Development of analysis technique for accelerating protein aggregation and crystallization under applied electric fields and the clarification of the mechanism

• Project/Area Number: 26390049
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied materials
• Research Institution: Fukushima National College of Technology (2016); Ibaraki National College of Technology (2014-2015)
• Principal Investigator: Wakamatsu Takashi 福島工業高等専門学校, 電気工学科, 教授 (80220838)
• Co-Investigator(Kenkyū-buntansha): 千葉 薫 茨城工業高等専門学校, 自然科学科, 准教授 (50415775)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2014: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: タンパク質 / 凝集体 / 結晶化 / 光散乱 / 電場刺激 / フラクタル / 微粒子

Outline of Final Research Achievements

We have developed a new analysis apparatus with the ability to in situ measure forward scattering light at small angle for crystallizing protein solutions under a low applied voltage.
We tracked protein lysozyme aggregation in the pre-crystalline solutions using the developed apparatus. An enhancement of forward scattering intensities was observed for lysozyme solutions with a low-concentration salt under electric fields, which were under not crystallizing condition without applied voltages. We have found that the application of electric fields provides an effect of accelerating a formation of fractal aggregates on the pre-crystalline step.

Research Products

• [Journal Article] Low Applied Voltage Effects on Thaumatin Protein Crystallization (2016)
  • Author(s): Takashi Wakamatsu
  • Journal Title: Transactions of the Materials Research Society of Japan Volume: 41 Issue: 1 Pages: 13-15
  • DOI: 10.14723/tmrsj.41.13
  • NAID: 130005126127
  • ISSN: 1382-3469, 2188-1650
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Method and apparatus for characterization of electric field-induced aggregation in pre-crystalline protein solutions (2015)
  • Author(s): Takashi Wakamatsu
  • Journal Title: Review of Scientific Instruments Volume: 86 Issue: 1
  • DOI: 10.1063/1.4906328
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Forward-Light-Scattering Characterization of Pre-Crystalline Aggregates in Crystallizing Lysozyme Solutions (2014)
  • Author(s): Takashi Wakamatsu
  • Journal Title: American Journal of Analytical Chemistry Volume: 5 Issue: 09 Pages: 581-588
  • DOI: 10.4236/ajac.2014.59065
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 前方光散乱瞬時測定による電場印加におけるタンパク質凝集化の観察 (2017)
  • Author(s): 若松 孝
  • Organizer: 第64回応用物理学会学術講演会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2017-03-14
• [Presentation] Characterization of Electric Field-Induced Aggregation of Lysozyme using In Situ Forward-Light-Scattering Measurement Technique （前方光散乱瞬時計測法によるリゾチームの電場誘起凝集化の評価） (2016)
  • Author(s): 若松 孝
  • Organizer: The 26th Annual Meeting of MRS-Japan 2016 （第26回日本MRS年次大会）
  • Place of Presentation: 横浜情報文化センター（横浜市）
  • Year and Date: 2016-12-19
• [Presentation] 前方散乱光瞬時計測によるリゾチーム凝集・結晶化の評価 (2016)
  • Author(s): 若松 孝
  • Organizer: 第77回応用物理学会学術講演会
  • Place of Presentation: 朱鷺メッセ（新潟市）
  • Year and Date: 2016-09-13
• [Presentation] 前方光散乱法による結晶化前段階形成のリゾチーム凝集体のキャラクタリゼーション (2015)
  • Author(s): 若松 孝
  • Organizer: 第25回日本MRS年次大会
  • Place of Presentation: 神奈川県民ホール(横浜)
  • Year and Date: 2015-12-08
• [Presentation] Effects of Applied Electric Field on Protein Crystallization （電場印加によるタンパク質の結晶化効果） (2014)
  • Author(s): Takashi Wakamatsu （若松 孝）
  • Organizer: The 24th Annual Meeting of MRS-Japan 2014 （第24回日本ＭＲＳ年次大会）
  • Place of Presentation: Yokohama Media & Communications Center
  • Year and Date: 2014-12-10 – 2014-12-12
• [Presentation] 電場印加によるタンパク質凝集・結晶化の促進効果 (2014)
  • Author(s): 若松孝
  • Organizer: 平成26年度電子情報通信学会信越支部大会
  • Place of Presentation: 信州大学長野キャンパス
  • Year and Date: 2014-10-04
• [Presentation] 前方光散乱法による塩のリゾチーム結晶化作用の評価 (2014)
  • Author(s): 若松孝
  • Organizer: 第14回日本蛋白質科学会年会
  • Place of Presentation: 横浜市横浜産貿ホール
  • Year and Date: 2014-06-25 – 2014-06-27
• [Remarks] 第16回バイオ・ライフサイエンス研究展（BIO tech 2017） アカデミックフォーラム
  • URL: http://june2017.tems-system.com/exhiSearch/BIOForum/jp/Workshop
• [Remarks] JST新技術説明会（高専-材料分野)
  • URL: https://shingi.jst.go.jp/list/kosen-k/2016_kosen-k.html
• [Remarks] イノベーション・ジャパン2016 大学見本市
  • URL: http://www.jst.go.jp/tt/fair/ij2016/exhibitor/js20160232.html
• [Remarks] 茨城高専地域共同テクノセンター 教員研究紹介
  • URL: http://www.ibaraki-ct.ac.jp/?page_id=3603/
• [Patent(Industrial Property Rights)] PCT国際出願 (2016)
  • Inventor(s): 若松 孝
  • Industrial Property Rights Holder: 国立高等専門学校機構
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-10-03
  • Overseas
• [Patent(Industrial Property Rights)] ①結晶化分析装置及び結晶化分析方法 (2015)
  • Inventor(s): 国立高等専門学校機構
  • Industrial Property Rights Holder: 国立高等専門学校機構
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2015-198366
  • Filing Date: 2015-10-06