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Development of the Methodology for Dissociation Kinetic Analysis of Biomolecular Complexes with Capillary Electrophotetic Reactor

Research Project

Project/Area Number 15350039
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Analytical chemistry
Research InstitutionTohoku University

Principal Investigator

TAKAHASHI Toru (2004)  Tohoku University, Graduate school of environmental studies, Research associate, 大学院・環境科学研究科, 助手 (30361166)

星野 仁 (2003)  東北大学, 大学院・環境科学研究科, 教授 (20124620)

Co-Investigator(Kenkyū-buntansha) IKI Nobuhiko  Tohoku University, Graduate school of environmental studies, Associate professor, 大学院・環境科学研究科, 助教授 (50282108)
SAITOH Shingo  Kitami Institute of Technology, Graduate school of engineering, Research associate, 大学院・工学研究科, 助手 (60343018)
高橋 透  東北大学, 大学院・環境科学研究科, 助手 (30361166)
Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥13,100,000 (Direct Cost: ¥13,100,000)
Fiscal Year 2004: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 2003: ¥6,300,000 (Direct Cost: ¥6,300,000)
Keywordscapillary electrophoretic reactor / microchip capillary electroresis / microchip capillary electrohoretic reactor / dissoclation kenetic analysis / metal complexes / polyaminocaoboxylic acid / biomolecular complexes / Microchi Caillary Eledrohoretic Reactor / μCER / 解離反応速度論解析法 / Quin-2 / Ce(III)錯体 / ハイスループット / キャピラリー電気泳動流動反応器 / 抗原-抗体 / キャピラリーゾーン電気泳動分離 / 解離反応速度解析
Research Abstract

A microchip capillary electrophoretic reactor (MCER), a practical and powerful tool for dissociation kinetic analysis of metal complexes, has been proposed. In the dissociation kinetic analysis of metal complexes using conventional batch reaction techniques, owing to the difficulty in dividing between reactant (metal complex) and products (metal ion and free ligand), the direct measurement of the dissociation kinetics had not been achieved. On the contrary, MCER revolutionized the measurement of the dissociation rate constant of metal complexes since its accomplishment of the direct measurement. The basic idea of a MCER was provoked by the concentration jump character of the sharp band profiles in microchip capillary electrophoresis (MCE) ; i.e. metal complexes during the MCE separation processes, in which there had been no ligand in the electrophoretic buffer solution, were exposed to overwhelming force to dissociation caused by a steep concentration jump of the ligand. The time depending dissociation reaction profile of metal complex of Ce^<3+> with a polyaminocarboxylic ligand, 8-amino-2-[(2-amino-5-methylphenoxyl)methyl]-6-methoxyquinoline-N,N,N',N',-tetraacetic acid ([CeL]^-) was described using both internal and external standard. The solvolytic dissociation rate constant of [CeL]^-have been determined directly as 6.2 X 10^<-3> s^<-1> at 303 K with MCER.
Capillary electrophoretic reactor (CER) was applied to the determination of dissociation reaction rate constant of bio-molecular complexes. Both an immunocomplex of anti digoxin IgG with digoxin and a complex double stranded DNA with ethidium bromide were chosen as analytical model compounds. Unfortunately, determination of dissociation reaction rate constants could not be achieved with CER in both systems, since their properties of small electrophoretic mobility and lower kinetically inertness.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (5 results)

All 2004

All Journal Article (5 results)

  • [Journal Article] Highly-sensitive simultaneous detection of lanthanide(III) ions as kinetically stable aromatic polyaminocarboxytato complexes bia capillary electrophoresis using resolution enhancement with carbonate ion2004

    • Author(s)
      Shingo Saitoh
    • Journal Title

      Analytical and Bioanalytical Chemistry 378

      Pages: 1644-1647

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Sulfonylcalix[4]arenetetrasulfonate as pre-column chelating reagent for selective determination of aluminum(III), iron(III), and titanium(IV) by ion-pair reversed phase high-performance liquid chromatography with spectrophotometric detection2004

    • Author(s)
      Hiroaki Matsumiya
    • Journal Title

      Talanta 62

      Pages: 337-342

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Highly-selective simultaneous detection of lanthanide(III) ions as kinetically stable aromatic polyaminocarboxylatato complexes via capillary electrophoresis using resolution enhancement with carbonate ion2004

    • Author(s)
      Shingo Saito
    • Journal Title

      Analytical and Bioanalytical Chemistry 378

      Pages: 1644-1647

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Sulfonilcalix[4]arenetetrasulfonate as pre-column chelating reagent for selective determination of aluminum(III), iron(III), and titanium(IV) by ion-pair reversed phase high-performance liquid chromatography with spectrophotometric detection2004

    • Author(s)
      Hioroaki Matsumiya
    • Journal Title

      Talanta 62

      Pages: 337-342

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Highly-selectice simultaneous detection of lanthanide(III) ions as kinetically stable aromatic polyaminocarboxylatato complexes via capillary electrophoresis using resolution enhancement with carbonate ion2004

    • Author(s)
      Shingo Saito
    • Journal Title

      Analytical and Bioanalytical Chemistry 378

      Pages: 1644-1647

    • Related Report
      2004 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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