Analysis on superactive clustering structure of recombinant antibodies and study on its production process with high efficiency

• Project/Area Number: 19760548
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Biofunction/Bioprocess
• Research Institution: Tohoku University
• Principal Investigator: ASANO Ryutaro Tohoku University, 大学院・工学研究科, 助教 (80323103)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥3,690,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥390,000); Fiscal Year 2008: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2007: ¥2,000,000 (Direct Cost: ¥2,000,000)
• Keywords: 二重特異性抗体 / 二重特異性diabody / 多量体化 / クラスター構造 / 多価化 / がん細胞傷害活性 / 表面プラズモン共鳴法 / 等温滴定型熱量測定 / diabody / EGFR / フローサイトメトリー / scFv / 二重特性抗体 / 多価抗体 / 一本鎖抗体 / 表面プラズモン共鳴

Research Abstract

研究代表者らは、独自に開発した低分子組換え抗体の調製時に、微量だが強力ながん細胞殺傷能を有する高分子量の分子種が含まれることを、近年明らかにした。本研究は、この構造体の詳細な解析と高効率な調製プロセスの開発を目的に進めた。各種手法を用いた精密解析の結果、高分子は比較的均一な四量体分子であることが明らかになり、またリンカー配列の改変や、大腸菌発現系の検討により、収量を向上させることにも成功した。

Research Products

• [Journal Article] 次世代抗体医薬の分子設計 (2009)
  • Author(s): 熊谷泉,浅野竜太郎
  • Journal Title: ケミカルエンジニヤリング 54 Pages: 65-71
• [Journal Article] 次世代抗体医薬の分子設計 (2009)
  • Author(s): 熊谷泉
  • Journal Title: ケミカルエンジニヤリング 54 Pages: 65-71
• [Journal Article] Diabody-based recombinant formats of humanized IgG-like bispecific antibody with effective retargeting of lymphocytes to tumor cells (2008)
  • Author(s): Asano R., Kawaguchi H., Watanabe Y., Nakanishi T., Umetsu M., Hayashi H., Katayose Y., Unno M., Kudo T., Kumagai I.
  • Journal Title: J. Immunother. 31 Pages: 752-761
  • Peer Reviewed
• [Journal Article] Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528 (2008)
  • Author(s): Asano R., Sone Y., Tkoma K., Hayashi H., Nakanishi T., Umetsu M., Katayose Y., Unno M., Kudo T., Kumagai I.
  • Journal Title: Protein Fng. Des. Sel. 10 Pages: 597-603
  • Peer Reviewed
• [Journal Article] Grafting of material-binding function into antibodies Functionalization by peptide grafting (2008)
  • Author(s): Hattori T., Umetsu M., Nakanishi T., Tsumoto K., Ohara S., Abe H., Naito M., Asano R., Adschiri T., Kumagai I.
  • Journal Title: Biochem. Biophys. Res. Commun. 365 Pages: 751-757
  • Peer Reviewed
• [Journal Article] Thermodynamic consequences of mutations in Vernier zone residues of a humanized anti-human epidermal growth factor receptor (EGFR) murine antibody, 528 (2008)
  • Author(s): Makabe K., Nakanishi T., Tsumoto K., Tanaka Y., Kondo H., Umetsu M., Sone Y., Asano R., Kumagai I.
  • Journal Title: J. Biol. Chem. 283 Pages: 1156-1166
  • Peer Reviewed
• [Journal Article] リンパ球とがん細胞を認識するヒト型化二重特異性抗体 (2008)
  • Author(s): 熊谷 泉,浅野竜太郎
  • Journal Title: Drug Delivery System 23 Pages: 518-525
• [Journal Article] Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528 (2008)
  • Author(s): Ryutaro Asano
  • Journal Title: Protein Engineering Design & Selection 10 Pages: 597-603
  • Peer Reviewed
• [Journal Article] Diabody-based recombinant formats of humanized IgG-like bispecific antibody with effective retargeting of lymphocytes to tumor cells (2008)
  • Author(s): Ryutaro Asano
  • Journal Title: Journal of Immunotherapy 31 Pages: 752-761
  • Peer Reviewed
• [Journal Article] リンパ球とがん細胞を認識するヒト型二重特異性抗体 (2008)
  • Author(s): 熊谷泉
  • Journal Title: Drug Delivery System 23 Pages: 518-525
• [Journal Article] Thermodynamic consequences of mutations in Vernier zone residues of a humanized anti-human epidermal growth factor receptor (EGFR) murine antibody, 528 (2008)
  • Author(s): Koki Makabe
  • Journal Title: The Journal of Biological Chemistry 283 Pages: 1156-1166
  • Peer Reviewed
• [Journal Article] Grafting of material-binding function into antibodies Functionalization by peptide grafting. (2008)
  • Author(s): Takamitsu Hattori
  • Journal Title: Biochemical and Biophysical Research Communications 365 Pages: 751-757
  • Peer Reviewed
• [Journal Article] High IL-21 receptor expression and apoptosis induction by IL-21 in follicular lymphoma (2007)
  • Author(s): Akamatsu N., Yamada Y., Hasegawa H., Makabe K., Asano R., Kumagai I., Murata K., Tmaizumi Y., Tsukasaki K,, Tsuruda K., Sugahara K., Atogami S., Yanagihara K., Kamihira S.
  • Journal Title: Cancer Lett. 256 Pages: 196-206
  • Peer Reviewed
• [Journal Article] Highly effective recombinant format of a humanized TgG-like bispecific antibody for cancer immunotherapy with retargeting of lymphocytes to tumor cells (2007)
  • Author(s): Asano R., Watanabe Y., Kawaguchi H., Fukazawa H., Nakanishi T., Umetsu M., Hayashi H., Katayose Y., Unno M., Kudo T., Kumagai I.
  • Journal Title: J. Biol. Chem. 282 Pages: 27659-27665
  • Peer Reviewed
• [Journal Article] Electrochemical screening of recombinant protein solubility in Escherichia coli using scanning electrochemical microscopy (SECM) (2007)
  • Author(s): Nagamine K., Onodera S., Kurihara A., Yasukawa T., Shiku H., Asano R., Kumagai T., Matsue T.
  • Journal Title: Biotechnol. Bioeng. 96 Pages: 1008-1013
  • Peer Reviewed
• [Journal Article] Highly effective recombinant format of a humanized IgG-like bispecific antibody for cancer immunotherapy with retargeting of lymphocytes to tumor cells. (2007)
  • Author(s): Ryutaro Asano
  • Journal Title: The Journal of Biological Chemistry 282 Pages: 27659-27665
  • Peer Reviewed
• [Journal Article] High IL-21 receptor expression and apoptosis induction by IL-21 in follicular lymphoma. (2007)
  • Author(s): Norihiko Akamatsu
  • Journal Title: Cancer Letters 256 Pages: 196-206
  • Peer Reviewed
• [Journal Article] Electrochemical screening of recombinant protein solubility in Escherichia coli using scanning electrochemical microscopy (SECM). (2007)
  • Author(s): Kuniaki Nagamine
  • Journal Title: Biotechnology and Bioengineering 96 Pages: 1008-1013
  • Peer Reviewed
• [Presentation] 低分子二重特異性抗体の新規調製法の開発 (2008)
  • Author(s): 浅野竜太郎
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会合同大会)
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-09
• [Presentation] 抗EGFR一本鎖抗体の多量体化とがん治療に向けた機能評価 (2008)
  • Author(s): 萩原康世
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会合同大会)
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-09
• [Presentation] 低分子抗体の多価化設計と物理化学的相互作用解析 (2008)
  • Author(s): 階上健太郎
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会合同大会)
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-09
• [Presentation] tandem scFv型二重特異性抗体の巻き戻しによる調製法の検討 (2008)
  • Author(s): 田中 翔
  • Organizer: BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会合同大会)
  • Place of Presentation: 神戸
  • Year and Date: 2008-12-09
• [Presentation] Efficacy of humanized IgG-like bispecific antibodies for cancer immunotherapy with EGER and CD3 retargeting (2008)
  • Author(s): 浅野竜太郎
  • Organizer: 第67回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-10-29
• [Presentation] Efficacy of humanized IgG-like bispecific antibodies for cancer immunot herapy with EGFR and CD3 retargeting (2008)
  • Author(s): 浅野竜太郎
  • Organizer: 第67回日本癌学会学術総会
  • Place of Presentation: 名古屋
  • Year and Date: 2008-10-29
• [Presentation] 動物細胞を用いた高機能性組換え型治療抗体の調製と機能評価 (2008)
  • Author(s): 浅野竜太郎
  • Organizer: 第60回日本生物工学会大会
  • Place of Presentation: 仙台
  • Year and Date: 2008-08-28
• [Presentation] 低分子二重特異性抗体の高活性クラスター構造解析 (2008)
  • Author(s): 浅野竜太郎
  • Organizer: 第8回日本蛋白質科学会年会
  • Place of Presentation: 東京
  • Year and Date: 2008-06-11
• [Presentation] EGFRとCD3を標的とした高機能な二重特異性抗体の形態 (2007)
  • Author(s): 浅野竜太郎
  • Organizer: BMB2007(第30回日本分子生物学会年会・第80回日本生化学会大会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-11
• [Presentation] ファージ提示法を用いた高親和性EGFR特異的抗体の選択 (2007)
  • Author(s): 田原和浩
  • Organizer: BMB2007(第30回日本分子生物学会年会・第80回日本生化学会大会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-11
• [Presentation] がん治療を目指したIgG様二重特異性抗体の高機能化 (2007)
  • Author(s): 石山優奈
  • Organizer: BMB2007(第30回日本分子生物学会年会・第80回日本生化学会大会合同大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-11
• [Presentation] EGFRとCD3を標的とした高機能な二重特異性抗体の形態 (2007)
  • Author(s): 浅野 竜太郎
  • Organizer: BMB2007(第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会)
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2007-12-11
• [Presentation] がん治療を目指したIgG様二重特異性抗体の高機能化 (2007)
  • Author(s): 石山 優奈
  • Organizer: BMB2007(第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会)
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2007-12-11
• [Presentation] ファージ提示法を用いた高親和性EGFR特異的抗体の選択 (2007)
  • Author(s): 田原 和浩
  • Organizer: BMB2007(第30回日本分子生物学会年会・第80回日本生化学会大会 合同大会)
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2007-12-11
• [Presentation] Efficacy of Humanized IgG-like Bispecific Antibody Formats in Cancer Immunotherapy (2007)
  • Author(s): Ryutaro Asano
  • Organizer: Antibodies - Europe
  • Place of Presentation: Wien
  • Year and Date: 2007-11-07
• [Presentation] Affinity Maturation of the Humanized Anti-EGFR Antibody Fragment by Phage Display (2007)
  • Author(s): Kazuhiro Tahara
  • Organizer: Antibodies - Europe
  • Place of Presentation: Wien
  • Year and Date: 2007-11-07
• [Presentation] Efficacy of Humanized IgG-like Bispecific Antibody Formats in Cancer Immunotherapy (2007)
  • Author(s): Ryutaro Asano
  • Organizer: Antibodies-Europe Engineering the Next Generation of Antibodies
  • Place of Presentation: InterContinental Wien
  • Year and Date: 2007-11-07
• [Presentation] Affinity Maturation of the Humanized Anti-EGFR Antibody Fragment by Phage Display (2007)
  • Author(s): Kazuhiro Tahara
  • Organizer: Antibodies-Europe Engineering the Next Generation of Antibodies
  • Place of Presentation: InterContinental Wien
  • Year and Date: 2007-11-07
• [Presentation] 生体分子の自己組織化能を利用した低分子抗体の高機能化 (2007)
  • Author(s): 渡邊志緒美
  • Organizer: 第22回生体機能関連化学シンポジウム
  • Place of Presentation: 仙台
  • Year and Date: 2007-09-29
• [Presentation] 生体分子の自己組織化能を利用した低分子抗体の高機能化 (2007)
  • Author(s): 渡邊志緒美
  • Organizer: 第22回生体機能関連化学シンポジウム
  • Place of Presentation: 東北大学多元物質科学研究所
  • Year and Date: 2007-09-29
• [Presentation] ジスルフィド安定化diabodyの構築に向けた検討 (2007)
  • Author(s): 萩原康世
  • Organizer: 第7回日本蛋白質科学会年会
  • Place of Presentation: 東京
  • Year and Date: 2007-05-26
• [Presentation] Tandem scFv型二重特異性抗体の大腸菌発現系を用いた調整と機能評価 (2007)
  • Author(s): 田中 翔
  • Organizer: 第7回日本蛋白質科学会年会
  • Place of Presentation: 東京
  • Year and Date: 2007-05-26
• [Presentation] ジスルフィド安定化diabodyの構築に向けた検討 (2007)
  • Author(s): 萩原 康世
  • Organizer: 第7回日本蛋白質科学会年会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2007-05-26
• [Presentation] Tandem scFv型二重特異性抗体の大腸菌発現系を用いた調整と機能評価 (2007)
  • Author(s): 田中 翔
  • Organizer: 第7回日本蛋白質科学会年会
  • Place of Presentation: 仙台国際センター
  • Year and Date: 2007-05-26
• [Book] 抗体ドメインの積み木細工による高機能化と医用への展開(抗体医薬の最前線) (2007)
  • Author(s): 熊谷泉,浅野竜太郎,中西猛
  • Publisher: シーエムシー出版
• [Book] 抗体ドメインの積み木細工による高機能化と医用への展開(抗体医薬の最前線) (2007)
  • Author(s): 熊谷 泉
  • Publisher: シーエムシー出版
• [Patent(Industrial Property Rights)] 多量体化低分子抗体 (2008)
  • Inventor(s): 熊谷 泉,浅野竜太郎,梅津光央
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 2008-292894
  • Filing Date: 2008-11-17
• [Patent(Industrial Property Rights)] 組換え型ポリペプチドの製造方法 (2008)
  • Inventor(s): 熊谷 泉,浅野竜太郎,中西 猛,梅津光央
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 2008-126612
  • Filing Date: 2008-05-14
• [Patent(Industrial Property Rights)] 組換え型ポリペプチドの製造方法 (2008)
  • Inventor(s): 熊谷泉, 浅野竜太郎, 中西猛, 梅津光央
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 2008-126612
  • Filing Date: 2008-05-14
• [Patent(Industrial Property Rights)] 多量体化低分子抗体 (2008)
  • Inventor(s): 熊谷泉, 浅野竜太郎, 梅津光央
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 2008-292894
  • Filing Date: 2008-11-17