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Study of sensitive immunosensor with the ZnO-coated plasmonic chip

Research Project

Project/Area Number 25286032
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypePartial Multi-year Fund
Section一般
Research Field Nanobioscience
Research InstitutionKwansei Gakuin University (2016)
National Institute of Advanced Industrial Science and Technology (2013-2015)

Principal Investigator

TAWA Keiko  関西学院大学, 理工学部, 教授 (80344109)

Co-Investigator(Kenkyū-buntansha) 梅津 光央  東北大学, 工学(系)研究科(研究院), 教授 (70333846)
Project Period (FY) 2013-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥18,850,000 (Direct Cost: ¥14,500,000、Indirect Cost: ¥4,350,000)
Fiscal Year 2015: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2014: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2013: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
Keywordsプラズモン / 格子 / サンドイッチアッセイ / IL-6 / 蛍光 / 酸化亜鉛 / プラズモン共鳴 / 増強蛍光 / バイオセンサー / 二重特性抗体 / イムノアッセイ / 二重特異性抗体 / 高感度検出 / イムノセンサー / 表面プラズモン共鳴
Outline of Final Research Achievements

Interleukin-6(IL-6), a cytokine relating inframmatory and autoimmune activity, was detected with fluorescence assays using a plasmonic chip. In their assays, the way of surface modification, sample volume, incubation time and mixing solution, were found to influence the detection sensitivity. When the assay was revised in the point of a rapid and easy process, the detection sensitivity was not compromised compared to assays with sufficient sample volume and assay
time. In the assay with the ZnO-plasmonic chip and bispecific antibody, the limit of detection was found to be pM level for 10 microL of sample solution and 20 min of assay time.

Report

(5 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Annual Research Report
  • 2014 Annual Research Report
  • 2013 Annual Research Report
  • Research Products

    (48 results)

All 2017 2016 2015 2014 2013 Other

All Journal Article (14 results) (of which Peer Reviewed: 14 results,  Open Access: 2 results,  Acknowledgement Compliant: 1 results) Presentation (32 results) (of which Int'l Joint Research: 12 results,  Invited: 8 results) Book (1 results) Remarks (1 results)

  • [Journal Article] Polydopamine Thin Films as Protein Linker Layer for Sensitive Detection of Interleukin-6 by Surface Plasmon Enhanced Fluorescence Spectroscopy2016

    • Author(s)
      Mana Toma and Keiko Tawa*
    • Journal Title

      ACS Appl. Mater. & Interfaces

      Volume: 8 Issue: 34 Pages: 22032-22038

    • DOI

      10.1021/acsami.6b06917

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Surface plasmon-enhanced optical trapping of quantum-dot-conjugated surface molecules on neurons cultured on a plasmonic chip2016

    • Author(s)
      K. Miyauchi, K. Tawa, Suguru N. Kudoh, T. Taguchi and C. Hosokawa
    • Journal Title

      Japanese Journal of Applied Physics

      Volume: 55 Issue: 6S1 Pages: 06GN04-06GN04

    • DOI

      10.7567/jjap.55.06gn04

    • NAID

      210000146677

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Sensitive Detection of Cell Surface Membrane Proteins in Living Breast Cancer Cells by Using Multicolor Fluorescence Microscopy with a Plasmonic Chip2016

    • Author(s)
      Keiko Tawa*, Shohei Yamamura, Chisato Sasakawa, Izumi Shibata, Masatoshi Kataoka
    • Journal Title

      ACS Appl. Mater. & Interfaces

      Volume: 8 Issue: 44 Pages: 29893-29898

    • DOI

      10.1021/acsami.6b07777

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Interleukin-6 Detection with a Plasmonic Chip2016

    • Author(s)
      Keiko Tawa*, Masashi Sumiya, Mana Toma, Chisato Sasakawa, Takuma Sujino, Tatsuki Miyaki, Hikaru Nakazawa, Mitsuo Umetsu
    • Journal Title

      Journal of Molecular and Engineering Materials

      Volume: 4 Issue: 03 Pages: 1640009-1640009

    • DOI

      10.1142/s2251237316400098

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A plasmonic chip-based bio/chemical hybrid sensing system for the highly sensitive detection of C-reactive protein2016

    • Author(s)
      Matsuura, R,, Tawa, K., Kitayama, Y., Takeuchi, T.
    • Journal Title

      Chemical Communications

      Volume: 52 Issue: 20 Pages: 3883-3886

    • DOI

      10.1039/c5cc07868g

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Journal Article] Fluorescence Microscopic Imaging of Cells with a Plasmonic Dish Integrally Molded2016

    • Author(s)
      Keiko Tawa, Chisato Sasakawa, Tsuyoshi Fujita, Kazuyuki Kiyosue, Chie Hosokawa, Junji Nishii, Makoto Oike, and Norihiro Kakinuma
    • Journal Title

      Japanese Journal of Applied Physics

      Volume: 55

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Surface plasmon-enhanced optical trapping of cell surface molecules on neurons cultured onto a plasmonic chip2016

    • Author(s)
      Kohei Miyauchi, Keiko Tawa, Suguru N. Kudoh, Takahisa Taguchi, and Chie Hosokawa
    • Journal Title

      Japanese Journal of Applied Physics

      Volume: 56

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Sensitive Detection of a Tumor Marker, α-Fetoprotein, with a Sandwich Assay on a Plasmonic Chip2015

    • Author(s)
      Keiko Tawa, Fusanori Kondo, Chisato Sasakawa, Kousuke Nagae, Yukito Nakamura, Akitoshi Nozaki, Takatoshi Kaya
    • Journal Title

      Analytical Chemistry

      Volume: 87 Issue: 7 Pages: 3871-3876

    • DOI

      10.1021/ac504642j

    • Related Report
      2015 Annual Research Report 2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] In Situ Sensitive Fluorescence Imaging of Neurons Cultured on a Plasmonic Dish Using Fluorescence Microscopy2014

    • Author(s)
      K. Tawa*, C. Yasui, C. Hosokawa, H. Aota and J. Nishii
    • Journal Title

      ACS Appl. Mater. Interfaces

      Volume: 6 Issue: 22 Pages: 20010-20015

    • DOI

      10.1021/am505579u

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Sensitive detection of interleukin-6 (IL-6) on a plasmonic chip by grating-coupled surface-plasmon-field-enhanced fluorescence imaging2014

    • Author(s)
      M. Tsuneyasu, C. Sasakawa,N. Naruishi, Y. Tanaka, Y. Yoshida, K. Tawa*,
    • Journal Title

      Japanese Journal of Applied Physics

      Volume: 53 Issue: 6S Pages: 06JL05-06JL05

    • DOI

      10.7567/jjap.53.06jl05

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Detection of Brain-Derived Neurotrophic Factor (BDNF) with a Sandwich Assay on a Plasmonic Chip2014

    • Author(s)
      Mari Satoh, Keiko Tawa*, Koichi Uegaki, Tomoko Hara, Mitsuo Umetsu, Hikaru Nakazawa, Makoto Itakura, Masami Takahashi, Hiroyuki Aota, and Masami Kojima
    • Journal Title

      Transaction of the Material Research Society of Japan

      Volume: 39 Pages: 361-364

    • NAID

      130005004361

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Sensitive detection of interleukin-6 (IL-6) on a plasmonic chip by grating-coupled surface-plasmon-field-enhanced fluorescence imaging2014

    • Author(s)
      M. Tsuneyasu, C. Sasakawa, N. Naruishi, Y. Tanaka, Y. Yoshida, K. Tawa
    • Journal Title

      Jap. J. Appl. Phys.

      Volume: 53

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Application of 300x Enhanced Fluorescence on a Plasmonic Chip Modified with a Bispecific Antibody to a Sensitive Immunosensor2013

    • Author(s)
      Keiko Tawa, Mitsuo Umetsu, Hikaru Nakazawa, Takamitsu Hattori, Izumi Kumagai
    • Journal Title

      ACS Applied Materials & Interfaces

      Volume: 5 Issue: 17 Pages: 8628-8632

    • DOI

      10.1021/am402173y

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Rapid and Sensitive Detection of Brain-Derived Neurotrophic Factor with a Plasmonic Chip2013

    • Author(s)
      Keiko Tawa, Mari Satoh, Koichi Uegaki, Tomoko Hara, Masami Kojima, Hiroyuki Aota, Yoshiki Yokota, Takahiko Nakaoki, Mitsuo Umetsu, Hikaru Nakazawa, Izumi Kumagai
    • Journal Title

      Japanese Journal of Applied Physics

      Volume: 52 Issue: 6S Pages: 06GK01-06GK01

    • DOI

      10.7567/jjap.52.06gk01

    • NAID

      210000142391

    • Related Report
      2013 Annual Research Report
    • Peer Reviewed
  • [Presentation] アルミニウムチップ上に調製されたジアリールエテン薄膜の光異性化に伴う結晶化過程のin situ 顕微鏡観察2017

    • Author(s)
      加登山太河、西村涼、當麻真奈、内田欣吾、田和圭子
    • Organizer
      日本化学会年会
    • Place of Presentation
      慶應義塾大学日吉キャンパス(神奈川県、横浜市)
    • Year and Date
      2017-03-17
    • Related Report
      2016 Annual Research Report
  • [Presentation] Bull’s Eye構造のプラズモニックチップ上における多重染色された乳癌細胞の蛍光顕微鏡観察2017

    • Author(s)
      泉 章太, 山村 昌平, 林 尚子, 當麻 真奈, 田和 圭子
    • Organizer
      応用物理学会春季学術講演会
    • Place of Presentation
      パシフィコ横浜(神奈川県、横浜市)
    • Year and Date
      2017-03-14
    • Related Report
      2016 Annual Research Report
  • [Presentation] Application of the Plasmonic Chip to Fluorescence Imaging and Photochemistry of a Diarylethene Film2016

    • Author(s)
      K. Tawa
    • Organizer
      9th Asian and Oceanian Photochemistry Conference 2016 (APC2016)
    • Place of Presentation
      Singapore
    • Year and Date
      2016-12-04
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] In-situ microspectroscopic imaging of photoisomerization and crystallization in diarylethene film on the aluminium plasmonic chip2016

    • Author(s)
      Taiga Kadoyama, Ryo Nishimura, Mana Toma, Kingo Uchida, Keiko Tawa
    • Organizer
      PhoSM2016
    • Place of Presentation
      名古屋大学(愛知県、名古屋市)
    • Year and Date
      2016-11-22
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Rapid and Sensitive detection of interleukin-6 with a sandwich immunoassay on the plasmonic chip2016

    • Author(s)
      K. Tawa, M. Sumiya, C. Sasakawa, T. Sujino, H. Nakazawa, and M. Umetsu
    • Organizer
      MNC2016
    • Place of Presentation
      ANAクラウンプラザホテル(京都府、京都市)
    • Year and Date
      2016-11-09
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Application of a Bull’s Eye-plasmonic Chip to Highly Sensitive Biodetection with a Microscop2016

    • Author(s)
      Shota Izumi, Mana Toma, Chie Hosokawa, Keiko Tawa
    • Organizer
      MNC2016
    • Place of Presentation
      ANAクラウンプラザホテル(京都府、京都市)
    • Year and Date
      2016-11-09
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 顕微鏡の照射角制御によって最適化されたプラズモニックチップ上のナノ粒子の蛍光増強度2016

    • Author(s)
      泉 章太,當麻 真奈,細川 千絵,田和 圭子
    • Organizer
      応用物理学会秋季学術講演会
    • Place of Presentation
      朱鷺メッセ(新潟県、新潟市)
    • Year and Date
      2016-09-13
    • Related Report
      2016 Annual Research Report
  • [Presentation] Multicolor Fluorescence imaging of living tumor cells with the plasmonic chip2016

    • Author(s)
      K. Tawa, C. Sasakawa, I.Shibata, S. Yamamura, and M. Kataoka
    • Organizer
      NFO-14
    • Place of Presentation
      浜松アクトシティ(静岡県、浜松市)
    • Year and Date
      2016-09-04
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] SENSITIVE IMMUNOSENSOR WITH A PLASMONIC CHIP PROVIDING THE ENHANCED FLUORESCENCE2016

    • Author(s)
      K. Tawa, C. Sasakawa, T. Sujino, M. Umetsu
    • Organizer
      IUPAC2016 International conference on Photochemstry
    • Place of Presentation
      大阪中央公会堂(大阪府、大阪市)
    • Year and Date
      2016-04-03
    • Related Report
      2016 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 酸化亜鉛プラズモニックチップを用いたサンドイッチ型高感度イムノセンシング2016

    • Author(s)
      田和圭子、角谷真詩、笹川知里、筋野拓馬、中澤光、梅津光央
    • Organizer
      第63回応用物理学会春季学術講演会
    • Place of Presentation
      東京工業大学大岡山キャンパス(東京都目黒区)
    • Year and Date
      2016-03-19
    • Related Report
      2015 Annual Research Report
  • [Presentation] Fluorescence imaging of a single nanoparticle on the plasmonic chip2015

    • Author(s)
      Keiko Tawa, Chisato Sasakawa, Chie Hosokawa
    • Organizer
      Pacifichem2015
    • Place of Presentation
      Honolulu, Hawaii (USA)
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Recombinant small antibody design for plasmonic biosensor using camel antibody with affinity for target inorganic material2015

    • Author(s)
      Takuma Sujino, Hikaru Nakazawa, Keiko Tawa, Ryutaro Asano, Izumi Kumagai, Mitsuo Umetsu
    • Organizer
      Pacifichem2015
    • Place of Presentation
      Honolulu, Hawaii (USA)
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Simple Detection of Interleukin-6 on a ZnO-Coated Plasmonic Chip with a Fluorescence Microscope2015

    • Author(s)
      K. Tawa, C. Sasakawa, T. Sujino, M. Umetsu
    • Organizer
      MNC2015
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2015-11-11
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Sensitive Fluorescence Imaging of Cells with a Palsmonic Chip2015

    • Author(s)
      Keiko TAWA
    • Organizer
      2015 KJF International Conference on Organic Materials for Electronics and Photonics
    • Place of Presentation
      KAL hotel, Jeju island (Korea)
    • Year and Date
      2015-09-06
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research / Invited
  • [Presentation] Multicolor fluorescence microscopic imaging of cancer cells on the plasmonic chip2015

    • Author(s)
      Keiko Tawa, Chisato Sasakawa, Shohei Yamamura, Izumi Shibata, Masatoshi Kataoka
    • Organizer
      SPIE Optics + Photonics, Nanoscience + Engineering, Plasmonics
    • Place of Presentation
      Convention Center, San Diego (USA)
    • Year and Date
      2015-08-09
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Fluorescence imaging of cells on the plasmonic dish integrally molded2015

    • Author(s)
      Keiko Tawa, Tsuyoshi Fujita, Chisato Sasakawa, Kazuyuki Kiyosue, Chie Hosokawa, Junji Nishii, Norihiro Kakinuma, Makoto Oike
    • Organizer
      第8回有機分子・バイオエレクトロニクス国際会議(M&BE8)
    • Place of Presentation
      タワーホール船堀(東京都江戸川区)
    • Year and Date
      2015-06-22
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] プラズモニックディッシュで培養した神経細胞の高感度蛍光イメージング2015

    • Author(s)
      田和圭子、藤田剛、細川千絵、西井準治
    • Organizer
      第64回高分子学会年次大会
    • Place of Presentation
      札幌コンベンションセンター(北海道札幌市)
    • Year and Date
      2015-05-27
    • Related Report
      2015 Annual Research Report
  • [Presentation] プラズモニックチップを用いた高感度蛍光検出法のバイオイメージングとバイオセンシングへの応2015

    • Author(s)
      田和圭子
    • Organizer
      応用物理学会春季学術講演会
    • Place of Presentation
      東海大(神奈川)
    • Year and Date
      2015-03-13
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] プラズモニックチップを用いた高感度イムノセンシングとバイオイメージング2014

    • Author(s)
      田和圭子
    • Organizer
      分光学会関西支部講演会
    • Place of Presentation
      大阪電気通信大学(大阪)
    • Year and Date
      2014-11-28
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 格子結合型プラズモン増強蛍光を利用したバイオセンシングとバイオイメージング2014

    • Author(s)
      田和圭子
    • Organizer
      第7回プラズモニック化学シンポジウム
    • Place of Presentation
      首都大(東京)
    • Year and Date
      2014-11-14
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] Multicolor fluorescence microscopic imaging of tumor cells enhanced with the plasmonic chip2014

    • Author(s)
      田和 圭子、笹川 知里、芝田 いずみ、山村 昌平、片岡 正俊
    • Organizer
      MNC2014
    • Place of Presentation
      ヒルトン福岡シーホーク(福岡)
    • Year and Date
      2014-11-06
    • Related Report
      2014 Annual Research Report
  • [Presentation] Sensitive detection of α-fetoprotein (AFP) with the enhanced fluorescence on the plasmonic chip2014

    • Author(s)
      田和 圭子、近藤 房宣、永江剛典、中村幸登、彼谷隆敏
    • Organizer
      IUMRS-ICA2014
    • Place of Presentation
      福岡大学(福岡)
    • Year and Date
      2014-08-26
    • Related Report
      2014 Annual Research Report
  • [Presentation] Bright fluorescence imaging of cells cultured on the plasmonic dish2014

    • Author(s)
      田和 圭子、藤田 剛、安井力、清末 和之、細川 千絵、笹川 知里、青田浩幸、柿沼憲宏、尾池真、西井準治
    • Organizer
      the 248th ACS National Meeting
    • Place of Presentation
      Moscone Center (San Francisco, USA)
    • Year and Date
      2014-08-12
    • Related Report
      2014 Annual Research Report
  • [Presentation] プラズモニックチップを用いた生体分子の高感度検出2014

    • Author(s)
      田和圭子
    • Organizer
      第204回有機エレクトロニクス材料研究会
    • Place of Presentation
      新宿NSビル(東京)
    • Year and Date
      2014-04-24
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] 表面プラズモン励起増強蛍光イメージングによるプラズモニックチップ上での生活習慣病マーカーIL-6の高感度マルチアレイ計測

    • Author(s)
      常安 将央、笹川 知里、鳴石 奈穂子、田中喜秀学、吉田 康一、田和 圭子
    • Organizer
      応用物理学会春季講演会
    • Place of Presentation
      青山学院大学(神奈川県)
    • Related Report
      2013 Annual Research Report
  • [Presentation] プラズモニックチップによる高感度イムノセンシング

    • Author(s)
      田和圭子
    • Organizer
      応用物理学会春季講演会
    • Place of Presentation
      青山学院大学(神奈川県)
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Presentation] Detection of Brain-Derived Neurotrophic Factor (BDNF) on the ZnO-coated plasmonic chip

    • Author(s)
      M. Satoh, K. Tawa, K. Uegaki, T. Hara, M. Umetsu, H. Nakazawa, M. Itakura, M. Takahashi, H. Aota, M. Kojima
    • Organizer
      MRS-Japan
    • Place of Presentation
      横浜
    • Related Report
      2013 Annual Research Report
  • [Presentation] Sensitive and multi detection of Interleukin-6 (IL-6) by Grating Coupled-Surface Plasmon-field enhanced Fluorescence Imaging

    • Author(s)
      M. Tsuneyasu, C. Sasakawa, N. Naruishi, Y. Tanaka, Y. Yoshida, K. Tawa
    • Organizer
      MNC2013
    • Place of Presentation
      札幌
    • Related Report
      2013 Annual Research Report
  • [Presentation] Sensitive detection of Interleukin-6 (IL-6) on the plasmonic chip by Grating Coupled-Surface Plasmon-field enhanced Fluorescence Imaging

    • Author(s)
      M. Tsuneyasu, C. Sasakawa, N. Naruishi, Y. Tanaka, Y. Yoshida, K. Tawa
    • Organizer
      JSAP-OSA Joint Symposium
    • Place of Presentation
      同志社大学(京都)
    • Related Report
      2013 Annual Research Report
  • [Presentation] 酸化亜鉛コーティングプラズモニックチップ上のサンドイッチアッセイによるうつ病関連マーカーBDNFの迅速検出

    • Author(s)
      M. Satoh, K. Tawa, K. Uegaki, T. Hara, M. Umetsu, H. Nakazawa, I. Kumagai, M. Itakura, M. Takahashi, H. Aota, M. Kojima
    • Organizer
      高分子学会年次大会
    • Place of Presentation
      京都国際会館
    • Related Report
      2013 Annual Research Report
  • [Presentation] 材料標的抗体を錨としたバイオセンサー指向インターフェイス設計

    • Author(s)
      梅津光央、田和 圭子、中澤光、筋野 拓馬、服部 峰充
    • Organizer
      化学工学会 第45回秋季大会
    • Place of Presentation
      岡山大学 (岡山)
    • Related Report
      2013 Annual Research Report
  • [Presentation] ナノを意識した蛋白質スマートデザイン

    • Author(s)
      梅津 光央
    • Organizer
      生体機能関連化学部会 第28回若手フォーラム
    • Place of Presentation
      名古屋大学(名古屋)
    • Related Report
      2013 Annual Research Report
    • Invited
  • [Book] プラズモンナノ材料開発の最前線と応用(第7章 格子結合型表面プラズモン励起増強蛍光(GC-SPF)法を用いた生体分子検出)2013

    • Author(s)
      田和圭子
    • Total Pages
      8
    • Publisher
      シーエムシー出版
    • Related Report
      2013 Annual Research Report
  • [Remarks] http://www.kg-applchem.jp/tawa/

    • Related Report
      2016 Annual Research Report

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Published: 2013-05-21   Modified: 2019-07-29  

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