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Development of Purification Process of Plasmid DNA for Gene Therapy by Introducing Affinity Membrane Filtration

Research Project

Project/Area Number 25420800
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Properties in chemical engineering process/Transfer operation/Unit operation
Research InstitutionNagoya University

Principal Investigator

KATAGIRI Nobuyuki  名古屋大学, 工学(系)研究科(研究院), 助教 (00345919)

Co-Investigator(Kenkyū-buntansha) IRITANI Eiji  名古屋大学, 大学院工学研究科, 教授 (60144119)
Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
KeywordsプラスミドDNA / 精製 / アフィニティ / 膜濾過 / リガンド / 吸着 / 脱着 / 菌体破砕
Outline of Final Research Achievements

The development of a purification process of medical products purity plasmid DNA by affinity membrane filtration was examined. From the results of particle size distribution, photomicrograph, membrane filtration of slurry, and electrophoretic analysis of nucleic acids, it was found that the introduction of electroporation was useful for elution of plasmid DNA from bacterial cells. The iron oxide particle was selected as a ligand for two-stage affinity microfiltration of plasmid DNA. In the first stage microfiltration, the experiment was conducted under the condition of pH 5, and the cake of iron oxide particle with plasmid DNA bound was obtained and the impurities were removed. Next, water (pH 9 10 and 12) permeations through the cake were performed to desorb the bound plasmid DNA. It is revealed that the degree of purification of nucleic acid is high from the analysis of the recovery solution, the usefulness of this method has been confirmed.

Report

(4 results)
  • 2015 Annual Research Report   Final Research Report ( PDF )
  • 2014 Research-status Report
  • 2013 Research-status Report
  • Research Products

    (51 results)

All 2016 2015 2014 2013 Other

All Int'l Joint Research (1 results) Journal Article (12 results) (of which Int'l Joint Research: 2 results,  Peer Reviewed: 12 results,  Acknowledgement Compliant: 9 results) Presentation (32 results) (of which Int'l Joint Research: 2 results,  Invited: 2 results) Book (2 results) Remarks (4 results)

  • [Int'l Joint Research] Tamkang University(Taiwan)

    • Related Report
      2015 Annual Research Report
  • [Journal Article] Developments of Blocking Filtration Model in Membrane Filtration2016

    • Author(s)
      Iritani, E. and N. Katagiri
    • Journal Title

      KONA Powder and Particle Journal

      Volume: 33 Issue: 0 Pages: 179-202

    • DOI

      10.14356/kona.2016024

    • NAID

      130005129079

    • ISSN
      0288-4534, 2187-5537
    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Evaluation of Compression-Permeability Characteristics of Microbial Cake Based on Microfiltration Data2016

    • Author(s)
      Katagiri, N., H. Kawahara, Y. Arai and E. Iritani
    • Journal Title

      Sep. Sci. Technol.

      Volume: 51 Issue: 5 Pages: 845-852

    • DOI

      10.1080/01496395.2015.1119849

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Nanocolloid Cake Properties Determined from Step-Up Pressure Filtration with Single-Stage Reduction in Filtration Area2015

    • Author(s)
      Iritani, E., N. Katagiri, R. Nakajima, K.J. Hwang and T.W. Cheng
    • Journal Title

      AIChE J.

      Volume: 61 Issue: 12 Pages: 4426-4436

    • DOI

      10.1002/aic.14967

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Solid-Liquid Separation Properties in Centrifugal Sedimentation of Bidisperse Colloidal Suspension2015

    • Author(s)
      Cao D.Q., E. Iritani and N. Katagiri
    • Journal Title

      J. Chem. Eng. Japan

      Volume: 48 Issue: 7 Pages: 556-563

    • DOI

      10.1252/jcej.14we219

    • NAID

      130005088129

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Constant-Rate and Constant-Pressure Microfiltration Behaviors Involving Particle Permeation and Cake Growth for Binary Dilute Colloidal Mixtures with Different Sizes2015

    • Author(s)
      入谷 英司, 片桐 誠之, 石川義人
    • Journal Title

      KAGAKU KOGAKU RONBUNSHU

      Volume: 41 Issue: 5 Pages: 309-316

    • DOI

      10.1252/kakoronbunshu.41.309

    • NAID

      130005099439

    • ISSN
      0386-216X, 1349-9203
    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Membrane Pore Blocking during Cake Formation in Constant Pressure and Constant Flux Dead-End Microfiltration of Very Dilute Colloids2015

    • Author(s)
      Iritani, E., N. Katagiri, T. Takenaka and Y. Yamashita
    • Journal Title

      Chem. Eng. Sci.

      Volume: 122 Pages: 465-473

    • DOI

      10.1016/j.ces.2014.09.052

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Synergy Effect of Ultrasonication and Salt Addition on Settling Behaviors of Activated Sludge2015

    • Author(s)
      Iritani, E., M. Nishikawa, N. Katagiri and K. Kawasaki
    • Journal Title

      Sep. Purif. Technol.

      Volume: 144 Pages: 177-185

    • DOI

      10.1016/j.seppur.2015.02.028

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Cake Properties of Nanocolloid Evaluated by Variable Pressure Filtration Associated with Reduction in Cake Surface Area2014

    • Author(s)
      Iritani, E., N. Katagiri, R. Nakajima, K.J. Hwang and T.W. Cheng
    • Journal Title

      AIChE J.

      Volume: 60 Issue: 11 Pages: 3869-3877

    • DOI

      10.1002/aic.14601

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] High-Level Deliquoring of Activated Sludge by Ultrahigh- Pressure Expression Combined with Flocculation2014

    • Author(s)
      Iritani, E., N. Katagiri, T. Washizu and K.J. Hwang
    • Journal Title

      Chem. Eng. Sci.

      Volume: 112 Pages: 1-9

    • DOI

      10.1016/j.ces.2014.03.008

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] Combined Effect of Reversible Flocculation and Ultrahigh Pressure in High-Level Deliquoring Attained by Expression of Activated Sludge2014

    • Author(s)
      入谷 英司, 片桐 誠之, 鷲津 拓也, 黄 國楨, 鄭 東文
    • Journal Title

      KAGAKU KOGAKU RONBUNSHU

      Volume: 40 Issue: 5 Pages: 396-403

    • DOI

      10.1252/kakoronbunshu.40.396

    • NAID

      130004691078

    • ISSN
      0386-216X, 1349-9203
    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Determination of Cake Properties in Ultrafiltration of Nano-colloids Based on Single Step-up Pressure Filtration Test2014

    • Author(s)
      Iritani, E., N. Katagiri, M. Tsukamoto and K.J. Hwang
    • Journal Title

      AIChE J.

      Volume: 60 Issue: 1 Pages: 289-299

    • DOI

      10.1002/aic.14262

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Cake Formation and Particle Rejection in Microfiltration of Binary Mixtures of Particles with Two Different Sizes2014

    • Author(s)
      Iritani, E., N. Katagiri, Y. Ishikawa and D.Q. Cao
    • Journal Title

      Sep. Purif. Technol.

      Volume: 123 Pages: 214-220

    • DOI

      10.1016/j.seppur.2013.12.033

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Presentation] 酵母懸濁液の膜濾過における細胞破砕処理の影響2016

    • Author(s)
      片桐誠之、藤井岳、入谷英司
    • Organizer
      化学工学会第81年会
    • Place of Presentation
      関西大学(大阪府)
    • Year and Date
      2016-03-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] 微生物代謝物の膜ファウリング特性に及ぼす多糖とタンパク質の相互作用の影響2016

    • Author(s)
      片桐誠之、笹川崇光、入谷英司
    • Organizer
      化学工学会第81年会
    • Place of Presentation
      関西大学(大阪府)
    • Year and Date
      2016-03-13
    • Related Report
      2015 Annual Research Report
  • [Presentation] Decolorization of Persistent Dye Wastewater by Membrane Reactor Using Enzyme-Containing Culture of White-Rot Fungus2015

    • Author(s)
      Katagiri, N., M. Takatsu and E. Iritani
    • Organizer
      Filtration and Separation Symposium 2015
    • Place of Presentation
      Tokyo (Japan)
    • Year and Date
      2015-11-25
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Decolorization of Dyehouse Effluent by Enzymatic Membrane Reactor Using Cultural Supernatant of White-Rot Fungus2015

    • Author(s)
      Katagiri, N., M. Takatsu and E. Iritani
    • Organizer
      10th European Congress of Chemical Engineering (ECCE10)
    • Place of Presentation
      Nice (France)
    • Year and Date
      2015-09-27
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] プラスミドDNAのアフィニティ膜濾過・精製における菌体からの溶出法の検討2015

    • Author(s)
      香西真仁、片桐誠之、入谷英司
    • Organizer
      化学工学会第47回秋季大会
    • Place of Presentation
      北海道大学(北海道)
    • Year and Date
      2015-09-09
    • Related Report
      2015 Annual Research Report
  • [Presentation] 微生物代謝物の精密濾過特性に及ぼす多糖とタンパク質の相互作用の影響2015

    • Author(s)
      笹川崇光、片桐誠之、入谷英司
    • Organizer
      化学工学会第47回秋季大会
    • Place of Presentation
      北海道大学(北海道)
    • Year and Date
      2015-09-09
    • Related Report
      2015 Annual Research Report
  • [Presentation] 高圧による微生物細胞の破壊が定速濾過挙動に及ぼす影響2015

    • Author(s)
      桑島侑也、片桐誠之、入谷英司
    • Organizer
      化学工学会第47回秋季大会
    • Place of Presentation
      北海道大学(北海道)
    • Year and Date
      2015-09-09
    • Related Report
      2015 Annual Research Report
  • [Presentation] 細胞模倣ベシクルの膜濾過特性の評価2015

    • Author(s)
      澤田剛、片桐誠之、入谷英司
    • Organizer
      化学工学会第47回秋季大会
    • Place of Presentation
      北海道大学(北海道)
    • Year and Date
      2015-09-09
    • Related Report
      2015 Annual Research Report
  • [Presentation] 活性汚泥法におけるフロック形成と分離特性2015

    • Author(s)
      片桐誠之
    • Organizer
      土・水・生命環境とコロイド界面現象2015
    • Place of Presentation
      筑波大学(茨城県)
    • Year and Date
      2015-07-31
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 嫌気活性汚泥の有機酸産生と膜濾過特性に及ぼす廃水処理条件の影響2015

    • Author(s)
      片桐誠之、山内一也、入谷英司
    • Organizer
      化学工学会第80年会
    • Place of Presentation
      芝浦工業大学(東京都)
    • Year and Date
      2015-03-19 – 2015-03-21
    • Related Report
      2014 Research-status Report
  • [Presentation] 活性汚泥の沈降特性に及ぼす超音波照射と塩添加の相乗効果2015

    • Author(s)
      片桐誠之、西川匡子、入谷英司
    • Organizer
      化学工学会第80年会
    • Place of Presentation
      芝浦工業大学(東京都)
    • Year and Date
      2015-03-19 – 2015-03-21
    • Related Report
      2014 Research-status Report
  • [Presentation] Evaluation of Cake Properties Based on Variable Pressure Ultrafiltration Associated with Reduction in Cake Surface Area2014

    • Author(s)
      Katagiri, N., R. Nakajima and E. Iritani
    • Organizer
      10th International Conference on Separation Science and Technology (ICSST14)
    • Place of Presentation
      Nara (Japan)
    • Year and Date
      2014-10-30 – 2014-11-01
    • Related Report
      2014 Research-status Report
  • [Presentation] Decolorization Properties of Dye by Enzymatic Membrane Reactor Using Cultural Supernatant of White-Rot fungus2014

    • Author(s)
      Katagiri, N., M. Takatsu and E. Iritani
    • Organizer
      10th International Conference on Separation Science and Technology (ICSST14)
    • Place of Presentation
      Nara (Japan)
    • Year and Date
      2014-10-30 – 2014-11-01
    • Related Report
      2014 Research-status Report
  • [Presentation] Effect of Coexisting Clay on Membrane Filtration Properties of Virus-Containing Suspension2014

    • Author(s)
      Kohori, K., N. Katagiri and E. Iritani
    • Organizer
      10th International Conference on Separation Science and Technology (ICSST14)
    • Place of Presentation
      Nara (Japan)
    • Year and Date
      2014-10-30 – 2014-11-01
    • Related Report
      2014 Research-status Report
  • [Presentation] Cake Properties of Nano-Colloids in Variable Pressure Filtration2014

    • Author(s)
      Iritani, E., N. Katagiri, R. Nakajima and M. Tsukamoto
    • Organizer
      European Conference on Fluid-Particle Separation (FPS) 2014
    • Place of Presentation
      Lyon (France)
    • Year and Date
      2014-10-15 – 2014-10-17
    • Related Report
      2014 Research-status Report
  • [Presentation] Evaluation of Compression-Permeability Characteristics of Microbial Cake Based on Microfiltration Data2014

    • Author(s)
      Katagiri, N., H. Kawahara and E. Iritani
    • Organizer
      European Conference on Fluid-Particle Separation (FPS) 2014
    • Place of Presentation
      Lyon (France)
    • Year and Date
      2014-10-15 – 2014-10-17
    • Related Report
      2014 Research-status Report
  • [Presentation] ウイルスによる粘土質粒子の凝集と膜濾過特性への影響2014

    • Author(s)
      小堀且馬、片桐誠之、入谷英司
    • Organizer
      化学工学会第46回秋季大会
    • Place of Presentation
      九州大学(福岡県)
    • Year and Date
      2014-09-17 – 2014-09-19
    • Related Report
      2014 Research-status Report
  • [Presentation] 白色腐朽菌による酵素産生とその培養液を用いたメンブレンリアクターによる染料脱色2014

    • Author(s)
      高津光伸、片桐誠之、入谷英司
    • Organizer
      化学工学会第46回秋季大会
    • Place of Presentation
      九州大学(福岡県)
    • Year and Date
      2014-09-17 – 2014-09-19
    • Related Report
      2014 Research-status Report
  • [Presentation] エレクトロポレーションシステムを導入したプラスミドDNAのアフィニティ分離法の開発2014

    • Author(s)
      香西真仁、片桐誠之、入谷英司
    • Organizer
      化学工学会第46回秋季大会
    • Place of Presentation
      九州大学(福岡県)
    • Year and Date
      2014-09-17 – 2014-09-19
    • Related Report
      2014 Research-status Report
  • [Presentation] 微生物代謝多糖およびタンパク質の精密濾過特性の評価2014

    • Author(s)
      笹川崇光、片桐誠之、入谷英司
    • Organizer
      化学工学会第46回秋季大会
    • Place of Presentation
      九州大学(福岡県)
    • Year and Date
      2014-09-17 – 2014-09-19
    • Related Report
      2014 Research-status Report
  • [Presentation] Decolorization of Dye Wastewater by Enzymatic Membrane Reactor Using Cultural Supernatant of White-Rot fungus2014

    • Author(s)
      Takatsu, M., N. Katagiri and E. Iritani
    • Organizer
      Particle Separation 2014
    • Place of Presentation
      Sapporo (Japan)
    • Year and Date
      2014-06-15 – 2014-06-18
    • Related Report
      2014 Research-status Report
  • [Presentation] Decolorization Properties of Wastewater by Fungal Laccase-Immobilized Membrane2014

    • Author(s)
      Katagiri, N., Y. Ogi and E. Iritani
    • Organizer
      Singapore International Water Week (SIWW) Water Convention 2014
    • Place of Presentation
      Singapore (Singapore)
    • Year and Date
      2014-06-01 – 2014-06-05
    • Related Report
      2014 Research-status Report
  • [Presentation] 膜ファウリング現象の解析 ケーク濾過・閉塞濾過挙動の解析法2014

    • Author(s)
      片桐誠之
    • Organizer
      先端膜工学研究推進機構平成25年度春季講演会
    • Place of Presentation
      神戸大学(兵庫県)
    • Related Report
      2013 Research-status Report
    • Invited
  • [Presentation] 微生物ケークの圧密特性の評価2014

    • Author(s)
      片桐誠之、河原広隆、入谷英司
    • Organizer
      化学工学会第79年会
    • Place of Presentation
      岐阜大学(岐阜県)
    • Related Report
      2013 Research-status Report
  • [Presentation] 精密濾過で形成される微生物ケークの特性評価2013

    • Author(s)
      片桐誠之、河原広隆、入谷英司
    • Organizer
      分離技術会年会2013
    • Place of Presentation
      日本大学(千葉県)
    • Related Report
      2013 Research-status Report
  • [Presentation] Purification of Plasmid DNA Using Affinity Membrane Filtration2013

    • Author(s)
      Katagiri, N., D. Shimokawa, T. Suzuki and E. Iritani
    • Organizer
      3rd International Conference on Biotechnology Engineering (ICBioE 2013)
    • Place of Presentation
      Kuala Lumpur (Malaysia)
    • Related Report
      2013 Research-status Report
  • [Presentation] 精密濾過試験に基づく微生物ケークの圧縮透過特性の評価2013

    • Author(s)
      河原広隆、片桐誠之、入谷英司
    • Organizer
      化学工学会第45回秋季大会
    • Place of Presentation
      岡山大学(岡山県)
    • Related Report
      2013 Research-status Report
  • [Presentation] 嫌気活性汚泥の膜濾過特性に及ぼす廃水組成の影響2013

    • Author(s)
      山内一也、片桐誠之、入谷英司
    • Organizer
      化学工学会第45回秋季大会
    • Place of Presentation
      岡山大学(岡山県)
    • Related Report
      2013 Research-status Report
  • [Presentation] ウイルス含有汚染懸濁液の膜濾過特性に及ぼす共存粘土の影響2013

    • Author(s)
      小堀且馬、片桐誠之、入谷英司
    • Organizer
      化学工学会第45回秋季大会
    • Place of Presentation
      岡山大学(岡山県)
    • Related Report
      2013 Research-status Report
  • [Presentation] 白色腐朽菌培養液を用いたメンブレンリアクターによる染料の脱色特性2013

    • Author(s)
      高津光伸、片桐誠之、入谷英司
    • Organizer
      化学工学会第45回秋季大会
    • Place of Presentation
      岡山大学(岡山県)
    • Related Report
      2013 Research-status Report
  • [Presentation] Evaluation of Filter Cake Properties in Microfiltration of Microbial Suspensions2013

    • Author(s)
      Katagiri, N., H. Kawahara and E. Iritani
    • Organizer
      Filtech 2013
    • Place of Presentation
      Wiesbaden (Germany)
    • Related Report
      2013 Research-status Report
  • [Presentation] Effect of Cell Morphology on Microfiltration Properties of Microbial Suspensions2013

    • Author(s)
      Katagiri, N., H. Kawahara and E. Iritani
    • Organizer
      Filtration and Separation Symposium 2013
    • Place of Presentation
      Tokyo (Japan)
    • Related Report
      2013 Research-status Report
  • [Book] 吸着・分離材料の設計、性能評価と新しい応用(膜閉塞・ケーク濾過挙動の解析(第5章 シミュレーションによる吸着,分離挙動の把握,性能予測, 6節))2015

    • Author(s)
      片桐誠之
    • Total Pages
      524
    • Publisher
      技術情報協会
    • Related Report
      2015 Annual Research Report
  • [Book] 粉体工学ハンドブック(ろ過・圧搾(第3章 粉体を扱う単位操作, 3.9))2014

    • Author(s)
      入谷英司
    • Total Pages
      776
    • Publisher
      朝倉書店
    • Related Report
      2013 Research-status Report
  • [Remarks] 分離プロセス工学研究グループ

    • URL

      http://www.nuce.nagoya-u.ac.jp/L5/

    • Related Report
      2015 Annual Research Report
  • [Remarks] 分離プロセス工学研究グループ

    • Related Report
      2014 Research-status Report
  • [Remarks] http://www.nuce.nagoya-u.ac.jp/L5/

    • Related Report
      2014 Research-status Report
  • [Remarks] 分離プロセス工学研究グループ

    • URL

      http://www.nuce.nagoya-u.ac.jp/L5/

    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2022-02-01  

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