Development of rapid generation of antibodies with high affinity for improvement of detection limi

• Project/Area Number: 23790617
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Laboratory medicine
• Research Institution: University of Toyama
• Principal Investigator: OZAWA Tatsuhiko 富山大学, 大学院・医学薬学研究部(医学), 助教 (10432105)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2011: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: 臨床検査医学 / 高親和性抗体 / ウサギモノクローナル抗体 / ISAAC / 抗体医薬

Research Abstract

Antigen-specific rabbit monoclonal antibodies （RaMoAbs) are useful due to their high specificity and high affinity, and the establishment of a comprehensive and rapid RaMoAb generation system has been highly anticipated. Here, we present a novel system for rapid RaMoAb generation and produce antigen-specific RaMoAbs with high affinity within a time period of only 7 days. We have used this system to efficiently generate RaMoAbs that are specific to a phosphorylated signal-transducing molecule. Our system provides a new method for the comprehensive and rapid production of RaMoAbs, which may contribute to laboratory research and clinical applications.交付決定額

Research Products

• [Journal Article] Reduced Level of the BCL11B Protein Is Associated with Adult T-Cell Leukemia/Lymphoma (2013)
  • Author(s): Kurosawa, N.
  • Journal Title: PLoS One Volume: 8 Issue: 1 Pages: e55147-e55147
  • DOI: 10.1371/journal.pone.0055147
  • Peer Reviewed
• [Journal Article] A novel rabbit immunospot array assay on a chip allows for the rapid generation of rabbit monoclonal antibodies with high affinity. (2012)
  • Author(s): Ozawa T., et al
  • Journal Title: PLoS One Volume: 7 Issue: 12 Pages: e52383-e52383
  • DOI: 10.1371/journal.pone.0052383
  • Peer Reviewed
• [Journal Article] The binding affinity of a soluble TCR-Fc fusion protein is significantly improved by crosslinkage with an anti-Cbeta antibody (2012)
  • Author(s): Ozawa T, Horii M, Kobayashi E, Jin A, Kishi H, Muraguchi A
  • Journal Title: Biochem Biophys Res Commun Volume: 422 Pages: 245-249
  • Peer Reviewed
• [Journal Article] Unbiased Analysis of TCRα/β Chains at the Single-Cell Level in Human CD8^<+> T-Cell Subsets (2012)
  • Author(s): Sun X, Saito M, Sato Y, Chikata T, Naruto T, Ozawa T, Kobayashi E, Kishi H, Muraguchi A, Takiguchi M
  • Journal Title: PLoS One Volume: 7 Issue: 7 Pages: e40386-e40386
  • DOI: 10.1371/journal.pone.0040386
  • Peer Reviewed
• [Journal Article] HELIOS-BCL11B fusion gene involvement in a t(2;14)(q34;q32) in an adult T-cell leukemia patient (2012)
  • Author(s): Fujimoto R, Ozawa T, Itoyama T, Sadamori N, Kurosawa N, Isobe M
  • Journal Title: Cancer Genet Volume: 205 Pages: 356-364
  • Peer Reviewed
• [Journal Article] The binding affinity of a soluble TCR-Fc fusion protein is significantly improved by crosslinkage with an anti-Cβ antibody (2012)
  • Author(s): Ozawa T ., Horii M., Kobayashi E., Jin A., Kishi H., and Muraguchi A
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 422 Issue: 2 Pages: 245-249
  • DOI: 10.1016/j.bbrc.2012.04.134
  • Peer Reviewed
• [Journal Article] Characterization of a fully human monoclonal antibody against extracellular domain of matrix protein 2 of influenza A virus. (2011)
  • Author(s): Ozawa T, Jin A, Tajiri K, Takemoto M, Okuda T, Shiraki K, Kishi H, Muraguchi A
  • Journal Title: Antiviral Res Volume: 91 Pages: 283-287
  • Peer Reviewed
• [Journal Article] Characterization of a fully human monoclonal antibody against extracellular domain of matrix protein 2 of influenza A virus (2011)
  • Author(s): Ozawa T, et al
  • Journal Title: Antiviral Res Volume: 91 Pages: 283-287
  • Peer Reviewed
• [Journal Article] Rapid isolation of antigen-specific antibody-secreting cells using a chip-based immunospot array (2011)
  • Author(s): JIn A, et al
  • Journal Title: Nat Protoc Volume: 6 Pages: 668-676
  • Peer Reviewed
• [Presentation] Rabbit ISAAC (rabbit immunospot array assay on a chip) allows for the rapid generation of rabbit monoclonal antibodies with high affinity (2013)
  • Author(s): Piao X, Muraguchi A, Hamana H, Ozawa T, Kobayashi E, Jin A, Kishi H
  • Organizer: Keystone Symposia (Antibodies as Drugs)
  • Place of Presentation: Vancouver, British Columbia, Canada
• [Presentation] Rabbit ISAAC (rabbit immunospot array assay on a chip) allows for the rapid generation of rabbit monoclonal antibodies with high affinity (2013)
  • Author(s): Hamana H
  • Organizer: Keystone Symposia (Antibodies as Drugs)
  • Place of Presentation: Vancouver, Canada
• [Presentation] A novel rabbit immunospot array assay on a chip allows for the rapid generation of rabbit monoclonal antibodies (2012)
  • Author(s): Ozawa T, Piao X, Kobayashi E, Zhou Y, Sakurai H, Andoh T, Jin A, Kishi H.Muraguchi A
  • Organizer: 第34回日本分子生物学会年会
  • Place of Presentation: 福岡
• [Presentation] The binding affinity of a soluble TCR-Fc fusion protein is significantly improved by crosslinkage with an anti-Cβ antibody. (2012)
  • Author(s): Ozawa T, Kobayashi E, Jin A, Kishi H, Muraguchi A
  • Organizer: 第41回日本免疫学会総会・学術集会
  • Place of Presentation: 神戸
• [Presentation] Rapid generation of super-antibodies using rabbit ISAAC (2012)
  • Author(s): Piao X, Ozawa T, Kobayashi E, Kishi H, Muraguchi A.
  • Organizer: 第41回日本免疫学会総会・学術集会
  • Place of Presentation: 神戸
• [Presentation] Cloning of human antigen-specific TCRs can confer the candidates for cancer gene therapy (2012)
  • Author(s): Kobayashi E, Kishi H, Hamana H, Ozawa T, Nakagawa H, Jin A, Muraguchi A.
  • Organizer: 第41回日本免疫学会総会・学術集会
  • Place of Presentation: 神戸
• [Presentation] Detection of antigen-stimulated cytokine-secretion in human T-cells at single cell levels on a live cell chip. (2012)
  • Author(s): Kishi H, Jin A, Kobayashi E, Ozawa T, Hamana H, Muraguchi A.
  • Organizer: Detection of antigen-stimulated cytokine-secretion in human T-cells at single cell levels on a live cell chip.
  • Place of Presentation: 神戸
• [Presentation] Comparison of TCR repertoires in EBV-specific T cells detected by three different staining methods. (2012)
  • Author(s): Hamana H, Kobayashi E, Kishi H, Ozawa T, Jin A, Muraguchi A
  • Organizer: 第41回日本免疫学会総会・学術集会
  • Place of Presentation: 神戸
• [Presentation] Analysis of cellular responses at singel cell levels with a hybrid magnetic microwell array chip (2012)
  • Author(s): Kishi H, Obata T, Takami S, Kazui M, Ozawa T, Ogawa A, Muraguchi A
  • Organizer: International Joint Symposium on Single-Cell Analysis
  • Place of Presentation: 京都
• [Presentation] Cloning of human antigen-specific TCRs can confer the candidates for cancer gene therapy (2012)
  • Author(s): Kobayashi E, Mizukoshi E, Kishi H, Hamana H, Nagai T, Ozawa T, Nakagawa H, Jin A, Kaneko S, Muraguchi A
  • Organizer: SITC 27th Annual Meeting
  • Place of Presentation: North Bethesda, MD, USA
• [Presentation] がんのTCR遺伝子治療にむけた候補TCR遺伝子の迅速クローニング法 (2012)
  • Author(s): 岸裕幸, 小林栄治, 水腰英四郎, 小澤龍彦, 金子周一, 村口篤
  • Organizer: 第71回日本癌学会学術集会
  • Place of Presentation: 札幌
• [Presentation] he binding affinity of a soluble TCR-Fc fusion protein is significantly improved by crosslinkage with an anti-Cβ antibody (2012)
  • Author(s): Ozawa T
  • Organizer: 第41回日本免疫学会総会・学術集会
  • Place of Presentation: 神戸
• [Presentation] Detection of antigen-stimulated cytokine-secretion in human T-cells at single cell levels on a live cell chip (2012)
  • Author(s): Kishi H
  • Organizer: 第41回日本免疫学会総会・学術集会
  • Place of Presentation: 神戸
• [Presentation] Comparison of TCR repertoires in EBV-specific T cells detected by three different staining methods (2012)
  • Author(s): Hamana H
  • Organizer: 第41回日本免疫学会総会・学術集会
  • Place of Presentation: 神戸
• [Presentation] Cloning of human antigen-specific TCRs can confer the candidates for cancer gene therapy (2012)
  • Author(s): Kobayashi E
  • Organizer: SITC 27th Annual Meeting
  • Place of Presentation: North Bethesda, USA
• [Presentation] A high-throughput method for generating rabbit monoclonal antibodies by immunospot array assay on a chip technology (2011)
  • Author(s): Ozawa T, Piao X, Kobayashi E, Kishi H, Muraguchi A.
  • Organizer: 第33回日本分子生物学会年会
  • Place of Presentation: 横浜
• [Presentation] 可溶化TCRの作製と機能解析 (2011)
  • Author(s): 小澤龍彦, 小林栄治, 岸裕幸, 村口篤
  • Organizer: 第40回日本免疫学会総会・学術集会
  • Place of Presentation: 千葉
• [Presentation] The novel method to detect single antibody-secreting B-cells and single cytokine-secreting T-cells using a cell-microarray chip (2011)
  • Author(s): Ozawa T, Kobayashi E, Tajiri K, Jin A, Kishi H, Muraguchi A
  • Organizer: Cold Spring Harbor Meeting, Single cell analysis
  • Place of Presentation: New York, USA
• [Presentation] 単一細胞解析に基づく迅速な抗原特異的T細胞受容体クローニングシステム. (2011)
  • Author(s): Kobayashi E, Hamana H, Nagai T, Ozawa T, Kishi H, Muraguchi A.
  • Organizer: 第40回日本免疫学会総会・学術集会
  • Place of Presentation: 千葉
• [Presentation] 迅速なTCR cDNAクローニングシステムを用いたEBV特異的単一T細胞のTCRα/βレパトア解析 (2011)
  • Author(s): 浜名洋, 小林栄治, 長井輝美, 小澤龍彦, 岸裕幸, 村口篤
  • Organizer: 第40回日本免疫学会総会・学術集会
  • Place of Presentation: 千葉
• [Presentation] ISAAC法を用いた迅速かつハイスループットなウサギモノクローナル抗体作成法の開発 (2011)
  • Author(s): 朴秀虹, 小澤龍彦, 小林栄治, 岸裕幸, 村口篤.
  • Organizer: 第40回日本免疫学会総会・学術集会
  • Place of Presentation: 千葉
• [Presentation] 細胞チップを用いた抗原特異的サイトカイン分泌T細胞の単一細胞レベルでの検出 (2011)
  • Author(s): Kishi H, Kobayashi E, Ozawa T, Hamana H, Nagai T, Tajiri K, Muraguchi A
  • Organizer: 第40回日本免疫学会総会・学術集会
  • Place of Presentation: 千葉
• [Presentation] Single T cell analysis system for rapid cloning and functional evaluation of antigen-specific T cell receptors Cold Spring Harbor Conferences Asia (2011)
  • Author(s): Kobayashi E, Hamana H, Nagai T, Horii M, Ozawa T, Kishi H, Muraguchi A.
  • Place of Presentation: Suzhou, China.
• [Presentation] 単一細胞解析に基づく迅速な抗原特異的T細胞受容体クローニングシステム (2011)
  • Author(s): Kobayashi E, et al
  • Organizer: 第40回日本免疫学会総会・学術集会
  • Place of Presentation: 千葉
• [Presentation] Single T cell analysis system for rapid cloning and functional evaluation of antigen-specific T cell receptors (2011)
  • Author(s): Kobayashi E, et al
  • Organizer: Cold Spring Harbor Conferences Asia
  • Place of Presentation: Suzhou, China
• [Book] Screening of antigen-specific antibody-secreting cells (2012)
  • Author(s): Kishi H, Jin A, Ozawa T, Tajiri K, Obata T, Muraguchi A
  • Publisher: Springer、Methods Mol Biol
• [Book] Screening of antigen-specific antibody-secreting cells (2012)
  • Author(s): Kishi H. et al
  • Total Pages: 10
  • Publisher: Humana Press「Methods Mol Biol」
• [Book] ンパ球アレイ法を用いた抗体医薬の開発. 細胞 (2011)
  • Author(s): 小澤龍彦, 田尻和人, 岸裕幸, 村口篤
  • Publisher: ニューサイエンス社
• [Book] シングル細胞チップと抗体医薬への応用 (2011)
  • Author(s): 小澤龍彦, 岸裕幸, 村口篤
  • Publisher: シーエムシー出版、ナノ融合による先進バイオデバイス
• [Book] 新しい抗体スクリーニング法. (2011)
  • Author(s): 小澤龍彦, 岸裕幸, 村口篤
  • Publisher: 医薬の門社、感染・炎症・免疫
• [Book] 新しい抗体スクリーニング法 (2011)
  • Author(s): 小澤龍彦 他
  • Total Pages: 9
  • Publisher: 医薬の門社「感染・炎症・免疫」
• [Book] シングル細胞チップと抗体医薬への応用 (2011)
  • Author(s): 小澤龍彦 他
  • Total Pages: 8
  • Publisher: シーエムシー出版「ナノ融合による先進バイオデバイス」
• [Book] リンパ球アレイ法を用いた抗体医薬の開発 (2011)
  • Author(s): 小澤龍彦 他
  • Total Pages: 4
  • Publisher: ニューサイエンス社「細胞」
• [Patent(Industrial Property Rights)] 抗原特異的ウサギ抗体産生細胞の迅速な特定方法およびその利用 (2012)
  • Inventor(s): 村口篤, 岸裕幸, 小澤龍彦
  • Industrial Property Rights Holder: 村口篤, 岸裕幸, 小澤龍彦
  • Industrial Property Number: 2012-221822
  • Filing Date: 2012-10-04