Construction of human antibody library and establishment of selection method to develop fragment antibody therapeutics

• Project/Area Number: 21390032
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Drug development chemistry
• Research Institution: Kagoshima University
• Principal Investigator: ITO Yuji 鹿児島大学, 大学院・理工学研究科, 教授 (60223195)
• Co-Investigator(Kenkyū-buntansha): ARIMA Naomichi 鹿児島大学, 大学院・医歯学総合研究科, 教授 (30175997) ; KUROKI Ryota 独立行政法人日本原子力研究開発機構, 量子ビーム応用研究部門, ユニット長 (30391246)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥14,820,000 (Direct Cost: ¥11,400,000、Indirect Cost: ¥3,420,000); Fiscal Year 2011: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2010: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000); Fiscal Year 2009: ¥7,670,000 (Direct Cost: ¥5,900,000、Indirect Cost: ¥1,770,000)
• Keywords: 医薬分子機能学 / 抗体 / 低分子化抗体 / T7ファージ / ファージ提示法 / 安定性

Research Abstract

In this study, we constructed the libraries of human single-chain Fv antibody and llama VHH antibody using T7 phage display system and established the selection method to isolate the specific phages. Especially, we succeeded in the isolation of the T7 phage clones binding to the antigens from the library, which indicates our library system is useful for a rapid generation of specific antibodies. On the other hand, we found the problematic issues that the non-specific binding phages were enriched by selection process because of folding problems and instability of antibody, which should be solved in the further investigations.

Research Products

• [Journal Article] Isolation and Affinity Maturation of Human IgA-Specific Peptide from Random Peptide T7 Phage Library for Designing a New IgA Affinity Ligand (2012)
  • Author(s): Hatanaka T., Tsukamoto S., Sakamoto K., Arima K., Ito Y.
  • Journal Title: Peptide Science 2011 Pages: 25-26
  • NAID: 10030203951
  • Peer Reviewed
• [Journal Article] A mimotope peptide of Aβ42 fibril-specific antibodies with Aβ42 fibrillation inhibitory activity induces anti-Abeta42 conformer antibody response by a displayed form on an M13 phage in mice (2011)
  • Author(s): Tanaka, K., Nishimura, M., Yamaguchi, Y., Hashiguchi, S., Takiguchi, S., Yamaguchi, M., Tahara, H., Gotanda, T., Abe, R., Ito, Y., Sugimura, K.
  • Journal Title: Journal of neuroimmunology Volume: 236 Pages: 27-38
  • DOI: 10.1016/j.jneuroim.2011.04.010
  • Peer Reviewed
• [Journal Article] A mimotope peptide of Abeta42 fibril-specific antibodies with Abeta42 fibrillation inhibitory activity induces anti-Abeta42 conformer antibody response by a displayed form on an M13 phage in mice (2011)
  • Author(s): Tanaka, K., Nishimura, M., Yamaguchi, Y., Hashiguchi, S., Takiguchi, S., Yamaguchi, M., Tahara, H., Gotanda, T., Abe, R., Ito, Y., Sugimura, K.
  • Journal Title: J Neuroimmunol Volume: 236 Pages: 27-38
  • Peer Reviewed
• [Journal Article] Characterization of human single-chain antibodies against highly pathogenic avian influenza H5N1 viruses : Mimotope and Neutralizing Activity (2010)
  • Author(s): Yang, J., Yoshida, R., Kariya, Y., Zhang, X., Hashiguchi, S., Nakashima, T., Suda, Y., Takada, A., Ito, Y., Sugimura, K.
  • Journal Title: J Biochem Volume: 148 Pages: 507-515
  • DOI: 10.1093/jb/mvq084
  • NAID: 10027879101
  • Peer Reviewed
• [Journal Article] ファージディスプレイBeyond antibody:～抗体様分子による分子標的～ (2010)
  • Author(s): 橋口周平、伊東祐二、田中孝一、松木園美穂、村岡賢、杉村和久
  • Journal Title: 生化学 Volume: 82 Pages: 710-726
• [Journal Article] Short communication : generation of recombinant monoclonal antibodies against an immunodominant HLA-A 2402-restricted HIV type 1 CTL epitope (2009)
  • Author(s): Nunoya, J., Nakashima, T., Kawana-Tachikawa, A., Kiyotani, K., Ito, Y., Sugimura, K., Iwamoto, A.
  • Journal Title: AIDS research and human retroviruses Volume: 25 Pages: 897-904
  • Peer Reviewed
• [Journal Article] Effective induction of cell death on adult T-cell leukaemia cells by HLA-DRbeta-specific small antibody fragment isolated from human antibody phage library (2009)
  • Author(s): Muraoka, S., Ito, Y., Kamimura, M., Baba, M., Arima, N., Suda, Y., Hashiguchi, S., Torikai, M., Nakashima, T., Sugimura, K.
  • Journal Title: Journal of biochemistry Volume: 145 Pages: 799-810
  • DOI: 10.1093/jb/mvp039
  • Peer Reviewed
• [Journal Article] Regulation of T cell response by blocking the ICOS signal with the B7RP-1-specific small antibody fragment isolated from human antibody phage library (2009)
  • Author(s): Maeda, M., Ito, Y., Hatanaka, T., Hashiguchi, S., Torikai, M., Nakashima, T., Sugimura, K.
  • Journal Title: MAbs Volume: 1 Pages: 453-461
  • URL: http://www.landesbioscience.com/journals/mabs/9633
  • Peer Reviewed
• [Journal Article] Regulation of T cell response by blocking the ICOS signal with the B7RP-1-specific small antibody fragment isolated from human antibody phage library (2009)
  • Author(s): Maeda, M., Ito, Y., Hatanaka, T., Hashiguchi, S., Torikai, M., et al.
  • Journal Title: mAb 1 Pages: 450-457
  • Peer Reviewed
• [Journal Article] Discovery and characterization of a peptide motif that specifically recognizes a non-native conformation of human IgG induced by acidic pH conditions (2009)
  • Author(s): Sakamoto K., Ito Y., Hatanaka T., et al.
  • Journal Title: The Journal of biological chemistry 284 Pages: 9986-9993
  • Peer Reviewed
• [Journal Article] Effective Induction of Cell Death on Adult T-Cell Leukemia Cells by HLA-DR{beta}-Specific Small Antibody Fragment Isolated From Human Antibody Phage Library (2009)
  • Author(s): Muraoka S., Ito Y., Kamimura M., Baba M., Arima N., Suda Y., et al.
  • Journal Title: Journal of biochemistry 145 Pages: 799-810
  • Peer Reviewed
• [Presentation] ファージディスプレイ技術による機能性ペプチド・抗体のデザインと応用 (2012)
  • Author(s): 伊東祐二
  • Organizer: 第11回化学・材料研究セミナー
  • Place of Presentation: 福岡
  • Year and Date: 2012-01-07
• [Presentation] Peptide designs for affinity ligands of immunoglobulin by random peptide phage library (2011)
  • Author(s): Yuji Ito
  • Organizer: 15^<th> Korean Peptide and Protein Symposium
  • Place of Presentation: Ochag (Korea)(招待講演)
  • Year and Date: 2011-12-01
• [Presentation] T7ファージディスプレイによる抗体ライブラリの構築と評価 (2011)
  • Author(s): 塚本翔悟, 畠中孝彰, 有馬一成, 伊東祐二
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2011-09-21
• [Presentation] T7ファージディスプレイによる抗体ライブラリの構築と評価 (2011)
  • Author(s): 塚本翔悟、畠中孝彰、有馬一成、伊東祐二
  • Organizer: 第84回日本生化学会
  • Place of Presentation: 京都市
  • Year and Date: 2011-09-21
• [Presentation] T7ファージライブラリを用いたヒト抗体結合ペプチドのデザインと評価,第4回生体分子科学シンポジウム (2011)
  • Author(s): 伊東祐二
  • Organizer: 第2回ケミカルバイオロジー研究所シンポジウム
  • Place of Presentation: 大阪
  • Year and Date: 2011-03-04
• [Presentation] ファージライブラリによるヒト抗体結合性ペプチドリガンドの探索と応用 (2011)
  • Author(s): 伊東祐二
  • Organizer: 第4回 生体分子科学シンポジウム
  • Place of Presentation: 大阪
  • Year and Date: 2011-03-04
• [Presentation] Tファージ提示法による機能性抗体とペプチドのデザイン (2010)
  • Author(s): 伊東祐二
  • Organizer: 大阪大学蛋白質研究所セミナー「バクテリオファージ研究の可能性と課題
  • Place of Presentation: 大阪
  • Year and Date: 2010-09-10
• [Presentation] 高機能性低分子化抗体の単離を目指したT7ファージによる抗体ライブラリの構築 (2010)
  • Author(s): 塚本翔悟, 畠中孝彰, 有馬一成, 伊東祐二
  • Organizer: 平成22年度日本生化学会九州支部例会
  • Place of Presentation: 鹿児島
  • Year and Date: 2010-05-22
• [Presentation] Selections of cell-specific antibodies and Ig-specific peptides by phage display : their potentials for clinical and industrial uses (2010)
  • Author(s): Ito Y
  • Organizer: 2010 KSBB (Korean Society for Biotechnology and Bopscience) Spring Meeting and International Symposium
  • Place of Presentation: ソウル(韓国)
  • Year and Date: 2010-04-16
• [Presentation] T7ファージライブラリを用いたヒト抗体結合ペプチドのデザインと評価 (2009)
  • Author(s): 伊東祐二
  • Organizer: 第82回日本生化学会大会シンポジウム:Beyond antibody という研究領域
  • Place of Presentation: 神戸ポートアイランド(神戸市)
  • Year and Date: 2009-10-21
• [Presentation] ペプチドライブラリを用いたヒト抗体結合ペプチドのデザインと評価 (2009)
  • Author(s): 伊東祐二
  • Organizer: 第9回日本蛋白質科学会年会
  • Place of Presentation: 熊本全日空ホテルニュースカイ(熊本市)
  • Year and Date: 2009-05-20
• [Book] ファージディスプレイBeyond antibody :～抗体様分子による分子標的～ (2010)
  • Author(s): 橋口周平、伊東祐二、田中孝一、松木園美穂、村岡賢、杉村和久
  • Publisher: 生化学
• [Patent(Industrial Property Rights)] IgG結合性ペプチド (2012)
  • Inventor(s): 伊東祐二
  • Industrial Property Rights Holder: 鹿児島大学
  • Acquisition Date: 2012-03-23
  • Acquisition Year: 2012
• [Patent(Industrial Property Rights)] IgA結合性ペプチド及びそれによるIgAの精製 (2011)
  • Inventor(s): 伊東祐二、伊東治、大薗慎二、石飛宏幸
  • Industrial Property Rights Holder: 鹿児島大学、大塚化学(株)
  • Industrial Property Number: 2011-262871
  • Filing Date: 2011-11-30
  • Application Year: 2011
• [Patent(Industrial Property Rights)] IgG結合性ペプチド及びそれによるIgGの検出および精製方法 (2011)
  • Inventor(s): 伊東祐二
  • Industrial Property Rights Holder: 鹿児島大学、大塚化学株式会社
  • Industrial Property Number: 2011-182539
  • Filing Date: 2011-08-24
  • Application Year: 2011
• [Patent(Industrial Property Rights)] IgG結合性ペプチド及びそれによるIgGの検出および精製方法 (2010)
  • Inventor(s): 伊東祐二
  • Industrial Property Rights Holder: 鹿児島大学、大塚化学(株)
  • Industrial Property Number: 2011-182539
  • Filing Date: 2010-08-24
  • Application Year: 2010
• [Patent(Industrial Property Rights)] 特許出願 (2010)
  • Inventor(s): 伊東祐二
  • Industrial Property Rights Holder: 鹿児島大学・大塚化学
  • Filing Date: 2010-05-24
  • Application Year: 2010