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In situ photopolymerization of polyacryrlamide gels for specific entrapment and analysis of carbohydrates using microchip electrophoresis

Research Project

Project/Area Number 15K18852
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Physical pharmacy
Research InstitutionKindai University

Principal Investigator

YAMAMOTO Sachio  近畿大学, 薬学部, 助教 (10707954)

Project Period (FY) 2015-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsマイクロチップ電気泳動 / 糖鎖 / オンライン濃縮 / 酵素消化 / オンライン標識 / オンチップ法 / 糖タンパク質糖鎖 / 糖鎖調製
Outline of Final Research Achievements

For the fast releasing of N-linked glycans from glycoproteins, a simple and efficient method has been developed by fabricating trypsin and peptide-N4-(N-acetyl-β-glucosaminyl)asparagine amidase (PNGase F)-impregnated polyacrylamide gel onto a commercial pipette tip, respectively. The fixed enzymes maintained their activities on the polyacrylamide gel and the reaction was completed by a few times pipetting operation.Also, a simple and efficient method has been developed to fabricate sample concentrate and derivative on a channel of a commercial polymethylmethacrylate-made microchip The availability of ionic acrylamide gel was demonstrated for the sensitive analysis of some FITC-amino acids under various buffer systems at several different pH. Then, the multi channel microchip was developed for combination of various functions of acrylamide gel, and achieves high-throughput glycans analysis.

Report

(3 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • Research Products

    (20 results)

All 2017 2016 2015

All Journal Article (7 results) (of which Peer Reviewed: 7 results,  Acknowledgement Compliant: 5 results,  Open Access: 1 results) Presentation (13 results) (of which Int'l Joint Research: 1 results,  Invited: 1 results)

  • [Journal Article] Online Cleanup Liquid Chromatography for the Analysis of Glycoprotein-Derived Oligosaccharides Labeled with 7-Amino-4-methylcoumarin2017

    • Author(s)
      Yoshie NAGATOMO, Shinichi HASHIMOTO, Yuka KISHIMOTO, Takao HAYAKAWA, Sachio YAMAMOTO, Mitsuhiro KINOSHITA, Shigeo SUZUKI
    • Journal Title

      CHROMATOGRAPHY

      Volume: 38 Issue: 1 Pages: 23-30

    • DOI

      10.15583/jpchrom.2016.016

    • NAID

      130005439673

    • ISSN
      1342-8284, 1348-3315
    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Plug-plug Kinetic Capillary Electrophoresis for In-capillary Exoglycosidase Digestion as a Profiling Tool for the Analysis of Glycoprotein Glycans2017

    • Author(s)
      Maki Yamagami, Yurie Matsui, Takao Hayakawa, Sachio Yamamoto, Mitsuhiro Kinoshita, Shigeo Suzuki
    • Journal Title

      Journal of Chromatography A

      Volume: 1496 Pages: 157-162

    • DOI

      10.1016/j.chroma.2017.03.019

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Current landscape of protein glycosylation analysis and recentprogress toward a novel paradigm of glycoscience research2016

    • Author(s)
      Sachio Yamamoto, Mitsuhiro Kinoshita, Shigeo Suzuki
    • Journal Title

      Journal of Pharmaceutical and Biomedical Analysis

      Volume: 130 Pages: 273-300

    • DOI

      10.1016/j.jpba.2016.07.015

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Capillary electrochromatography using monoamine- andtriamine-bonded silica nanoparticles as pseudostationary phases2016

    • Author(s)
      Yuto Takeda, Yuka Hayashi, Naonori Utamura, Chise Takamoto, Mitsuhiro Kinoshita,Sachio Yamamoto, Takao Hayakawa, Shigeo Suzuki
    • Journal Title

      Journal of Chromatography A

      Volume: 1427 Pages: 170-176

    • DOI

      10.1016/j.chroma.2015.11.085

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] A rapid and highly sensitive microchip electrophoresis of mono- and mucin-type oligosaccharides labeled with 7-amino-4-methylcoumarin2015

    • Author(s)
      Sachio Yamamoto, Eriko Nagai, Yuki Asada, Mitsuhiro Kinoshita, Shigeo Suzuki
    • Journal Title

      Analytical and Bioanalytical Chemistry

      Volume: 407 Issue: 5 Pages: 1499-1503

    • DOI

      10.1007/s00216-014-8363-x

    • Related Report
      2015 Research-status Report
    • Peer Reviewed
  • [Journal Article] Synthesis of Quaternary Ammonium Derivatives of Cellulose as the Coating Reagents for Capillary Electrophoresis2015

    • Author(s)
      Sachio Yamamoto, Tomoyuki Iwata, Keiji Nishiwaki, Mitsuhiro Kinoshita, Shigeo Suzuki
    • Journal Title

      CHROMATOGRAPHY

      Volume: 36 Issue: 3 Pages: 93-98

    • DOI

      10.15583/jpchrom.2015.027

    • NAID

      130005118200

    • ISSN
      1342-8284, 1348-3315
    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Chiral separation of D/L-aldoses by micellar electrokinetic chromatography using a chiral derivatization reagent and a phenylboronic acid complex2015

    • Author(s)
      Sachio Yamamoto, Yoko Tamata, Kaori Sejima, Mitsuhiro Kinoshita, Shigeo Suzuki
    • Journal Title

      Analytical and Bioanalytical Chemistry

      Volume: 407 Issue: 20 Pages: 6201-6206

    • DOI

      10.1007/s00216-015-8802-3

    • Related Report
      2015 Research-status Report
    • Peer Reviewed
  • [Presentation] 酵素固定化アクリルアミドゲルチップを用いる糖タンパク質糖鎖調製法の開発2017

    • Author(s)
      山本佐知雄、植田麻希、中村朋子、木下充弘、鈴木茂生
    • Organizer
      薬学会第137年会
    • Place of Presentation
      仙台国際センター(宮城)
    • Year and Date
      2017-03-24
    • Related Report
      2016 Annual Research Report
  • [Presentation] DCpak PTZカラムを用いた8-aminopyrene-1,3,6- trisulfonic acid (APTS)標識化糖タンパク質由来糖鎖のHPLC分析2016

    • Author(s)
      山本 佐知雄,木下充弘,池上亨,西原啓二,鈴木茂生
    • Organizer
      第27回クロマトグラフィー科学会議
    • Place of Presentation
      慶應義塾大学薬学部[東京]
    • Year and Date
      2016-11-16
    • Related Report
      2016 Annual Research Report
  • [Presentation] フェニルボロン酸添加MEKCによる(S)-(+)-4-(N,N-Dimethylaminosulfonyl)-7-(3-aminopyrrolidin-1-yl)-2,1,3-benzoxadiazol標識化D-,L-糖の光学分離2016

    • Author(s)
      山本佐知雄、田又洋子、瀬島佳保里、木下充弘、鈴木茂生
    • Organizer
      SCE2016
    • Place of Presentation
      徳島大学(徳島)
    • Year and Date
      2016-11-09
    • Related Report
      2016 Annual Research Report
  • [Presentation] 光重合性アクリルアミドゲルを用いるリン酸化化合物のオンライン濃縮マイクロ チップ電気泳動法の開発2016

    • Author(s)
      山本佐知雄,姫野美幸,小林正弥,赤松美紀,木下充弘,鈴木茂生
    • Organizer
      SCE2016
    • Place of Presentation
      徳島大学(徳島)
    • Year and Date
      2016-11-09
    • Related Report
      2016 Annual Research Report
  • [Presentation] In situ photopolymerization of polyacryrlamide gel for specific entrapment and analysis of a phosphate compounds using microchip electrophoresis2016

    • Author(s)
      Sachio Yamamoto, Miyuki Himeno, Masaya Kobayashi, Mitsuhiro Kinoshita, Shigeo Suzuki
    • Organizer
      12th International Conference on Protein Phosphatase
    • Place of Presentation
      Kindai university (Japan)
    • Year and Date
      2016-10-27
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 酵素固定化アクリルアミドゲルチップ を用いる糖タンパク質糖鎖調製法の開発2016

    • Author(s)
      山本 佐知雄、植田 麻希、中村 朋子、木下 充弘、鈴木 茂生
    • Organizer
      日本分析化学会第65年会
    • Place of Presentation
      北海道大学(北海道)
    • Year and Date
      2016-09-14
    • Related Report
      2016 Annual Research Report
  • [Presentation] 光硬化性アクリルアミドゲルのピンポ イント作製技術を用いる糖鎖の構造解析法の開発2016

    • Author(s)
      山本 佐知雄、大西 翔太、鈴木 翔、木下 充弘、鈴木 茂生
    • Organizer
      第76回分析化学討論会
    • Place of Presentation
      岐阜薬科大学(岐阜)
    • Year and Date
      2016-05-28
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 光硬化性アクリルアミドゲルのピンポイント作製技術を用いる糖鎖の構造解析法の開発2016

    • Author(s)
      山本 佐知雄,大西 翔太,鈴木 翔,木下 充弘,鈴木 茂生
    • Organizer
      日本分析化学会 第76回分析化学討論会
    • Place of Presentation
      岐阜大学(岐阜県)
    • Year and Date
      2016-05-28
    • Related Report
      2015 Research-status Report
  • [Presentation] 光重合性アクリルアミドゲルを用いるリン酸化化合物の オンライン濃縮マイクロチップ電気泳動法の開発2016

    • Author(s)
      山本佐知雄,小林正弥, 丸井戸翔,姫野 美幸 , 木下充弘,鈴木茂生
    • Organizer
      日本薬学会第136年会
    • Place of Presentation
      パシフィコ横浜(神奈川県)
    • Year and Date
      2016-03-26
    • Related Report
      2015 Research-status Report
  • [Presentation] 光重合性アクリルアミドゲルを用いるリン酸化化合物のオンライン濃縮マイクロチップ電気泳動法の開発2016

    • Author(s)
      山本佐知雄,姫野美幸,小林正弥,赤松美紀,木下充弘,鈴木茂生
    • Organizer
      新アミノ酸分析研究会第6回学術講演会
    • Place of Presentation
      東京大学武田先端知ビル(東京)
    • Related Report
      2016 Annual Research Report
  • [Presentation] 光硬化性アクリルアミドゲルによるオンライン濃縮マイクロチップ電気泳動を用いるシグナル伝達経路におけるリン酸化ペプチドの発現量の解析2015

    • Author(s)
      山本佐知雄,丸井戸翔,赤松美紀, 木下充弘,鈴木茂生
    • Organizer
      化学とマイクロ・ナノシステム学会 第32回研究会(32nd CHEMINAS)
    • Place of Presentation
      北九州国際会議場(福岡県)
    • Year and Date
      2015-11-26
    • Related Report
      2015 Research-status Report
  • [Presentation] 酵素固定化アクリルアミドゲルチップを用いる糖タンパ質鎖調製法の開発2015

    • Author(s)
      山本 佐知雄,植田 麻希,葛西 優貴,木下 充弘,鈴木 茂生
    • Organizer
      第 26 回クロマトグラフィー科学会議
    • Place of Presentation
      九州大学(福岡県)
    • Year and Date
      2015-11-11
    • Related Report
      2015 Research-status Report
  • [Presentation] 酵素固定化アクリルアミドゲルチップを用いる糖タンパク質糖鎖調製法の開発2015

    • Author(s)
      山本 佐知雄,植田 麻希,葛西 優貴,木下 充弘,鈴木 茂生
    • Organizer
      第28回バイオメディカル分析科学シンポジウム (BMAS2015)
    • Place of Presentation
      長崎大学(長崎県)
    • Year and Date
      2015-08-21
    • Related Report
      2015 Research-status Report

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Published: 2015-04-16   Modified: 2020-01-20  

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