糖タンパク質を標的とした革新的がん特異的抗体の開発とその作用機序の解明

2021 Fiscal Year Annual Research Report

• Project/Area Number: 19K07705
• Research Institution: Tohoku University
• Principal Investigator: 加藤 幸成 東北大学, 医学系研究科, 教授 (00571811)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: 抗体 / モノクローナル抗体 / がん特異的抗体 / エピトープ / エピトープマッピング / CasMab法 / 抗体医薬 / がん

Outline of Annual Research Achievements

モノクローナル抗体は、がんを中心としたあらゆる病気の診断や治療に活用されている。がん細胞を狙う場合、どんなに抗体を改変しても、あるいは、どんなに強力な抗がん剤付加や免疫細胞誘導を行ったとしても、がん細胞に特異的な抗体でなければ、常に正常組織への毒性が懸念される。しかし、がん細胞のみに高発現している分子は限られており、理想的な標的分子は枯渇している。我々が開発したがん特異的抗体作製法（CasMab法）では、がん特異的糖鎖が付加した糖タンパク質や細胞株を免疫することにより、がん特異的抗体を直接取得することができる。本研究課題では、がん細胞、正常細胞で共に発現しているタンパク質を標的とする。既に抗体医薬が開発されているが、より優れた抗体が求められるもの、及び抗体医薬が存在しない新規のものを標的分子とする。具体的には、３種類の標的タンパク質に対して行った。これまで、マウスに標的タンパク質発現細胞株あるいは分泌型の標的タンパク質を免疫することにより、３種類の標的分子に対する抗体のライブラリーを作製した。抗体のスクリーニングは、精製タンパク質や標的タンパク質発現細胞を用いて行った。細胞株を免疫する場合は、親株と発現株に対する反応性の違いを、フローサイトメトリー（研究室所有の96well全自動アナライザー）で検出した。さらに、３種類の標的タンパク質に対して作製した各種クローンについて、フローサイトメトリー、ウェスタンブロット、免疫組織染色により活性評価を行なった。令和3年度には、樹立および性状解析をした各種抗体のエピトープ解析を行った。具体的には、３種類の標的タンパク質について、欠損変異体、点突然変異体を作製し、エピトープ解析を実施した。今後、これらの抗体の臨床応用について検討していく。

Research Products

• [Journal Article] Epitope Mapping of an Anti-CD20 Monoclonal Antibody (C20Mab-60) using Enzyme-Linked Immunosorbent Assay (2021)
  • Author(s): Takei J, Suzuki H, Asano T, Li G, Saito M, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 250-254
  • DOI: 10.1089/mab.2021.0042
  • Peer Reviewed
• [Journal Article] Epitope Mapping of an Anti-Human CCR9 Monoclonal Antibody (C9Mab-1) using Enzyme-Linked Immunosorbent Assay (2021)
  • Author(s): Takei J, Asano T, Li G, Saito M, Suzuki H, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 239-242
  • DOI: 10.1089/mab.2021.0037
  • Peer Reviewed
• [Journal Article] Epitope Mapping of an Anti-HER2 Monoclonal Antibody (H2Mab-181) using Enzyme-Linked Immunosorbent Assay (2021)
  • Author(s): Asano T, Takei J, Suzuki H, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 255-260
  • DOI: 10.1089/mab.2021.0029
  • Peer Reviewed
• [Journal Article] Epitope Mapping of an Anti-CD20 Monoclonal Antibody (C20Mab-60) using the HisMAP Method (2021)
  • Author(s): Asano T, Takei J, Furusawa Y, Saito M, Suzuki H, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 243-249
  • DOI: 10.1089/mab.2021.0035
  • Peer Reviewed
• [Journal Article] Defucosylated Mouse-Dog Chimeric Anti-EGFR Antibody Exerts Antitumor Activities in Mouse Xenograft Models of Canine Tumors (2021)
  • Author(s): Li G, Ohishi T, Kaneko MK, Takei J, Mizuno T, Kawada M, Saito M, Suzuki H, Kato Y.
  • Journal Title: Cells Volume: 10 Pages: 3599
  • DOI: 10.3390/cells10123599
  • Peer Reviewed / Open Access
• [Journal Article] Epitope mapping of the anti-CD44 monoclonal antibody (C44Mab-46) using alanine-scanning mutagenesis and surface plasmon resonance (2021)
  • Author(s): Takei J, Asano T, Suzuki H, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 219-226
  • DOI: 10.1089/mab.2021.0028
  • Peer Reviewed
• [Journal Article] Preclinical evaluation of podoplanin-targeted al-pha-radioimmunotherapy with the novel antibody NZ-16 for malignant mesothelioma (2021)
  • Author(s): Asano T, Suzuki H, Kaneko MK, Kato Y.
  • Journal Title: Cells Volume: 10 Pages: 2503
  • DOI: 10.3390/cells10102503
  • Peer Reviewed / Open Access
• [Journal Article] An Anti‐HER2 Monoclonal Antibody H2Mab‐41 Exerts Antitumor Activities in Mouse Xenograft Model using Dog HER2‐overexpressed Cells (2021)
  • Author(s): Tateyama N, Asano T, Ohishi T, Takei J, Hosono H, Nanamiya R, Tanaka T, Sano M, Saito M, Kawada M, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 184-190
  • DOI: 10.1089/mab.2021.0025
  • Peer Reviewed
• [Journal Article] Defucosylated Anti-EGFR Monoclonal Antibody 134-mG2a-f Exerts Antitumor Activities in Mouse Xenograft Models of Dog EGFR-overexpressed Cells (2021)
  • Author(s): Tateyama N, Nanamiya R, Ohishi T, Takei J, Nakamura T, Yanaka M, Hosono H, Saito M, Asano T, Tanaka T, Sano M, Kawada M, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 177-183
  • DOI: 10.1089/mab.2021.0022
  • Peer Reviewed
• [Journal Article] Development of a novel epitope mapping system: RIEDL insertion for epitope mapping method (2021)
  • Author(s): Asano T, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 162-167
  • DOI: 10.1089/mab.2021.0023
  • Peer Reviewed
• [Journal Article] Epitope Mapping of an Anti-Human EGFR Monoclonal Antibody (EMab-51) using the REMAP Method (2021)
  • Author(s): Nanamiya R, Sano M, Asano T, Yanaka M, Nakamura T, Saito M, Tanaka T, Hosono H, Tateyama N, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 149-155
  • DOI: 10.1089/mab.2021.0010
  • Peer Reviewed
• [Journal Article] Development of a Novel Anti-HER2 Monoclonal Antibody H2Mab-181 for Gastric Cancer (2021)
  • Author(s): Takei J, Asano T, Tanaka T, Sano M, Hosono H, Nanamiya R, Tateyama N, Saito M, Suzuki H, Harada H, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 168-176
  • DOI: 10.1089/mab.2021.0021
  • Peer Reviewed
• [Journal Article] Epitope Mapping of the Anti-CD44 Monoclonal Antibody (C44Mab-46) Using the REMAP Method (2021)
  • Author(s): Asano T, Kaneko MK, Takei J, Tateyama N, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 156-161
  • DOI: 10.1089/mab.2021.0012
  • Peer Reviewed
• [Journal Article] Epitope Mapping of an Anti-human EGFR Monoclonal Antibody (EMab-134) using the REMAP Method (2021)
  • Author(s): Sano M, Kaneko MK, Asano T, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 191-195
  • DOI: 10.1089/mab.2021.0014
  • Peer Reviewed
• [Journal Article] Epitope Mapping of the Anti-California Sea Lion Podoplanin Monoclonal Antibody PMab-269 using Alanine-scanning Mutagenesis and ELISA (2021)
  • Author(s): Tanaka T, Asano T, Sano M, Takei J, Hosono H, Nanamiya R, Tateyama N, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 196-200
  • DOI: 10.1089/mab.2021.0017
  • Peer Reviewed
• [Journal Article] Anti-HER3 monoclonal antibody exerts antitumor activity in a mouse model of colorectal adenocarcinoma (2021)
  • Author(s): Asano T, Ohishi T, Takei J, Nakamura T, Nanamiya R, Hosono H, Tanaka T, Sano M, Harada H, Kawada M, Kaneko MK, Kato Y.
  • Journal Title: Oncol. Rep Volume: 46 Pages: 173
  • DOI: 10.3892/or.2021.8124
  • Peer Reviewed / Open Access
• [Journal Article] Ferret Podoplanin is Detected by PMab-241 in Immunohistochemistry (2021)
  • Author(s): Nanamiya R, Takei J, Asano T, Sano M, Tanaka T, Hosono H, Harada H, Sakai Y, Mizuno T, Suzuki H, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 134-140
  • DOI: 10.1089/mab.2021.0016
  • Peer Reviewed
• [Journal Article] Development of Anti-Human CC Chemokine Receptor 9 Monoclonal Antibodies for Flow Cytometry (2021)
  • Author(s): Nanamiya R, Takei J, Asano T, Tanaka T, Sano M, Nakamura T, Yanaka M, Hosono H, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 101-106
  • DOI: 10.1089/mab.2021.0007
  • Peer Reviewed
• [Journal Article] Development of an Anti-Elephant Podoplanin Monoclonal Antibody PMab-265 for Flow cytometry (2021)
  • Author(s): Hosono H, Asano T, Takei J, Sano M, Tanaka T, Kaneko MK, Kato Y
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 141-145
  • DOI: 10.1089/mab.2021.0015
  • Peer Reviewed
• [Journal Article] Development of Monoclonal Antibody PMab-269 Against California Sea Lion Podoplanin (2021)
  • Author(s): Tanaka T, Asano T, Sano M, Takei J, Hosono H, Nanamiya R, Nakamura T, Yanaka M, Harada H, Fukui M, Suzuki H, Uchida K, Nakagawa T, Kato Y, Kaneko MK.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 124-133
  • DOI: 10.1089/mab.2021.0011
  • Peer Reviewed
• [Journal Article] Development of Anti-Mouse CC Chemokine Receptor 3 Monoclonal Antibodies for Flow Cytometry (2021)
  • Author(s): Asano A, Nanamiya R, Takei J, Nakamura T, Yanaka M, Hosono H, Tanaka T, Sano M, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 107-112
  • DOI: 10.1089/mab.2021.0009
  • Peer Reviewed
• [Journal Article] An anti-TROP2 monoclonal antibody TrMab-6 exerts antitumor activity in breast cancer mouse xenograft models (2021)
  • Author(s): Tanaka T, Ohishi T, Asano T, Takei J, Nanamiya R, Hosono H, Sano M, Harada H, Kawada M, Kaneko MK, Kato Y.
  • Journal Title: Oncol Rep Volume: 46 Pages: 132
  • DOI: 10.3892/or.2021.8083
  • Peer Reviewed / Open Access
• [Journal Article] Development of Anti-Mouse CC chemokine receptor 8 Monoclonal Antibodies for Flow Cytometry (2021)
  • Author(s): Tanaka T, Nanamiya R, Takei J, Nakamura T, Yanaka M, Hosono H, Sano M, Asano T, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 65-70
  • DOI: 10.1089/mab.2021.0005
• [Journal Article] Development of anti-human killer cell lectin-like receptor subfamily G member 1 monoclonal antibodies for flow cytometry (2021)
  • Author(s): Asano T, Nanamiya R, Tanaka T, Kaneko MK, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 76-80
  • DOI: 10.1089/mab.2021.0008
  • Peer Reviewed
• [Journal Article] Development of Anti-Human T-Cell Immunoreceptor with Ig and ITIM Domains (TIGIT) Monoclonal Antibodies for Flow Cytometry (2021)
  • Author(s): Takei J, Asano T, Nanamiya R, Nakamura T, Yanaka M, Hosono H, Tanaka T, Sano M, Kaneko MK, Harada H, Kato Y.
  • Journal Title: Monoclon. Antib. Immunodiagn. Immunother. Volume: 40 Pages: 71-75
  • DOI: 10.1089/mab.2021.0006
  • Peer Reviewed
• [Journal Article] Establishment of a novel anti-TROP2 monoclonal antibody TrMab-29 for immunohistochemical analysis (2021)
  • Author(s): Sayama Y, Kaneko MK, Takei J, Hosono H, Sano M, Asano T, Kato Y.
  • Journal Title: Biochem Biophys Rep Volume: 25 Pages: 100902
  • DOI: 10.1016/j.bbrep.2020.100902
  • Peer Reviewed / Open Access
• [Journal Article] Development and characterization of TrMab-6, a novel anti-TROP2 monoclonal antibody for antigen detection in breast cancer (2021)
  • Author(s): Sayama Y, Kaneko MK, Kato Y.
  • Journal Title: Mol Med Rep Volume: 23 Pages: 92
  • DOI: 10.3892/mmr.2020.11731
  • Peer Reviewed / Open Access
• [Presentation] ペプチドタグシステムを用いた新規抗体エピトープ解析法の開発 (2021)
  • Author(s): 加藤幸成、淺野禎三、金子美華
  • Organizer: 第21回 日本蛋白質科学会年会
  • Invited
• [Presentation] ヒト抗体医薬の現状と未来 (2021)
  • Author(s): 加藤幸成
  • Organizer: 第12回動物用ワクチン・バイオ医薬研究会シンポジウム『動物用バイオ医薬品の最先端：動物用抗体医薬の現状と未来』
  • Invited
• [Remarks] 東北大学大学院医学系研究科分子薬理学分野・業績
  • URL: http://www.med-tohoku-antibody.com/reference/01.htm