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Control of liquid-liquid phase separation of protein by small additives

Research Project

Project/Area Number 18H02383
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Review Section Basic Section 43020:Structural biochemistry-related
Research InstitutionUniversity of Tsukuba

Principal Investigator

Shiraki Kentaro  筑波大学, 数理物質系, 教授 (90334797)

Co-Investigator(Kenkyū-buntansha) 冨田 峻介  国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (50726817)
加納 英明  九州大学, 大学院理学研究院, 教授 (70334240)
Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥17,810,000 (Direct Cost: ¥13,700,000、Indirect Cost: ¥4,110,000)
Fiscal Year 2020: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2019: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2018: ¥11,050,000 (Direct Cost: ¥8,500,000、Indirect Cost: ¥2,550,000)
Keywordsタンパク質 / 高濃度 / 凝集 / 液-液相分離 / 低分子 / コンデンセート / 酵素 / 液滴 / ドロプレット / CARS顕微鏡 / タンパク質凝集 / 濃縮
Outline of Final Research Achievements

Proteins are prone to form aggregates and liquid droplets. We aimed to clarify the mechanism of stabilization of the solution state of high-concentration proteins and to establish a method to control the solution states. It was found that when different proteins derived from egg white were mixed in a low ionic strength solution, they formed a liquid-liquid phase separated state, but matured with time to become co-aggregated. Alcohols and polyion complexes were found to be effective in stabilizing the phase-separated droplets, but this was different from the stabilization of aggregates. These studies have enhanced our understanding of the solution states of highly concentrated proteins.

Academic Significance and Societal Importance of the Research Achievements

高濃度のタンパク質の溶液状態の制御は、食品や医薬品の開発のために重要な知見を与える。本研究は集合した状態のタンパク質を低分子で制御する方法の確立を目指した。卵白由来のタンパク質の凝集や液-液相分離の低分子による制御は、食品産業への応用が期待できる。ポリアミノ酸による抗体の安定化は、バイオ医薬品への応用が期待できる。タンパク質の凝集体を選択的に除去するための仕組みとして水性二相溶液が利用できることは、課題となっている目に見えない凝集体除去の方法の開発に役立つだろう。

Report

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

    (17 results)

All 2021 2020 2019 2018

All Journal Article (9 results) (of which Peer Reviewed: 6 results) Presentation (6 results) (of which Invited: 2 results) Book (2 results)

  • [Journal Article] Glass-like protein condensate for the long-term storage of proteins2021

    • Author(s)
      Nakauchi Yoshitaka、Nishinami Suguru、Shiraki Kentaro
    • Journal Title

      International Journal of Biological Macromolecules

      Volume: 182 Pages: 162-167

    • DOI

      10.1016/j.ijbiomac.2021.04.017

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Aggregation of Hen Egg White Proteins with Additives During Agitation2021

    • Author(s)
      Hong Taehun、Iwashita Kazuki、Han Jeungmin、Nishinami Suguru、Handa Akihiro、Shiraki Kentaro
    • Journal Title

      LWT

      Volume: Mar 27 Pages: 111378-111378

    • DOI

      10.1016/j.lwt.2021.111378

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Salt-containing aqueous two-phase system shows predictable partition of proteins with surface amino acids residues2019

    • Author(s)
      Shibata Chika、Iwashita Kazuki、Shiraki Kentaro
    • Journal Title

      International Journal of Biological Macromolecules

      Volume: 133 Pages: 1182-1186

    • DOI

      10.1016/j.ijbiomac.2019.04.185

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Selective separation method of aggregates from IgG solution by aqueous two-phase system2019

    • Author(s)
      Shibata Chika、Iwashita Kazuki、Shiraki Kentaro
    • Journal Title

      Protein Expression and Purification

      Volume: 161 Pages: 57-62

    • DOI

      10.1016/j.pep.2019.05.002

    • Related Report
      2019 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Control of Aggregation, Coaggregation, and Liquid Droplet of Proteins Using Small Additives2019

    • Author(s)
      Iwashita Kazuki、Mimura Masahiro、Shiraki Kentaro
    • Journal Title

      Current Pharmaceutical Biotechnology

      Volume: 19 Issue: 12 Pages: 946-955

    • DOI

      10.2174/1389201020666181204113054

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Effect of additives on liquid droplet of protein?polyelectrolyte complex for high-concentration formulations2019

    • Author(s)
      Mimura Masahiro、Tsumura Keisuke、Matsuda Ayumi、Akatsuka Naoki、Shiraki Kentaro
    • Journal Title

      The Journal of Chemical Physics

      Volume: 150 Issue: 6 Pages: 064903-064903

    • DOI

      10.1063/1.5063378

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Non-chromatographic purification of Teriparatide with a pH-responsive CspB tag2019

    • Author(s)
      Nonaka Takahiro、Tsurui Noriko、Mannen Teruhisa、Kikuchi Yoshimi、Shiraki Kentaro
    • Journal Title

      Protein Expression and Purification

      Volume: 155 Pages: 66-71

    • DOI

      10.1016/j.pep.2018.11.008

    • Related Report
      2018 Annual Research Report
  • [Journal Article] Coacervates and coaggregates: Liquid?liquid and liquid?solid phase transitions by native and unfolded protein complexes2018

    • Author(s)
      Iwashita Kazuki、Handa Akihiro、Shiraki Kentaro
    • Journal Title

      International Journal of Biological Macromolecules

      Volume: 120 Pages: 10-18

    • DOI

      10.1016/j.ijbiomac.2018.08.063

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Liquid Droplet of Protein-Polyelectrolyte Complex for High-Concentration Formulations2018

    • Author(s)
      Matsuda Ayumi、Mimura Masahiro、Maruyama Takuya、Kurinomaru Takaaki、Shiuhei Mieda、Shiraki Kentaro
    • Journal Title

      Journal of Pharmaceutical Sciences

      Volume: 107 Issue: 10 Pages: 2713-2719

    • DOI

      10.1016/j.xphs.2018.06.021

    • Related Report
      2018 Annual Research Report
  • [Presentation] 液-液相分離によるIgGの濃縮法2019

    • Author(s)
      三村真大、津村圭亮、栗田僚二、白木賢太郎
    • Organizer
      第19回日本蛋白質科学会大会 第71回細胞生物物理学会大会 合同年次大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Liquid-liquid phase separation and fibrillization of yeast prion Sup35 NM domain2019

    • Author(s)
      Suguru Nishinami, Yumiko Ohhashi, Kentaro Shiraki
    • Organizer
      第19回日本蛋白質科学会大会 第71回細胞生物物理学会大会 合同年次大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] 体膜のない細胞内小器官を模倣した PLL/ATP 液滴の研究2019

    • Author(s)
      浦朋人、白木賢太郎
    • Organizer
      第19回日本蛋白質科学会大会 第71回細胞生物物理学会大会 合同年次大会
    • Related Report
      2019 Annual Research Report
  • [Presentation] Effects of co-solutes on liquid-liquid phase separation and fibrillization of yeast prion Sup32019

    • Author(s)
      Suguru Nishinami, Yumiko Ohhashi, Kentaro Shiraki
    • Organizer
      The 57th Annual Meeting of The Biophysical Society of Japan
    • Related Report
      2019 Annual Research Report
  • [Presentation] 相分離生物学2018

    • Author(s)
      白木賢太郎
    • Organizer
      第1回LLPS研究会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] タンパク質の凝集・共凝集・相分離のメカニズムと産業応用2018

    • Author(s)
      白木賢太郎
    • Organizer
      第80回酵素工学研究会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Book] 相分離生物学の全貌(現代化学増刊46)2020

    • Author(s)
      白木 賢太郎
    • Total Pages
      400
    • Publisher
      東京化学同人
    • ISBN
      9784807913466
    • Related Report
      2020 Annual Research Report
  • [Book] 相分離生物学2019

    • Author(s)
      白木 賢太郎
    • Total Pages
      176
    • Publisher
      東京化学同人
    • ISBN
      9784807909650
    • Related Report
      2019 Annual Research Report

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

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