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Synthesis of polymer-enzyme conjugates for site-specific modification of therapeutic proteins

Research Project

Project/Area Number 18K06602
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
Research InstitutionGihu University of Medical Science (2020)
Gifu Pharmaceutical University (2018-2019)

Principal Investigator

Sasai Yasushi  岐阜医療科学大学, 薬学部, 教授 (60336633)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Keywords高分子-酵素複合体 / バイオコンジュゲーション / 温度応答性高分子 / 原子移動ラジカル重合 / 酵素-高分子複合体 / グラフト共重合体 / 高分子-酵素複合体 / トランスグルタミナーゼ / タンパク質医薬品 / PEG化
Outline of Final Research Achievements

We synthesized a thermoresponsive graft-copolymer consisting of vinylmethylether-maleic acid copolymer(VEMA) backbone and UCST-type thermoresponsive poly(sulfobetaine methacrylate)(PSBMA) side chains. The UCST of the graft copolymers was influenced by the grafting and polymerization degree of the PSBMA side chains. Below the UCST, the graft-copolymers formed micelle-like aggregates with a PSBMA core and a VEMAC outer layer. Trypsin as a model enzyme was successfully immobilized onto the aggregates via a multi-point attachment with VEMA. The resultant trypsin-immobilized particles are then in solution for use above the UCST. Furthermore, by lowering the solution temperature below the UCST, the conjugates can be easily collected from the reaction mixture as precipitates to use again.

Academic Significance and Societal Importance of the Research Achievements

本研究で開発した温度応答性グラフトコポリマーは、酵素の安定性に適した低温環境では親水性のvinylmethylether-maleic acid copolymer(VEMA)を表面とする100-200 nmの高分子ミセル様粒子を形成し、多くの酵素活性に適した37℃付近では均一な溶液状態となる。このような温度に応答する溶解特性を持つグラフトコポリマーで修飾された酵素は、反応効率に優れる均一系での使用が可能であり、また、酵素活性の低下を抑制する低温では不溶性粒子として反応系から回収可能である。本研究成果は高価、希少な有用酵素を有効活用するための酵素機能化法として有益な知見を提供するものである。

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report
  • Research Products

    (8 results)

All 2019 2018

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Presentation (7 results) (of which Int'l Joint Research: 2 results)

  • [Journal Article] Effects of Plasma Surface Treatment on Cell Adhesion to Biocompatible Polymer Brushes2019

    • Author(s)
      Yasushi Sasai, Naoki Doi, Yukinori Yamauchi, Masayuki Kuzuy, Shin-ichi Kondo
    • Journal Title

      Journal of Photopolymer Science and Technology

      Volume: 32 Issue: 3 Pages: 529-533

    • DOI

      10.2494/photopolymer.32.529

    • NAID

      130007744426

    • ISSN
      0914-9244, 1349-6336
    • Year and Date
      2019-06-24
    • Related Report
      2019 Research-status Report
    • Peer Reviewed
  • [Presentation] Self-assembly behavior of graft copolymer with thermoresponsive side chains and its evaluation2019

    • Author(s)
      Yasushi Sasai, Masao Morita, Naoki Doi, Yukinori Yamauchi, Masayuki Kuzuya, Shin-ichi Kondo
    • Organizer
      第68回日本高分子学会年次大会
    • Related Report
      2019 Research-status Report
  • [Presentation] 生体適合性高分子ブラシへの細胞接着におけるプラズマ表面処理の効果2019

    • Author(s)
      笹井泰志, 土井直樹, 山内行玄, 葛谷昌之, 近藤伸一
    • Organizer
      第36 回国際フォトポリマーコンファレンス
    • Related Report
      2019 Research-status Report
  • [Presentation] 酵素固定化担体への展開を指向したセルロースナノファイバーの表面機能化2019

    • Author(s)
      笹井泰志、南田陽日、土井直樹、山内行玄、葛谷昌之、近藤伸一
    • Organizer
      日本薬学会第139年会
    • Related Report
      2018 Research-status Report
  • [Presentation] 温度応答性グラフト共 重合体-トリプシンコンジュゲートの合成とその評価2018

    • Author(s)
      笹井泰志、森田将生、土井直樹、山内行玄、葛谷昌之、近藤伸一
    • Organizer
      第67回高分子学会年次大会
    • Related Report
      2018 Research-status Report
  • [Presentation] メタクリル酸系双性イオン高分子のプラズマ誘起反応2018

    • Author(s)
      笹井泰志、中牟田晧平、土井直樹、山内行玄、葛谷昌之、近藤伸一
    • Organizer
      第35回国際フォトポリマーカンファレンス
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research
  • [Presentation] 温度応答性高分子ミセル様粒子の調製とその酵素固定化担体としての応用2018

    • Author(s)
      森田将生、笹井泰志、土井直樹、山内行玄、葛谷昌之、近藤伸一
    • Organizer
      第64回日本薬学会東海支部 総会・大会
    • Related Report
      2018 Research-status Report
  • [Presentation] A strategy for synthesizing polymer-trypsin conjugates by using UCST-type thermoresponsive system2018

    • Author(s)
      笹井泰志、森田将生、土井直樹、山内行玄、葛谷昌之、近藤伸一
    • Organizer
      The 12th SPSJ International Polymer Conference (IPC2018)
    • Related Report
      2018 Research-status Report
    • Int'l Joint Research

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Published: 2018-04-23   Modified: 2022-01-27  

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