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Development of anti-cancer antibodies through high-throughput sequencing from antibody phage library

Research Project

Project/Area Number 25460151
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Drug development chemistry
Research InstitutionKagoshima University

Principal Investigator

ITO YUJI  鹿児島大学, 理工学研究域理学系, 教授 (60223195)

Co-Investigator(Kenkyū-buntansha) ARIMA Naomichi  鹿児島大学, 医歯学総合研究科, 客員研究員 (30175997)
Project Period (FY) 2013-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Keywords抗体医薬 / ガン / ファージライブラリー / 次世代シークエンサー / バイオパンイング / 単鎖Fv抗体 / ヒト抗体 / 抗体 / 治療 / 細胞 / ファージライブラリ / バイオパンニング / ガン細胞 / 網羅的配列解析 / 抗体ライブラリ
Outline of Final Research Achievements

In this study, we aimed to establish the short-term and reliable system for isolating anti-cancer antibodies (single chain Fv antibodies) from antibody phage library, especially by combining the biopanning and the comprehensive sequence analysis on next generation sequencer (NGS method). As a result, we successfully isolated several clones of lutheran and antigen A-specific antibodies which targeted hepatic and Esophageal cancers, respectively. These results indicate that NGS method is more useful in isolation of multiple antigen-specific antibodies, as compared with the conventional biopanning method comprising of biopanning and cloning/binding screening processes.

Report

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

    (28 results)

All 2016 2015 2014 2013 Other

All Journal Article (10 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 8 results,  Open Access: 4 results,  Acknowledgement Compliant: 2 results) Presentation (12 results) (of which Int'l Joint Research: 1 results,  Invited: 5 results) Book (1 results) Remarks (2 results) Patent(Industrial Property Rights) (3 results)

  • [Journal Article] Selection of Color-Changing and Intensity-Increasing Fluorogenic Probe as Protein-Specific Indicator Obtained via the 10BASEd-T2016

    • Author(s)
      Masumi Taki, Hiroaki Inoue, Kazuto Mochizuki, Jay Yang, and Yuji Ito
    • Journal Title

      Anal. Chem.

      Volume: 88 Issue: 2 Pages: 1096-1099

    • DOI

      10.1021/acs.analchem.5b04687

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] 特集抗体医薬の進歩“ヒトIgG特異的化学修飾による機能性抗体医薬の創出”2016

    • Author(s)
      伊東祐二
    • Journal Title

      細胞

      Volume: 48 Pages: 172-176

    • Related Report
      2015 Annual Research Report
  • [Journal Article] Isolation and characterization of antigen-specific alpaca (Lama pacos) VHH antibodies by biopanning followed by high throughput sequencing2015

    • Author(s)
      Nobuo Miyazaki, Norihiko Kiyose, Yoko Akazawa, Mizuki Takashima, Yosihisa Hagihara, Naokazu Inoue, Tomonari Matsuda, Ryu Ogawa, Seiya Inoue, Yuji Ito
    • Journal Title

      Journal of Biochemistry

      Volume: 157 Issue: 3 Pages: 205-215

    • DOI

      10.1093/jb/mvv038

    • Related Report
      2015 Annual Research Report 2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] 分子標的素子デザインにおけるファージライブラリー法2015

    • Author(s)
      橋口周平, 宮原隆二, 岸本聡, 伊東祐二
    • Journal Title

      生物工学

      Volume: 93 Pages: 289-292

    • Related Report
      2015 Annual Research Report
    • Open Access
  • [Journal Article] Highly efficient production of VHH antibody fragments in Brevibacillus choshinensis expression system2015

    • Author(s)
      Makoto Mizukami, Hiroko Tokunaga, Hiromasa Onishi, Yohei Ueno, Hiroshi Hanagata, Nobuo Miyazaki, Norihiko Kiyose, Yuji Ito, Matsujiro Ishibashi, Yoshihisa Hagihara, Tsutomu Arakawa, Akira Miyauchi, Masao Tokunaga
    • Journal Title

      Protein Expression and Purification

      Volume: 105 Pages: 23-32

    • DOI

      10.1016/j.pep.2014.09.017

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Selection and Characterization of Human Serum Albumin-specific Porcine scFv Antibodies, Using a Phage Display Library2014

    • Author(s)
      Junko Muraoka, Takuya Ozawa, Yurie Enomoto, Norihiko Kiyose, Ayana Imamura, Kazunari Arima, Hiroshi Nakayama, and Yuji Ito
    • Journal Title

      Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

      Volume: 33 Issue: 1 Pages: 42-66

    • DOI

      10.1089/mab.2013.0068

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Construction of a crown ether-like supramolecular library by conjugation of genetically-encoded peptide linkers displayed on bacteriophage T72014

    • Author(s)
      Keisuke Fukunaga, Takaaki Hatanaka, Yuji Ito, Michiko Minami and Masumi Taki
    • Journal Title

      Chem. Commun.

      Volume: 50 Issue: 30 Pages: 3921-3923

    • DOI

      10.1039/c4cc00811a

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Pharmacophore Generation from a Drug-like Core Molecule Surrounded by a Library Peptide via the 10BASEd-T on Bacteriophage T72014

    • Author(s)
      Yuuki Tokunaga, Yuuki Azetsu, Keisuke Fukunaga, Takaaki Hatanaka, Yuji Ito and Masumi Taki
    • Journal Title

      Molecules

      Volume: 19 Issue: 2 Pages: 2481-2496

    • DOI

      10.3390/molecules19022481

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Gp10 based-thioetherification (10BASEd-T) on a displaying library peptide of bacteriophage T72013

    • Author(s)
      Keisuke Fukunaga, Takaaki Hatanaka, Yuji Ito and Masumi Taki
    • Journal Title

      Mol. Biosyst.

      Volume: 9 Issue: 12 Pages: 2988-2991

    • DOI

      10.1039/c3mb70379g

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Journal Article] Preparation and evaluation of cellulose-dissolving magnetic ionic liquid2013

    • Author(s)
      Muraoka, J., Kamiya, N., and Ito, Y
    • Journal Title

      Journal of Molecular Liquids

      Volume: 182 Pages: 76-78

    • DOI

      10.1016/j.molliq.2013.03.012

    • NAID

      120005303831

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Presentation] 網羅的配列解析による多様な候補抗体の迅速同定手法の開発2016

    • Author(s)
      伊東祐二
    • Organizer
      日本薬学会第136年会
    • Place of Presentation
      横浜コンベンションセンター(神奈川・横浜)
    • Year and Date
      2016-03-27
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 革新的バイオ医薬品創製に向けたファージライブラリによる機能性抗体・ペプチドのデザイン2015

    • Author(s)
      伊東祐二
    • Organizer
      日本薬学会九州支部主催 特別講演会
    • Place of Presentation
      長崎大学薬学部(長崎・長崎)
    • Year and Date
      2015-12-14
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 次世代シークエンサーを用いた網羅的配列解析によるファージライブラリからの特異的抗体の効率的同定法2015

    • Author(s)
      伊東祐二
    • Organizer
      生物化学的測定研究会第20回学術シンポジウム
    • Place of Presentation
      神戸薬科大学(兵庫・神戸)
    • Year and Date
      2015-11-13
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 次世代シークエンサー配列解析による効率的な抗原特異的抗体の同定2015

    • Author(s)
      伊東祐二
    • Organizer
      第67回日本生物工学会シンポジウム
    • Place of Presentation
      城山ホテル(鹿児島・鹿児島)
    • Year and Date
      2015-10-26
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] 革新的バイオ医薬創製に向けたファージライブラリによる機能性抗体・ペプチドのデザイン2015

    • Author(s)
      伊東祐二
    • Organizer
      第21 回ペプチドフォーラム
    • Place of Presentation
      東京薬科大学千代田サテライトキャンパス(東京・千代田区)
    • Year and Date
      2015-08-29
    • Related Report
      2015 Annual Research Report
    • Invited
  • [Presentation] Identification of disease related antigen-specific novel human antibodies by method that combines biopanning and analysis of next generation sequencer from patient-derived scFv antibody library2015

    • Author(s)
      Yuji Ito, Yurie Enomoto, Shuhei Umemura, Aiko Fujiyama, Ryoko Mieno, Yukiko Kato and Dai-ichiro Kato
    • Organizer
      The 29th annual symposium of the protein society
    • Place of Presentation
      Barcelona(Spain)
    • Year and Date
      2015-07-22
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 次世代シークエンサーを用いた網羅的抗体配列解析によるルテラン特異的な肝癌患者由来抗体の解析2014

    • Author(s)
      藤山愛子、榎元友里恵、吉川大和、Hui Kam Man、松田知成、伊東祐二
    • Organizer
      平成26年度日本生化学会九州支部例会
    • Place of Presentation
      九州大学病院キャンパス コラボステーション I(福岡県福岡市)
    • Year and Date
      2014-05-17 – 2014-05-18
    • Related Report
      2014 Research-status Report
  • [Presentation] Isolation and characterization of antigen-specific VHH antibodies from immunized alpaca VHH antibody phage lbrary using Next Generation Sequencing (NGS)2013

    • Author(s)
      Nobuo Miyazaki, Norihiko Kiyose, Yosihisa Hagihara, Matsuda Tomonari, Seiya Inoue, Kzunori Arima and Yuji Ito
    • Organizer
      Antibody Engineering & Therapeutics 2013
    • Place of Presentation
      Huntington Beach, CA
    • Related Report
      2013 Research-status Report
  • [Presentation] 患者由来の抗体ファージライブラリから得られた肝癌細胞抗原ルテランに対する自己抗体の性状解析2013

    • Author(s)
      3) 榎元友里恵,吉川大和,Hui Kam Man,有馬一成,伊東祐二
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report
  • [Presentation] アルパカ由来VHHナイーブ抗体ファージライブラリから単離された抗原特異的VHH抗体の単離と機能解析2013

    • Author(s)
      4) 清瀬紀彦,宮崎誠生,井上聖也,有馬一成,岸本聡,萩原義久,伊東祐二
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report
  • [Presentation] 抗原免疫動物から構築したアルパカVHH抗体ファージライブラリからの抗原特異的抗体の単離と機能解析2013

    • Author(s)
      6) 宮﨑誠生,清瀬紀彦,萩原義久,井上聖也,有馬一成,伊東祐二
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report
  • [Presentation] ブタ抗体ファージライブラリを使った抗原特異的ヒト化抗体2013

    • Author(s)
      小澤拓矢,若井純子,有馬一成,伊東祐二
    • Organizer
      第86回日本生化学会大会
    • Place of Presentation
      横浜
    • Related Report
      2013 Research-status Report
  • [Book] 免疫測定法 基礎から先端まで(小林典弘、他、編集)2014

    • Author(s)
      伊東祐二、宮崎誠生、榎元友里恵
    • Total Pages
      10
    • Publisher
      講談社
    • Related Report
      2014 Research-status Report
  • [Remarks] 研究者総覧(伊東祐二)

    • URL

      http://kuris.cc.kagoshima-u.ac.jp/705524.html

    • Related Report
      2014 Research-status Report
  • [Remarks] 研究室ホームページ(伊東研究室)

    • URL

      http://www.sci.kagoshima-u.ac.jp/~yito/

    • Related Report
      2014 Research-status Report
  • [Patent(Industrial Property Rights)] 抗癌剤2015

    • Inventor(s)
      吉川大和, 野水基義, 福原武志 , 伊東祐二
    • Industrial Property Rights Holder
      吉川大和, 野水基義, 福原武志 , 伊東祐二
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2015-230926
    • Filing Date
      2015-11-26
    • Related Report
      2015 Annual Research Report
  • [Patent(Industrial Property Rights)] IgG結合ペプチドによる抗体の特異的修飾2015

    • Inventor(s)
      伊東祐二
    • Industrial Property Rights Holder
      伊東祐二
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2015-103153
    • Filing Date
      2015-05-20
    • Related Report
      2015 Annual Research Report
  • [Patent(Industrial Property Rights)] 低分子化抗体のスクリーニング方法及び製造方法2013

    • Inventor(s)
      伊東 祐二、他5名
    • Industrial Property Rights Holder
      伊東 祐二、他5名
    • Industrial Property Rights Type
      特許
    • Industrial Property Number
      2013-241806
    • Filing Date
      2013-11-29
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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