Development of a method for fusion of biological cells onto supported lipid bilayers and its application to membrane protein analyses

• Project/Area Number: 26288067
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Analytical chemistry
• Research Institution: Okayama University
• Principal Investigator: KANETA Takashi 岡山大学, 自然科学研究科, 教授 (20243909)
• Co-Investigator(Renkei-kenkyūsha): SASAKI Keiko 九州大学, 大学院工学研究院, 教授 (30311525) ; TAKEYASU Nobuyuki 岡山大学, 大学院自然科学研究科, 准教授 (90373323)
• Research Collaborator: HIGASHIDANI Naoki ; KANAJI Keisuke ; KOBAYASHI Sakurako ; MAKI Tomomi ; OGAWA Kazuma ; KARITA Shingo ; MITSUNOBU Manami ; SHIMADA Yuhi ; HARADA Airi ; KUDO Simire ; ISOYAMA Mika ; KUBOI Mai ; FUJII Tatsuya ; MIKI Shoko
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥16,640,000 (Direct Cost: ¥12,800,000、Indirect Cost: ¥3,840,000); Fiscal Year 2016: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2015: ¥8,060,000 (Direct Cost: ¥6,200,000、Indirect Cost: ¥1,860,000); Fiscal Year 2014: ¥5,850,000 (Direct Cost: ¥4,500,000、Indirect Cost: ¥1,350,000)
• Keywords: 膜タンパク質 / 単一細胞計測 / 細胞融合 / 電気泳動 / 単一分子検出 / マイクロ流体デバイス

Outline of Final Research Achievements

The present study aimed the development of the method to incorporate membrane proteins in real biological cells into supported lipid bilayers and its application to the separation and determination of the membrane proteins. We constructed a system for observing total internal reflection fluorescence. We fabricated the supported lipid bilayers and incorporated biological cells into the supported lipid bilayers in order to develop the analytical method of the membrane proteins localized in biological cell membranes.

Research Products

• [Journal Article] Temperature-Induced Coalescence of Droplets Manipulated by Optical Trapping in an Oil-in-Water Emulsion (2017)
  • Author(s): Manami Mitsunobu, Sakurako Kobayashi, Nobuyuki Takeyasu, Takashi Kaneta
  • Journal Title: Analytical Sciences Volume: 印刷中
  • NAID: 130005696783
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Miniaturized chemiluminescence detection system for a microfluidic paper-based analytical device and its application to the determination of chromium (III) (2016)
  • Author(s): Waleed Alahmad, Kanchana Uraisin, Duangjai Nacapricha, Takashi Kaneta
  • Journal Title: Analytical Methods Volume: 8 Issue: 27 Pages: 5414-5420
  • DOI: 10.1039/c6ay00954a
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Chelate Titrations of Ca2+ and Mg2+ Using Microfluidic Paper-Based Analytical Devices (2016)
  • Author(s): Shingo Karita, Takashi Kaneta
  • Journal Title: Analytica Chimica Acta Volume: 924 Pages: 60-67
  • DOI: 10.1016/j.aca.2016.04.019
  • NAID: 120006484387
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Determination of Iron Ion in the Water of a Natural Hot Spring Using Microfluidic Paper-based Analytical Devices (2016)
  • Author(s): Kazuma Ogawa, Takashi Kaneta
  • Journal Title: Analytical Sciences Volume: 32 Issue: 1 Pages: 31-34
  • DOI: 10.2116/analsci.32.31
  • NAID: 130005118559
  • ISSN: 0910-6340, 1348-2246
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Direct determination of lignin peroxidase released from Phanerochaete chrysosporium by in-capillary enzyme assay using micellar electrokinetic chromatography (2016)
  • Author(s): Airi Harada, Keiko Sasaki, Takashi Kaneta
  • Journal Title: Journal of Chromatography A Volume: 1440 Pages: 145-149
  • DOI: 10.1016/j.chroma.2016.02.062
  • NAID: 120005725657
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Capillary Electrophoresis with Laser-induced Fluorescence Detection for Application in Intracellular Investigation of Anthracyclines and Multidrug Resistance Proteins (2015)
  • Author(s): Julius Mbuna, Takashi Kaneta
  • Journal Title: Analytical Sciences Volume: 31 Issue: 11 Pages: 1121-1128
  • DOI: 10.2116/analsci.31.1121
  • NAID: 130005108604
  • ISSN: 0910-6340, 1348-2246
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Acid-Base Titrations Using Microfluidic Paper-Based Analytical Devices (2014)
  • Author(s): Shingo Karita, Takashi Kaneta
  • Journal Title: Analytical Chemistry Volume: 86 Issue: 24 Pages: 12108-12114
  • DOI: 10.1021/ac5039384
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 光圧によるエクソソームの高効率捕集とがん診断法への応用 (2017)
  • Author(s): 金田 隆
  • Organizer: 「光圧によるナノ物質操作と秩序の創生」第1回公開シンポジウム
  • Place of Presentation: 千葉大学（千葉）
  • Year and Date: 2017-01-17
• [Presentation] Observation of cell blebs immobilized on a glass substrate by total internal reflection fluorescence microscopy (2016)
  • Author(s): Mika Isoyama, Takashi Kaneta
  • Organizer: Asianalysis XIII
  • Place of Presentation: Chaing Mai (Thailand)
  • Year and Date: 2016-12-08
  • Int'l Joint Research
• [Presentation] Collection of nano-vesicles using radiation pressure (2016)
  • Author(s): Mai Kuboi, Takashi Kaneta
  • Organizer: Asianalysis XIII
  • Place of Presentation: Chaing Mai (Thailand)
  • Year and Date: 2016-12-08
  • Int'l Joint Research
• [Presentation] Enzyme assay of manganese peroxidase by capillary electrophoresis (2016)
  • Author(s): Airi Harada, Sumire Kudo, Keiko Sasaki, Takashi Kaneta
  • Organizer: Asianalysis XIII
  • Place of Presentation: Chaing Mai (Thailand)
  • Year and Date: 2016-12-08
  • Int'l Joint Research
• [Presentation] Paper-based analytical devices for titrations in environmental chemistry and food chemistry (2016)
  • Author(s): Takashi Kaneta
  • Organizer: Asianalysis XIII
  • Place of Presentation: Chaing Mai (Thailand)
  • Year and Date: 2016-12-08
  • Int'l Joint Research / Invited
• [Presentation] キャピラリー電気泳動によるマンガンペルオキシダーゼの酵素アッセイ (2016)
  • Author(s): 金田 隆，工藤 すみれ，原田 愛梨，笹木 圭子
  • Organizer: 第36回キャピラリー電気泳動シンポジウム
  • Place of Presentation: 徳島大学（徳島）
  • Year and Date: 2016-11-09
• [Presentation] ペーパー分析デバイスで何ができるか (2016)
  • Author(s): 金田 隆
  • Organizer: 日本分析化学会第65年会
  • Place of Presentation: 北海道大学（札幌）
  • Year and Date: 2016-09-14
  • Invited
• [Presentation] ガラス基板上への細胞小胞固定の全反射蛍光顕微鏡観察 (2016)
  • Author(s): 礒山 美華，金田 隆
  • Organizer: 日本分析化学会第65年会
  • Place of Presentation: 北海道大学（札幌）
  • Year and Date: 2016-09-14
• [Presentation] ペキャピラリー電気泳動法によるマンガンペルオキシダーゼの活性測定法の開発 (2016)
  • Author(s): 工藤 すみれ，原田 愛梨，笹木 圭子，金田 隆
  • Organizer: 日本分析化学会第65年会
  • Place of Presentation: 北海道大学（札幌）
  • Year and Date: 2016-09-14
• [Presentation] レーザー光の輻射圧を利用する小胞の捕集 (2016)
  • Author(s): 久保井 麻衣， 金田 隆
  • Organizer: 日本分析化学会第65年会
  • Place of Presentation: 北海道大学（札幌）
  • Year and Date: 2016-09-14
• [Presentation] マイクロペーパー分析デバイス用小型化学発光検出器の開発 (2016)
  • Author(s): 金田隆，Waleed Alahmad
  • Organizer: 第76回分析化学討論会
  • Place of Presentation: 岐阜薬科大学（岐阜）
  • Year and Date: 2016-05-28
• [Presentation] Paper-based analytical devices for rapid titrations (2015)
  • Author(s): Takashi Kaneta
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015
  • Place of Presentation: Honolulu. USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research / Invited
• [Presentation] Determination of Fe(III) in hot spring water using microfluidic paper-based analytical devices (2015)
  • Author(s): Kazuma Ogawa, Takashi Kaneta
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015
  • Place of Presentation: Honolulu. USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research
• [Presentation] Chelate titrations using microfluidic paper-based analytical devices (2015)
  • Author(s): Shingo Karita, Takashi Kaneta
  • Organizer: The International Chemical Congress of Pacific Basin Societies 2015
  • Place of Presentation: Honolulu. USA
  • Year and Date: 2015-12-15
  • Int'l Joint Research
• [Presentation] マイクロ流体ペーパー分析デバイスによる天然水の簡易計測 (2015)
  • Author(s): 尾川 冬馬, 金田 隆
  • Organizer: 2015年日本化学会中国四国支部大会
  • Place of Presentation: 岡山大学
  • Year and Date: 2015-11-14
• [Presentation] キレート滴定用マイクロペーパー分析デバイス (µPAD) の開発 (2015)
  • Author(s): 苅田真吾，金田 隆
  • Organizer: 2015年日本化学会中国四国支部大会
  • Place of Presentation: 岡山大学
  • Year and Date: 2015-11-14
• [Presentation] キャピラリー電気泳動法によるリグニンペルオキシダーゼの活性測定法の開発 (2015)
  • Author(s): 原田愛梨，笹木圭子，金田隆
  • Organizer: 2015年日本化学会中国四国支部大会
  • Place of Presentation: 岡山大学
  • Year and Date: 2015-11-14
• [Presentation] ミセル動電クロマトグラフィーによるリグニンペルオキシダーゼの酵素アッセイ (2015)
  • Author(s): 原田 愛梨，笹木 圭子，金田 隆
  • Organizer: 第35回キャピラリー電気泳動シンポジウム
  • Place of Presentation: 岡山大学
  • Year and Date: 2015-11-04
• [Presentation] マイクロ流体ペーパー分析デバイス（µPADs）によるキレート滴定分析法の開発 (2015)
  • Author(s): 苅田 真吾，金田 隆
  • Organizer: 第35回キャピラリー電気泳動シンポジウム
  • Place of Presentation: 岡山大学
  • Year and Date: 2015-11-04
• [Presentation] キャピラリー電気泳動法によるリグニンペルオキシダーゼの活性評価 (2015)
  • Author(s): 原田 愛梨，笹木 圭子，金田 隆
  • Organizer: 日本分析化学会第64年会
  • Place of Presentation: 九州大学
  • Year and Date: 2015-09-09
• [Presentation] マイクロペーパー分析デバイスによるキレート滴定分析 (2015)
  • Author(s): 苅田 真吾，金田 隆
  • Organizer: 日本分析化学会第64年会
  • Place of Presentation: 九州大学
  • Year and Date: 2015-09-09
• [Presentation] Planarization of cell membrane on supported lipid bilayers (2015)
  • Author(s): 金田 隆，牧 朋美
  • Organizer: 日本分析化学会第64年会
  • Place of Presentation: 九州大学
  • Year and Date: 2015-09-09
  • Invited
• [Presentation] マイクロペーパー分析デバイスを用いたオンサイト化学計測 (2015)
  • Author(s): 金田 隆
  • Organizer: 日本分析化学会第64年会
  • Place of Presentation: 九州大学
  • Year and Date: 2015-09-09
  • Invited
• [Presentation] Rapid Acid-Base Titrations Using Microfluidic Paper-Based Analytical Devices (2015)
  • Author(s): Shingo Karita, Takashi Kaneta
  • Organizer: 2015 BK21Plus Symposium on HRD Center for Creative Convergence Chemical Science & The 2nd SKKU-OU Joint Symposium on Advanced Chemistry
  • Place of Presentation: Vivaldi Park & Ski Resort, Hongcheon. Korea
  • Year and Date: 2015-02-13
• [Presentation] 濃縮フィルターによる金属イオン検出用マイクロペーパー分析デバイスの高感度化 (2014)
  • Author(s): 尾川 冬馬，武安 伸幸，金田 隆
  • Organizer: 日本分析化学会第63年会
  • Place of Presentation: 広島大学
  • Year and Date: 2014-09-18
• [Presentation] 中和滴定用マイクロペーパー分析デバイスの開発 (2014)
  • Author(s): 苅田 真吾，武安 伸幸，金田 隆
  • Organizer: 日本分析化学会第63年会
  • Place of Presentation: 広島大学
  • Year and Date: 2014-09-18
• [Book] Capillary Electrophoresis of Proteins and Peptides, Methods and Protocols (2016)
  • Author(s): Takashi Kaneta（分筆）
  • Total Pages: 233
  • Publisher: Springer