Research Report
  • 2010 Annual Research Report
  • 2009 Annual Research Report
  • Research Products

    (33 results)

All 2012 2011 2010 2009

All Journal Article (6 results) (of which Peer Reviewed: 6 results) Presentation (27 results)

  • [Journal Article] Highly sensitive oligosaccharide analysis in capillary electrophoresis using large-volume sample stacking with an electroosmotic flow pump2012

    • Author(s)
      Takayuki Kawai, Masato Watanabe, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Journal Title

      Journal of Chromatography A

      Volume: 1,232 Pages: 52-58

    • DOI

      10.1016/j.chroma.2011.09.032

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Recent Progress of On-line Sample Preconcentration Techniques in Microchip Electrophoresis2012

    • Author(s)
      Fumihiko Kitagawa, Takayuki Kawai, Kenji Sueyoshi, Koji Otsuka
    • Journal Title

      Analytical Sciences

      Volume: 28 Issue: 2 Pages: 85

    • DOI

      10.2116/analsci.28.85

    • NAID

      130004827205

    • ISSN
      0910-6340, 1348-2246
    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Highly sensitive chiral analysis in capillary electrophoresis with large-volume sample stacking with an electroosmotic flow pump2012

    • Author(s)
      Takayuki Kawai, Hiroshi Koino, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Journal Title

      Journal of Chromatography A

      Volume: (印刷中) Pages: 28-34

    • DOI

      10.1016/j.chroma.2012.02.001

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Hydrophobic labeling of amino acids : Transient trapping-capillary/microchip electrophoresis2011

    • Author(s)
      Kenji Sueyoshi, Kota Hashiba, Takayuki Kawai, Fumihiko Kitagawa, Koji Otsuka
    • Journal Title

      Electrophoresis

      Volume: 32 Issue: 10 Pages: 1233-1240

    • DOI

      10.1002/elps.201000567

    • Related Report
      2011 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Hydrophobic labeling of amino acids : Transient trapping-capill ary/microchip electrophoresis2011

    • Author(s)
      Kenji Sueyoshi, Kota Hashiba, Takayuki Kawai, Fumihiko Kitagawa, Koji Otsuka
    • Journal Title

      Electrophoresis

      Volume: 32 Pages: 1233-1240

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Microchip electrophoresis of oligosaccharides using large-volume sample stacking with electroosmotic flow pump in single channel2010

    • Author(s)
      Takayuki Kawai, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Journal Title

      Analytical Chemistry

      Volume: 82 Pages: 6504-6511

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Presentation] Toward 10,000-fold sensitivity improvement in capillary electrophoresis of oligosaccharides2011

    • Author(s)
      Takayuki Kawai, Masumi Ueda, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      11th Asia-Pacific International Symposium on Microscale Separation and Analysis
    • Place of Presentation
      Hobart, Australia
    • Year and Date
      2011-11-29
    • Related Report
      2011 Annual Research Report
  • [Presentation] LVSEP-FASI法を用いた糖鎖電気泳動分析の1万倍の高感度化(3)2011

    • Author(s)
      川井隆之, 上田真澄, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      第31回キャピラリー電気泳動シンポジウム
    • Place of Presentation
      鶴岡
    • Year and Date
      2011-11-09
    • Related Report
      2011 Annual Research Report
  • [Presentation] LVSEP-FASI法を用いた糖鎖電気泳動分析の1万倍の高感度化(2)2011

    • Author(s)
      川井隆之, 上田真澄, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      第22回クロマトグラフィー科学会議
    • Place of Presentation
      仙台
    • Year and Date
      2011-10-22
    • Related Report
      2011 Annual Research Report
  • [Presentation] Toward 10,000-fold sample preconcentration efficiency for oligosaccharide analysis in capillary electrophoresis2011

    • Author(s)
      Takayuki Kawai, Masumi Ueda, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      37th International Symposium on High Performance Liquid Phase Separations and Related Techniques
    • Place of Presentation
      Dalian, China
    • Year and Date
      2011-10-10
    • Related Report
      2011 Annual Research Report
  • [Presentation] Over 50,000-fold sample preconcentration efficiency in microchip elctrophoresis using a simple channel2011

    • Author(s)
      Takayuki Kawai, Masumi Ueda, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      15th International Conference on Miniaturized Systems for Chemistry and Life Sciences
    • Place of Presentation
      Seattle, DSA
    • Year and Date
      2011-10-05
    • Related Report
      2011 Annual Research Report
  • [Presentation] LVSEP-FASI法を用いた糖鎖電気泳動分析の1万倍の高感度化2011

    • Author(s)
      川井隆之, 上田真澄, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      日本分析化学会第60年会
    • Place of Presentation
      名古屋
    • Year and Date
      2011-09-14
    • Related Report
      2011 Annual Research Report
  • [Presentation] Toward 10,000-fold Sensitivity Enhancement for Oligosaccharide Analysis in Capillary Electrophoresis. 22011

    • Author(s)
      Takayuki Kawai, Masumi Ueda, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      JAIMA Conference 2011
    • Place of Presentation
      千葉
    • Year and Date
      2011-09-08
    • Related Report
      2011 Annual Research Report
  • [Presentation] Highly sensitive chiral analysis in microscale electrophoresis using large-volume sample stacking with electroosmotic flow pump2011

    • Author(s)
      Takayuki Kawai, Jun Ito, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      36th International Symposium on High Performance Liquid Phase Separations and Related Techniques
    • Place of Presentation
      Budapest, Hungary
    • Year and Date
      2011-06-22
    • Related Report
      2011 Annual Research Report
  • [Presentation] FASI法およびLVSEP法を用いた高感度マイクロチップ電気泳動法の開発2011

    • Author(s)
      川井隆之, 上田真澄, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      第23回化学とマイクロ・ナノシステム研究会
    • Place of Presentation
      千葉
    • Year and Date
      2011-06-11
    • Related Report
      2011 Annual Research Report
  • [Presentation] LVSEP法を用いた光学異性体電気泳動分析の高感度化2011

    • Author(s)
      川井隆之, 伊藤潤, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      第18回クロマトグラフィーシンポジウム
    • Place of Presentation
      福岡
    • Year and Date
      2011-06-03
    • Related Report
      2011 Annual Research Report
  • [Presentation] Highly sensitive chiral analysis by capillary electrophoresis using large-volume sample stacking with electroosmotic flow pump2011

    • Author(s)
      Takayuki Kawai, Jun Ito, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      IUPAC International Congress on Analytical Sciences 2011
    • Place of Presentation
      Kyoto, Japan
    • Year and Date
      2011-05-25
    • Related Report
      2011 Annual Research Report
  • [Presentation] Highly sensitive chiral analysis in cappillary eletropheoresis using large-volume sample stacking with electroosmotic flow pump2011

    • Author(s)
      Takayuki Kawai, Jun Ito, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      26th International Symposium on Microscale Bioseparations
    • Place of Presentation
      San Diego, USA
    • Year and Date
      2011-05-04
    • Related Report
      2011 Annual Research Report
  • [Presentation] LVSEP法によるミクロスケール電気泳動の高感度化(3)2011

    • Author(s)
      川井隆之, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      日本化学会第91春期年会
    • Place of Presentation
      横浜
    • Year and Date
      2011-03-27
    • Related Report
      2010 Annual Research Report
  • [Presentation] Microchip electrophoresis of oligosaccharides with large-volume sample stacking with electroosmotic flow pump in single channel2010

    • Author(s)
      Takayuki Kawai, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      2010 International Chemical Congress of Pacific Basin Societies
    • Place of Presentation
      Honolulu, U.S.
    • Year and Date
      2010-12-16
    • Related Report
      2010 Annual Research Report
  • [Presentation] LVSEP法によるミクロスケール電気泳動の高感度化(2)2010

    • Author(s)
      川井隆之, 木南冴子, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      第30回キャピラリー電気泳動シンポジウム
    • Place of Presentation
      岐阜
    • Year and Date
      2010-11-15
    • Related Report
      2010 Annual Research Report
  • [Presentation] Microchip electrophoresis of oligosaccharides in 'single' straight channel2010

    • Author(s)
      Takayuki Kawai, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      14th International Conference on Miniaturized Systems for Chemistry and Life Sciences
    • Place of Presentation
      Groningen, Netherland
    • Year and Date
      2010-10-06
    • Related Report
      2010 Annual Research Report
  • [Presentation] LVSEP法によるミクロスケール電気泳動の高感度化2010

    • Author(s)
      川井隆之, 木南冴子, 末吉健志, 北川文彦, 大塚浩二
    • Organizer
      日本分析化学会第59年会
    • Place of Presentation
      仙台
    • Year and Date
      2010-09-15
    • Related Report
      2010 Annual Research Report
  • [Presentation] Microchip electrophoresis of sugar chains with large-volume sample stacking in single integrated channel2010

    • Author(s)
      Takayuki Kawai, Kenji Sueyoshi, Fumihiko Kitagawa, Koji Otsuka
    • Organizer
      35th International Symposium on High-Performance Liquid Phase Separation and Related Techniques
    • Place of Presentation
      Boston, U.S.
    • Year and Date
      2010-07-01
    • Related Report
      2010 Annual Research Report
  • [Presentation] Highly Sensitive Electrophoretic Analyses of Sugar Chains on Straight Channel Microchip. 22010

    • Author(s)
      川井隆之
    • Organizer
      25th International Symposium on Microscale Bioseparations
    • Place of Presentation
      Prague, Czech Republic
    • Year and Date
      2010-03-24
    • Related Report
      2009 Annual Research Report
  • [Presentation] キャピラリー/マイクロチップ電気泳動における糖鎖分析の高感度化(2)2009

    • Author(s)
      川井隆之
    • Organizer
      第29回キャピラリー電気泳動シンポジウム
    • Place of Presentation
      東大阪
    • Year and Date
      2009-11-17
    • Related Report
      2009 Annual Research Report
  • [Presentation] キャピラリー/マイクロチップ電気泳動における糖鎖分析の高感度化2009

    • Author(s)
      川井隆之
    • Organizer
      クロマトグラフィー科学会議
    • Place of Presentation
      東京
    • Year and Date
      2009-11-11
    • Related Report
      2009 Annual Research Report
  • [Presentation] Highly Sensitive Electrophoretic Analyses of Sugar Chains on Straight Channel Microchip2009

    • Author(s)
      川井隆之
    • Organizer
      20^<th> CHEMINAS & 10^<th> ISMM
    • Place of Presentation
      金沢
    • Year and Date
      2009-11-07
    • Related Report
      2009 Annual Research Report
  • [Presentation] キャピラリー電気泳動における糖鎖分析の高感度化(4)2009

    • Author(s)
      川井隆之
    • Organizer
      分析化学会第58年会
    • Place of Presentation
      札幌
    • Year and Date
      2009-09-24
    • Related Report
      2009 Annual Research Report
  • [Presentation] キャピラリー電気泳動における糖鎖分析の高感度化(5)2009

    • Author(s)
      川井隆之
    • Organizer
      東京コンファレンス
    • Place of Presentation
      東京
    • Year and Date
      2009-09-04
    • Related Report
      2009 Annual Research Report
  • [Presentation] Highly Sensitive Analysis of Sugar Chains in Capillary Electrophoresis2009

    • Author(s)
      川井隆之
    • Organizer
      HPLC2009
    • Place of Presentation
      Dresden, Germany
    • Year and Date
      2009-06-30
    • Related Report
      2009 Annual Research Report
  • [Presentation] キャピラリー電気泳動における糖鎖分析の高感度化(3)2009

    • Author(s)
      川井隆之
    • Organizer
      第16回クロマトグラフィーシンポジウム
    • Place of Presentation
      長崎
    • Year and Date
      2009-05-29
    • Related Report
      2009 Annual Research Report
  • [Presentation] キャピラリー電気泳動における糖鎖分析の高感度化(2)2009

    • Author(s)
      川井隆之
    • Organizer
      第70回分析化学討論会
    • Place of Presentation
      和歌山
    • Year and Date
      2009-05-16
    • Related Report
      2009 Annual Research Report

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Published: 2009-04-01   Modified: 2024-03-26  

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