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2003 Fiscal Year Final Research Report Summary

Thermoresponsive chromatography with regulated electrostatic interactions for effective separation of nucleic acids

Research Project

Project/Area Number 13680955
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Biomedical engineering/Biological material science
Research InstitutionTokyo Women's Medical University

Principal Investigator

KIKUCHI Akihiko  Tokyo Women's Medical University, Department of Medicine, Associate Professor, 医学部, 助教授 (40266820)

Project Period (FY) 2001 – 2003
Keywordsthermoresponsive polymers / poly(N-isopropylacrylamide) / cationic groups / chromatography / oligonucleotide / nucleoside base / hydrophobic interaction / electrostatic interaction
Research Abstract

This research focused on the design and characterization of novel thermoresponsive chromatography system in which the surface characteristics can change hydrophilic/hydrophobic properties and, at the same time, surface charge densities by temperature as a sole variant in aqueous milieu. Such surfaces was prepared for separation of bioactive nucleic acids efficiently without use of several chromatography columns working with different separation modes. We have already elucidated the surface property alterations of the cationic thermoresponsive poly(N-isopropylacrylamide) (PIPAAm) modified surfaces with temperature in terms of hydrophobicity changes and surface potential changes. Furthermore, separation of adenosine nucleotides, AMP, ADP, and ATP was successfully achieved with this column matrix. In this fiscal year, separation of adenosine oligonucleotides and thymidine oligonucleotides with repeating number of 2-5, were investigated using cationic, thermoresponsive column. Thermorespon … More sive column matrices were prepared by radical polymerization of IPAAm monomer and 3,3-dimethylaminopropylacrylamide (DMAPAA) on the initiator immobilized silica beads. The sample mixture was injected into polymer-modified silica packed column (4.6mmφ, 150mm length), and elution behavior of these nucleotides were monitored. For nonionic PIPAAm homopolymer column, retention times of the oligonucleotides are almost same regardless of the number of repeating units. In contrast, retention of oligonucleotides is increased with increasing number of repeating units on cationic, thermoresponsive surfaces. Polarity of the nucleotide bases is significantly important to alter oligonucleotide retention on cationic thermoresponsive column matrices. Thus the retention times of the thymidine oligonucleotides are slightly longer than those of the adenosine nucleotides with the same amount of the phosphate groups. The interaction of oligonucleotides and cationic surfaces is multi-point interaction of polyelectrolytes, and thus the retention increased with increasing the number of repeating units. In conclusion, the obtained results suggest that the prepared cationic, thermoresponsive surfaces can be used to separate oligonucleotides through the recognition of charge densities and hydrophobicity of sample molecules. Less

  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] 菊池明彦, 岡野光夫: "水系で生理活性物質を分離するグリーンクロマトグラフィー"バイオマテリアル-生体材料-. 21(2). 133-141 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 菊池明彦: "インテリジェント表面"バイオマテリアル-生体材料-. 21(3). 214-215 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 坂本千賀子, 岡田裕司, 金澤秀子, 菊池明彦, 岡野光夫: "温度応答性クロマトグラフィーによるカテキン類の分離"分析化学. 52(10). 903-906 (2003)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Kikuchi, T.Okano: "Temperature-responsive polymer-modified surfaces for green chromatography"Macromolecular Symposia. 207(1). 217-228 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] C.Sakamoto.Y.Okada.H.Kanazawa, E.Ayano, T.Nishimura, M.Ando, A.Kikuchi, T.Okano: "Temperature- and pH-responsive aminopropyl silica ion-exchange columns grafted with copolymers of N-isopropylacryiamide"Journal of Chromatography A. 1030. 247-253 (2004)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akihiko KIKUCHI, Teruo OKANO: "Green chromatography systems for separation of bioactive compounds in aqueous phase"Biomaterials-SEITAI ZAIRYO-(Journal of Japanese Society for Biomaterials). 21(2). 133-141 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akihiko KIKUCHI: "Intelligent interfaces for tissue engineering"Biomaterials-SEITAI ZAIRYO-(Journal of Japanese Society for Biomaterials). 21(3). 214-215 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chikako SAKAMOTO, Yuji OKADA, Hideko KANAZAWA, Akihiko KIKUCHI, Teruo OKANO: "Separation of catechins by temperature-responsive chromatography"BUNSEKI KAGAKU. 52(10). 903-906 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akihiko KIKUCHI, Teruo OKANO: "Temperature-responsive polymer-modified surfaces for green chromatography"Macromolecular Symposia. 207(1). 217-228 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chikako SAKAMOTO, Yuji OKADA, Hideko KANAZAWA, Eri AYANO, T.NISHIMURA, M.ANDO, Akihiko KIKUCHI, Teruo OKANO: "Temperature-and pH-responsive aminopropyl silica ion-exchange columns grafted with copolymers of N-isopropylacrylamide"Journal of Chromatography A. 1030. 247-253 (2004)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2005-04-19  

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