研究者データベース
池袋 一典IKEBUKURO Kazunoriイケブクロ カズノリ
| ■所属部署名 | 工学研究院 生命機能科学部門 | |||
| ■職名 | 教授 | |||
Last Updated :2024/09/26
業績情報
氏名・連絡先
氏名
イケブクロ カズノリ, 池袋 一典, IKEBUKURO Kazunorieメールアドレス
ikebu
cc.tuat.ac.jp個人ホームページ
http://www.tuat.ac.jp/~tanpaku/
主たる所属・職名
工学研究院 生命機能科学部門, 教授
その他の所属
経歴
- -
東京大学先端科学技術研究センター 助手
自 1993年04月, 至 1996年03月 - -
東京大学先端科学技術研究センター 講師
自 1996年04月, 至 2001年03月 - -
東京農工大学工学部 助教授
自 2001年04月 - -
東京農工大学大学院工学研究院
自 2009年04月01日
学歴
教育・研究活動状況
- 私たちの身体の中では、遺伝情報を担うDNAと生理機能を担う蛋白質がお互いに作用して、生命現象を維持しています。私たちはDNAと蛋白質の相互作用を応用して、様々な病気の診断技術の開発を目的に研究を行っています。蛋白質を認識する核酸分子“アプタマー” を使って、癌やアルツハイマー病、感染症を診断する新規バイオセンサーの開発を進めています。また、核酸を認識する蛋白質“ジンクフィンガー” を使って、病原性微生物を検出するシステムの開発を進めています。
研究分野
研究キーワード
研究テーマ
- ＤＮＡ／ＳＮＰセンシングシステムの開発
自 1996 - 進化的手法を用いた新規DNA認識素子の開発
自 1997 - 新規進化的手法を用いた新規DNAアプタマーの開発
自 1998 - in silico panning 法によるペプチドリガンドの探索
自 200104 - アプタマーを用いた新規分子検出法の開発
自 2002
共同・受託研究希望テーマ
科学研究費助成事業
- 基盤研究(C)
疾患関連抗体に対する抗イディオタイプDNAアプタマーのファスト＆ファインデザイン
自 2022年, 至 2022年 - 挑戦的研究（萌芽）
細胞内の標的蛋白質を可視化するturn-on型蛍光RNAアプタマーの開発
自 2021年, 至 2022年 - 基盤研究(B)
抗体特異的な核酸アプタマーの創出とバイオ・プロセス・クリニカルアナリシスへの活用
自 2021年, 至 2021年 - 基盤研究(B)
抗体特異的な核酸アプタマーの創出とバイオ・プロセス・クリニカルアナリシスへの活用
自 2020年, 至 2020年 - 基盤研究(C)
分子内/分子間Gカルテット構造形成による免疫活性化CpG ODNの機能制御
自 2020年, 至 2020年 - 基盤研究(B)
光架橋性グアニン四重鎖リガンドを用いたメチローム構成蛋白質解析技術の開発
自 2019年, 至 2021年 - 挑戦的研究（萌芽）
気相中で標的分子を特異的に認識するDNAアプタマーの開発
自 2019年, 至 2020年 - 基盤研究(B)
抗体特異的な核酸アプタマーの創出とバイオ・プロセス・クリニカルアナリシスへの活用
自 2019年, 至 2019年 - 基盤研究(C)
分子内/分子間Gカルテット構造形成による免疫活性化CpG ODNの機能制御
自 2019年, 至 2019年 - 基盤研究(C)
分子内/分子間Gカルテット構造形成による免疫活性化CpG ODNの機能制御
自 2018年, 至 2018年 - 基盤研究(C)
眼内VEGF濃度の即時測定系開発と臨床応用?
自 2017年, 至 2017年 - 基盤研究（C）
眼内VEGF濃度の即時測定系開発と臨床応用
自 2016年, 至 2016年 - 基盤研究（A）
ナノニードルを用いた核輸送による高効率ゲノム編集
自 2016年, 至 2016年 - 挑戦的萌芽研究
細胞内での異常な核酸分子挙動の解析
自 2015年, 至 2015年 - 基盤研究（C）
眼内VEGF濃度の即時測定系開発と臨床応用
自 2015年, 至 2015年 - 基盤研究（A）
ナノニードルを用いた核輸送による高効率ゲノム編集
自 2015年, 至 2015年 - 挑戦的萌芽研究
細胞内での異常な核酸分子挙動の解析
自 2014年, 至 2014年 - 基盤研究(A)
ナノニードルを用いた核輸送による高効率ゲノム編集
自 2014年, 至 2014年 - 挑戦的萌芽研究
DNA折り紙を応用した新しい抗ウイルス薬の開発
自 2013年, 至 2013年 - 基盤研究(B)
アプタマーセットを用いた血液中極微量疾病マーカーの多数同時検出法の開発
自 2012年, 至 2014年 - 挑戦的萌芽研究
電気化学的グルコースセンサーとアプタマーのナノ構造変化を利用した検出法の開発
自 2012年, 至 2013年 - 挑戦的萌芽研究
Ｚｎフィンガー蛋白質を用いたメチル化頻度解析法の開発
自 2010年, 至 2011年 - 基盤研究(B)
ピロロキノリンキノンによるアミロイド蛋白質の選択的構造形成制御
自 2010年, 至 2010年 - 基盤研究(B)
ピロロキノリンキノンによるアミロイド蛋白質の選択的構造形成制御
自 2009年, 至 2009年 - 萌芽研究
磁場による誘導加熱を利用した金ナノ粒子修飾アプタマーの構造・機能制御
自 2008年, 至 2009年 - 基盤研究(B)
ピロロキノリンキノンによるアミロイド蛋白質の選択的構造形成制御
自 2008年, 至 2008年 - 基盤研究(C)一般
複数の疾病マーカー分子を特異的に認識するアプタマーの同時探索法の開発
自 2006年, 至 2007年 - 基盤研究(C)一般
複数の疾病マーカー分子を特異的に認識するアプタマーの同時探索法の開発
自 2006年, 至 2007年 - 若手研究(B)
コンピュータ内進化による疾病原因酵素阻害ＤＮＡアプタマ－の検索
自 2003年, 至 2004年 - 若手研究(B)
コンピューター内進化による疾病原因酵素阻害DNAアプタマーの探索
自 2003年, 至 2004年 - 基盤研究(C)一般
有害微生物のDNA検出のための新規ジンクフィンガー蛋白質の創出
自 2001年, 至 2002年 - その他
超機能バイオセンサーの開発
自 1998年, 至 1999年 - 基盤研究（C）
疾患関連抗体に対する抗イディオタイプDNAアプタマーのファスト＆ファインデザイン
自 2023年, 至 2023年 - 挑戦的研究（萌芽）
DNA中の疎水場を利用したナノプラスチック検出センサの開発
自 2023年, 至 2023年 - 基盤研究(B)
酵素・抗体・アプタマーの近接を利用した単一工程電気化学センサーの開発
自 2023年, 至 2025年
論文
- Structure of lactate oxidase from Enterococcus hirae revealed new aspects of active site loop function: Product-inhibition mechanism and oxygen gatekeeper
Hiraka, Kentaro; Yoshida, Hiromi; Tsugawa, Wakako; Asano, Ryutaro; La Belle, Jeffrey T.; Ikebukuro, Kazunori; Sode, Koji
PROTEIN SCIENCE
WILEY
l-Lactate oxidase (LOx) is a flavin mononucleotide (FMN)-dependent triose phosphate isomerase (TIM) barrel fold enzyme that catalyzes the oxidation of l-lactate using oxygen as a primary electron acceptor. Although reductive half-reaction mechanism of LOx has been studied by structure-based kinetic studies, oxidative half-reaction and substrate/product-inhibition mechanisms were yet to be elucidated. In this study, the structure and enzymatic properties of wild-type and mutant LOxs from Enterococcus hirae (EhLOx) were investigated. EhLOx structure showed the common TIM-barrel fold with flexible loop region. Noteworthy observations were that the EhLOx crystal structures prepared by co-crystallization with product, pyruvate, revealed the complex structures with d-lactate form ligand, which was covalently bonded with a Tyr211 side chain. This observation provided direct evidence to suggest the product-inhibition mode of EhLOx. Moreover, this structure also revealed a flip motion of Met207 side chain, which is located on the flexible loop region as well as Tyr211. Through a saturation mutagenesis study of Met207, one of the mutants Met207Leu showed the drastically decreased oxidase activity but maintained dye-mediated dehydrogenase activity. The structure analysis of EhLOx Met207Leu revealed the absence of flipping in the vicinity of FMN, unlike the wild-type Met207 side chain. Together with the simulation of the oxygen-accessible channel prediction, Met207 may play as an oxygen gatekeeper residue, which contributes oxygen uptake from external enzyme to FMN. Three clades of LOxs are proposed based on the difference of the Met207 position and they have different oxygen migration pathway from external enzyme to active center FMN.
2022年10月, 研究論文（学術雑誌）, 共同, 31, 10, 0961-8368, DOI(公開)(r-map) - Stabilization of VEGF i-motif structure by CpG methylation
Kimura, Kosuke; Oshikawa, Daiki; Ikebukuro, Kazunori; Yoshida, Wataru
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
ACADEMIC PRESS INC ELSEVIER SCIENCE
The intercalated motif (i-motif) is a non-canonical nucleic acid structure formed by intercalated hemiprotonated cytosine base pairs (C-C+) under acidic conditions. The i-motif structure formation is involved in biological processes such as transcription regulation. Therefore, the identification of factors controlling i-motif formation is important in elucidating the cellular functions it controls. We previously reported that the VEGF G-quadruplex structure is stabilized by CpG methylation. In this study, the effect of CpG methylation on the stability of the VEGF i-motif structure was investigated. The VEGF i-motifforming oligonucleotide contains four cytosines on CpG sites, and three of the four cytosines (C4, C15, and C20) are involved in C-C+ formation in the i-motif structure. Circular dichroism (CD) spectra analysis demonstrated that full CpG methylation increased the pH of mid transition (pHT) of the i-motif structure by 0.1, and the melting temperature (Tm) by 5.1 degrees C in 25 mM sodium cacodylate buffer at pH 5.0. Moreover, single methylation at C4, C15, and C20 increased Tm by 0.5, 1.7, and 2.0 degrees C in the buffer, respectively. These results demonstrated that CpG methylation stabilized the VEGF i-motif structure. (c) 2022 Elsevier Inc. All rights reserved.
2022年02月26日, 研究論文（学術雑誌）, 共同, 594, 0006-291X, DOI(公開)(r-map), 88, 92 - Development of a POCT type insulin sensor employing anti-insulin single chain variable fragment based on faradaic electrochemical impedance spectroscopy under single frequency measurement
Khanwalker, Mukund; Fujita, Rinko; Lee, Jinhee; Wilson, Ellie; Ito, Kohei; Asano, Ryutaro; Ikebukuro, Kazunori; LaBelle, Jeffrey; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
To improve glycemic control managed through insulin administration, recent studies have focused on developing hand-held point-of-care testing (POCT) electrochemical biosensors for insulin measurement. Amongst them, anti insulin IgG-based sensors show promise in detecting insulin with high specificity and sensitivity. However, fabrication of electrochemical sensors with IgG antibodies can prove challenging because of their larger molecular size. To overcome these limitations, this study focuses on utilizing the anti-insulin single chain variable fragment (scFv) as a biosensing molecule with single-frequency faradaic electrochemical impedance spectroscopy (EIS). By comparing two different immobilization methods, covalent conjugation via succinimidyl ester and non-covalent poly-histidine chelation, we demonstrated effective modification of the electrode surface with anti insulin scFv, while retaining its specific recognition toward insulin. Sensor performance was confirmed via the concentration-dependent faradaic electrochemical impedance change using potassium ferricyanide as a redox probe. The optimal frequency for measurement was determined to be the peak slope of the calculated impedance correlation with respect to frequency. Based on the identified optimized frequency, we performed single frequency measurement of insulin within a concentration range of 10 pM-100 nM. This study can aid in developing a future point-of-care sensor which rapidly and sensitively measures insulin across a dynamic range of physiological concentrations, with label-free detection.
2022年03月15日, 研究論文（学術雑誌）, 共同, 200, 0956-5663, DOI(公開)(r-map) - Transient potentiometry based D-serine sensor using engineered D-amino acid oxidase showing quasi-direct electron transfer property
Takamatsu, Shouhei; Lee, Inyoung; Lee, Jinhee; Asano, Ryutaro; Tsugawa, Wakako; Ikebukuro, Kazunori; Dick, Jeffrey E.; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
D-Serine biosensing has been extensively reported based on enzyme sensors using flavin adenine dinucleotide (FAD) -dependent n-amino acid oxidase (DAAOx), based on the monitoring of hydrogen peroxide generated by the enzymatic reaction, which is affected by dissolved oxygen concentration in the measurement environment in in vivo use. Here we report a novel sensing principle for o-serine, transient potentiometry based o-serine sensor using engineered DAAOx showing quasi-direct electron transfer (DET) property. DAAOx Gly52Val mutant, revealed to possess dye-mediated dehydmgenase activity using artificial synthetic electron acceptors, while its oxidase activity was negligible. The enzyme was immobilized on electrode and was modified with amine-reactive phenazine ethosulfate, resulted an enzyme electrode showing quasi-DET type response. Although OCP based monitoring took more than several minutes to obtain steady state OCP value, the time dependent OCP change monitoring, transient potentiometry, provided rapid and sensitive sensor signals. While dOCP/dt based monitoring was suitable for sensing with longer than 5 s time resolution with o-serine concentration range between 0.5 mM and 5 mM, dOCP/d root t based monitoring is suitable for o-serine monitoring with much shorter time resolution (less than 1 s) with high sensitivity with wider dynamic range (20 mu M-30 mM). The maximum dOCP/d root t was -39.2 +/- 2.0 mV/s(1/2), the K-m(app) was 1.9 mM, and the lower limit of detection was 20 mu M. In addition, D-serine monitoring was also possible in the artificial cerebrospinal fluid. The transient potentiometry based sensing reported in this study will be further utilized to realize miniaturized, continuous, real-time, in vivo sensor for o-serine monitoring.
2022年03月15日, 研究論文（学術雑誌）, 共同, 200, 0956-5663, DOI(公開)(r-map) - Development of a DNA aptamer that binds to the complementarity-determining region of therapeutic monoclonal antibody and affinity improvement induced by pH-change for sensitive detection
Saito, Taro; Shimizu, Yutaka; Tsukakoshi, Kaori; Abe, Koichi; Lee, Jinhee; Ueno, Kinuko; Asano, Ryutaro; Jones, Brian, V; Yamada, Tomohiro; Nakano, Tatsuki; Tong, Jiaxing; Hishiki, Asami; Hara, Kodai; Hashimoto, Hiroshi; Sode, Koji; Toyo'oka, Toshimasa; Todoroki, Kenichiro; Ikebukuro, Kazunori
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
Therapeutic monoclonal antibodies (mAbs) are successful biomedicines; however, evaluation of their pharmacokinetics and pharmacodynamics demands highly specific discrimination from human immunoglobulin G naturally present in the blood. Here, we developed a novel anti-idiotype aptamer (termed A14#1) with extraordinary specificity against the anti-vascular endothelial growth factor therapeutic mAb, bevacizumab. Structural analysis of the antibody-aptamer complex showed that several bases of A14#1 recognized only the complementarity determining region (CDR) of bevacizumab, thereby contributing to its extraordinary specificity. As the CDR of bevacizumab is predicted to be highly positively charged under mildly acidic conditions and that DNA is negatively charged, the affinity of A14#1 to bevacizumab markedly increased at pH 4.7 (K-D = 44 pM) than at pH 7.4 (K-D = 12 nM). A14#1-based electrochemical detection method capable of detecting 31 pM of bevacizumab at pH 4.7 was thus developed. A14#1 could be potentially useful for therapeutic drug measurement as a novel ligand of bevacizumab.
2022年05月01日, 研究論文（学術雑誌）, 共同, 203, 0956-5663, DOI(公開)(r-map) - The state of water molecules induces changes in the topologies and interactions of G-quadruplex DNA aptamers in hydrated ionic liquid
Fujita, Kyoko; Honda, Takuya; Tsukakoshi, Kaori; Ohno, Hiroyuki; Ikebukuro, Kazunori
JOURNAL OF MOLECULAR LIQUIDS
ELSEVIER
Herein, the effects of the bound state of water molecules were investigated on the topologies and inter-actions of G-quadruplex DNA aptamers by using hydrated ionic liquids (ILs). Since intracellular water molecules are generally expected to exist in the bound state and not as free molecules that exist in the typical in vitro media, we proposed hydrated ILs as potential candidates for controlling the state of water molecules. Hydrated ILs have been reported to dissolve proteins without compromising their higher-order structures. In this study, the structures and interactions of three G-quadruplex DNA apta-mers (one thrombin-binding aptamer and two vascular endothelial growth factor 165-binding aptamers), with their target molecules were analyzed with circular dichroism (CD) spectroscopy and enzyme-linked oligonucleotide assay by changing the water content of hydrated cholinium dihydrogen phosphate (Hy [ch][dhp]). Hy[ch][dhp] allowed the effective dissolution of G-quadruplex DNA aptamers while maintain-ing their structure and binding affinity to the target molecule. The water content of Hy[ch][dhp] induced changes in the CD spectra, suggesting changes in the topology of G-quadruplex structure. Increased struc-tural stability and binding properties indicated molecular recognition and smooth dehydration progress in Hy[ch][dhp] via regulation of the state of water molecules. (c) 2022 Elsevier B.V. All rights reserved.
2022年11月15日, 研究論文（学術雑誌）, 共同, 366, 0167-7322, DOI(公開)(r-map) - Effects of G-Quadruplex Ligands on the Topology, Stability, and Immunostimulatory Properties of G-Quadruplex-Based CpG Oligodeoxynucleotides
Tu, Anh Thi Tram; Hoshi, Kazuaki; Ma, Yue; Oyama, Taiji; Suzuki, Satoko; Tsukakoshi, Kaori; Nagasawa, Kazuo; Ikebukuro, Kazunori; Yamazaki, Tomohiko
ACS CHEMICAL BIOLOGY
AMER CHEMICAL SOC
We previously reported that the formation of guanine-quadruplex (G4) structures provides phosphodiester oligodeoxynucleotides containing unmethylated cytosine-phosphate-guanine (CpG ODNs) with higher nuclease resistance and cellular uptake, thereby increasing their immunostimulation efficiency through TLR9 activation. CpG ODNs forming G4 structures (G4 CpG ODNs) are thus potential vaccine adjuvants against infectious diseases. However, the G4 structure changes topology depending on the surrounding environment. Recently, G4 ligands, which are small molecules that bind to G4 ODNs with high affinity, were reported to improve the stability of G4. In this study, we propose to increase the stability and function of G4 CpG ODNs using G4 ligands. We show the effects of two G4 ligands, named L2H2-6OTD (L2H2) and L2G2-2M2EG-6OTD (L2G2), on the topology, stability, and immunostimulatory properties of a monomeric hybrid-type G4 CpG ODN containing CpG motifs in the central loop, named GD3. We found that L2H2 helps maintain the hybrid G4 topology of GD3, whereas L2G2 induces parallel G4 formation. Both G4 ligands increase the thermodynamic and nuclease stability of GD3. However, only GD3 associated with L2H2 binds efficiently to TLR9 and evokes a higher immune response from mouse macrophage-like RAW264 cells. GD3 associated with L2G2 does not bind efficiently to TLR9 and elicits lower cytokine production. Our results demonstrate that the potential to enhance immunostimulatory properties depends on the ability of G4 ligands to maintain and stabilize the hybrid G4 of GD3. We anticipate that our findings will facilitate the development of more effective G4 CpG ODN-based vaccine adjuvants against infectious diseases.
2022年07月15日, 研究論文（学術雑誌）, 共同, 17, 7, 1554-8929, DOI(公開)(r-map), 1703, 1713 - Light-induced production of isobutanol and 3-methyl-1-butanol by metabolically engineered cyanobacteria
Kobayashi, Shunichi; Atsumi, Shota; Ikebukuro, Kazunori; Sode, Koji; Asano, Ryutaro
MICROBIAL CELL FACTORIES
BMC
Background: Cyanobacteria are engineered via heterologous biosynthetic pathways to produce value-added chemicals via photosynthesis. Various chemicals have been successfully produced in engineered cyanobacteria. Chemical inducer-dependent promoters are used to induce the expression of target biosynthetic pathway genes. A chemical inducer is not ideal for large-scale reactions owing to its high cost; therefore, it is important to develop scaling-up methods to avoid their use. In this study, we designed a green light-inducible alcohol production system using the CcaS/CcaR green light gene expression system in the cyanobacterium Synechocystis sp. PCC 6803 (PCC 6803). Results: To establish the green light-inducible production of isobutanol and 3-methyl-1-butanol (3MB) in PCC 6803, keto-acid decarboxylase (kdc) and alcohol dehydrogenase (adh) were expressed under the control of the CcaS/CcaR system. Increases in the transcription level were induced by irradiation with red and green light without severe effects on host cell growth. We found that the production of isobutanol and 3MB from carbon dioxide (CO2) was induced under red and green light illumination and was substantially repressed under red light illumination alone. Finally, production titers of isobutanol and 3MB reached 238 mg L-1 and 75 mg L-1, respectively, in 5 days under red and green light illumination, and these values are comparable to those reported in previous studies using chemical inducers. Conclusion: A green light-induced alcohol production system was successfully integrated into cyanobacteria to produce value-added chemicals without using expensive chemical inducers.The green light-regulated production of isobutanol and 3MB from CO2 is eco-friendly and cost-effective. This study demonstrates that light regulation is a potential tool for producing chemicals and increases the feasibility of cyanobacterial bioprocesses.
2022年01月06日, 研究論文（学術雑誌）, 共同, 21, 1, DOI(公開)(r-map) - A Thiol-reactive Phenazine Ethosulfate - A Novel Redox Mediator for Quasi-direct Electron-transfer-type Sensors
Fitriana, Maya; Hiraka, Kentaro; Ikebukuro, Kazunori; Sode, Koji; Tsugawa, Wakako
SENSORS AND MATERIALS
MYU, SCIENTIFIC PUBLISHING DIVISION
A novel redox mediator, thiol-reactive phenazine ethosulfate (trPES), was used to modify an enzyme for the first time for biosensor development. Aerococcus viridans-lactate oxidase (LOx), widely used to study lactate biosensors, was modified with a single trPES molecule. A cysteine mutation was introduced into the vicinity of the LOx cofactor to enable the modification by trPES. LOx cysteine mutants were then successfully modified using trPES, thus acquiring quasi-direct electron transfer ability. An electrode immobilized with the trPES-LOx A96L/S210C mutant showed the highest amperometric response currents among the modified LOx cysteine mutants, indicating efficient electron transfer. The position around residues S210 to N212 on the LOx surface (distance <14 angstrom from FMN N5) is important for the mediator to access the reduced flavin. Then, the performances of the lactate sensor were improved by utilizing LOx A96L/S210C modified with trPES and arPES, another redox mediator used to modify a lysine residue. The lactate sensor has a detection range of up to 1 mM, a sensitivity of 6.62 mu A/mM.cm(2), and a limit of detection of 9.9 mu M. Furthermore, Aspergillus flavus-derived FAD glucose dehydrogenase was successfully modified with trPES and the response currents were obtained, showing the versatility of trPES for modifying other oxidoreductases.
2022年, 研究論文（学術雑誌）, 共同, 34, 6, 0914-4935, DOI(公開)(r-map), 2105, 2124 - CpG Methylation Altered the Stability and Structure of the i-Motifs Located in the CpG Islands
Oshikawa, Daiki; Inaba, Shintaro; Kitagawa, Yudai; Tsukakoshi, Kaori; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Cytosine methylation within the 5 '-C-phosphate-G-3 ' sequence of nucleotides (called CpG methylation) is a well-known epigenetic modification of genomic DNA that plays an important role in gene expression and development. CpG methylation is likely to be altered in the CpG islands. CpG islands are rich in cytosine, forming a structure called the i-motif via cytosine-cytosine hydrogen bonding. However, little is known about the effect of CpG methylation on the i-motif. In this study, The CpG methylation-induced structural changes on the i-motif was examined by thermal stability, circular dichroism (CD) spectroscopy, and native-polyacrylamide gel electrophoresis (Native-PAGE) evaluation of five i-motif-forming DNAs from four cancer-related genes (VEGF, C-KIT, BCL2, and HRAS). This research shows that CpG methylation increased the transitional pH of several i-motif-forming DNAs and their thermal stability. When examining the effect of CpG methylation on the i-motif in the presence of opposite G4-forming DNAs, CpG methylation influenced the proportion of G4 and i-motif formation. This study showed that CpG methylation altered the stability and structure of the i-motif in CpG islands.
2022年06月, 研究論文（学術雑誌）, 共同, 23, 12, DOI(公開)(r-map) - Development of Alkaline Phosphatase-Fused Mouse Prion Protein and Its Application in Toxic A beta Oligomer Detection
Tsukakoshi, Kaori; Kubo, Rikako; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Amyloid beta (A beta) oligomers play a key role in the progression of Alzheimer's disease (AD). Multiple forms of A beta assemblies have been identified by in vitro and in vivo analyses; however, it is uncertain which oligomer is highly neurotoxic. Thus, understanding the pathogenesis of AD by detecting toxic A beta oligomers is crucial. In this study, we report a fusion protein of cellular prion protein (PrPc) and alkaline phosphatase (ALP) from Escherichia coli as a sensing element for toxic A beta oligomers. Since the N-terminus domain of PrPc (residue 23-111) derived from mice is known to bind to toxic A beta oligomers in vitro, we genetically fused PrPc23-111 to ALP. The developed fusion protein, PrP-ALP, retained both the binding ability of PrPc and enzymatic activity of ALP. We showed that PrP-ALP strongly bound to high molecular weight (HMW) oligomers but showed little or no affinity toward monomers. The observation that PrP-ALP neutralized the toxic effect of A beta oligomers indicated an interaction between PrP-ALP and toxic HMW oligomers. Based on ALP activity, we succeeded in detecting A beta oligomers. PrP-ALP may serve as a powerful tool for detecting toxic A beta oligomers that may be related to AD progression.
2022年12月, 研究論文（学術雑誌）, 共同, 23, 23, DOI(公開)(r-map) - An Amine-Reactive Phenazine Ethosulfate (arPES)-A Novel Redox Probe for Electrochemical Aptamer-Based Sensor
Nagata, Madoka; Lee, Jinhee; Henley, Stephen; Ikebukuro, Kazunori; Sode, Koji
SENSORS
MDPI
Electrochemical aptamer-based biosensors (E-ABs) are attractive candidates for use in biomarker detection systems due to their sensitivity, rapid response, and design flexibility. There are only several redox probes that were employed previously for this application, and a combination of redox probes affords some advantages in target detection. Thus, it would be advantageous to study new redox probes in an E-AB system. In this study, we report the use of amine-reactive phenazine ethosulfate (arPES) for E-AB through its conjugation to the terminus of thrombin-binding aptamer. The constructed E-AB can detect thrombin by square-wave voltammetry (SWV), showing peak current at -0.15 V vs. Ag/AgCl at pH 7, which differs from redox probes used previously for E-ABs. We also compared the characteristics of PES as a redox probe for E-AB to methylene blue (MB), which is widely used. arPES showed stable signal at physiological pH. Moreover, the pH profile of arPES modified thrombin-binding aptamer revealed the potential application of arPES for a simultaneous multianalyte detection system. This could be achieved using different aptamers with several redox probes in tandem that harbor various electrochemical peak potentials. Our findings present a great opportunity to improve the current standard of biological fluid monitoring using E-AB.
2022年03月, 研究論文（学術雑誌）, 共同, 22, 5, DOI(公開)(r-map) - A Green Light-Regulated T7 RNA Polymerase Gene Expression System for Cyanobacteria
Shono, Chika; Ariyanti, Dwi; Abe, Koichi; Sakai, Yuta; Sakamoto, Ippei; Tsukakoshi, Kaori; Sode, Koji; Ikebukuro, Kazunori
MARINE BIOTECHNOLOGY
SPRINGER
In this study, we developed a green light-regulated T7 RNA polymerase expression system (T7 RNAP system), to provide a novel and versatile high-expression system for cyanobacteria without using any chemical inducer, realizing high expression levels comparable with previously reported for recombinant gene expression in cyanobacteria. The T7 RNAP system was constructed and introduced intoSynechocystissp. PCC6803. T7 RNAP was inserted downstream of thecpcG2promoter, which is recognized and activated by the CcaS/CcaR two-component green-light-sensing system, to compose a vector plasmid, pKT-CS01, to achieve the induction of T7 RNAP expression only under green light illumination, with repression under red light illumination. The reporter gene, superfolder green fluorescent protein (sfGFP), was inserted downstream of theT7promoter. Transcriptional analyses revealed that T7 RNAP was induced under green light but repressed under red light. Expression of thesfGFP protein derived from pKT-CS01 was observed under green light illumination and was approximately 10-fold higher than that in the control transformant, which expressedsfGFP directly under thecpcG2promoter, which is directly regulated by CcaS/CcaR, under green light illumination. Comparison with the strong promoter expression systems P(cpc560)and P(trc Delta lacO)revealed that the expression ofsfGFP by the T7 RNAP system was comparable with the levels obtained with strong promoters. These results demonstrated that the green light-regulated T7 RNAP gene expression system will be a versatile tool for future technological platform to regulate gene expression in cyanobacterial bioprocesses.
2021年02月, 研究論文（学術雑誌）, 共同, 23, 1, 1436-2228, DOI(公開)(r-map), 31, 38 - Rapid and homogeneous electrochemical detection by fabricating a high affinity bispecific antibody-enzyme complex using two Catcher/Tag systems
Kimura, Hayato; Miura, Daimei; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji; Asano, Ryutaro
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
Antibody-enzyme complexes (AECs) with binding ability to specific targets and catalytic activities to gain signals are known to be ideal sensing elements; however, AEC-based universal sensors applicable to point-of-care testing (POCT) have not yet been developed. Here, we achieved rapid and homogeneous electrochemical detection by fabricating a high-affinity bispecific AEC (bsAEC) using two Catcher/Tag systems. Recently, we reported a convenient and universal method to fabricate AECs using the SpyCatcher/SpyTag system. The resultant anti-epidermal growth factor receptor (anti-EGFR) AEC worked efficiently as a sensing element; however, the sensitivities did not meet the clinically required detection range of the soluble ectodomain of EGFR (sEGFR). To induce high affinity even to monomeric targets like sEGFR, we designed a convenient fabrication method for bsAEC using two Catcher/Tag systems, which did not express cross-reactivity. The anti-EGFR bsAEC was successfully prepared by constructing glucose dehydrogenase with two different catcher domains at the N- and C-terminus and by combining two corresponding Tag-fused anti-EGFR single-chain Fvs (scFvs), which recognize different epitopes on sEGFR. As expected, bsAEC showed a higher affinity than that of bivalent AEC with two identical anti-EGFR scFvs at low concentrations of sEGFR, and met the clinically required detection range of sEGFR. Further, by combining magnet beads, we established a rapid and wash-free homogeneous electrochemical detection method. This study offers new insights into the fabrication of universal POCT devices.
2021年03月01日, 研究論文（学術雑誌）, 共同, 175, 0956-5663, DOI(公開)(r-map) - Strategic design and improvement of the internal electron transfer of heme b domain-fused glucose dehydrogenase for use in direct electron transfer-type glucose sensors
Ito, Kohei; Okuda-Shimazaki, Junko; Kojima, Katsuhiro; Mori, Kazushige; Tsugawa, Wakako; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
A fusion enzyme composed of an Aspergillus flavus-derived flavin adenine dinucleotide glucose dehydrogenase (AfGDH) and an electron transfer domain of Phanerochaete chrysosporium-derived cellobiose dehydrogenase (Pcyb) was previously reported to show the direct electron transfer (DET) ability to an electrode. However, its slow intramolecular electron transfer (IET) rate from the FAD to the heme, limited the sensor signals. In this study, fusion FADGDH (Pcyb-AfGDH) enzymes were strategically redesigned by performing docking simulation, following surface-electrostatic potential estimation in the predicted area. Based on these predictions, we selected the amino acid substitution on Glu324, or on Asn408 to Lys to increase the positive charge at the rim of the interdomain region. Pcyb-AfGDH mutants were recombinantly produced using Pichia pastoris as the host microorganism, and their IET was evaluated. Spectroscopic observations showed that the Glu324Lys (E324K) and Asn408Lys (N408K) Pcyb-AfGDH mutants showed approximately 1.70and 9.0-fold faster IET than that of wildtype Pcyb-AfGDH, respectively. Electrochemical evaluation revealed that the mutant Pcyb-AfGDHimmobilized electrodes showed higher DET current values than that of the wildtype Pcyb-AfGDH-immobilized electrodes at pH 6.5, which was approximately 9-fold higher in the E324K mutant and 15-fold higher in the N408K mutant, than in the wildtype. Glucose enzyme sensors employing N408K mutant was able to measure glucose concentration under physiological condition using artificial interstitial fluid at pH 7.4, whereas the one with wildtype Pcyb-AfGDH was not. These results indicated that the sensor employed the redesigned mutant Pcyb-AfGDH can be used for future continuous glucose monitoring system based on direct electron transfer principle. (247 words).
2021年03月15日, 研究論文（学術雑誌）, 共同, 176, 0956-5663, DOI(公開)(r-map) - Rational design of direct electron transfer type L-lactate dehydrogenase for the development of multiplexed biosensor
Hiraka, Kentaro; Tsugawa, Wakako; Asano, Ryutaro; Yokus, Murat A.; Ikebukuro, Kazunori; Daniele, Michael A.; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
The development of wearable multiplexed biosensors has been focused on systems to measure sweat L-lactate and other metabolites, where the employment of the direct electron transfer (DET) principle is expected. In this paper, a fusion enzyme between an engineered L-lactate oxidase derived from Aerococcus viridans, AvLOx A96L/N212K mutant, which is minimized its oxidase activity and b-type cytochrome protein was constructed to realize multiplexed DET-type lactate and glucose sensors. The sensor with a fusion enzyme showed DET to a gold electrode, with a limited operational range less than 0.5 mM. A mutation was introduced into the fusion enzyme to increase K-m value and eliminate its substrate inhibition to construct b2LOxS. Together with the employment of an outer membrane, the detection range of the sensor with b2LOxS was expanded up to 10 mM. A simultaneous lactate and glucose monitoring system was constructed using a flexible thin-film multiplexed electrodes with b2LOxS and a DET-type glucose dehydrogenase, and evaluated their performance in the artificial sweat. The sensors achieved simultaneous detection of lactate and glucose without cross-talking error, with the detected linear ranges of 0.5-20 mM for lactate and 0.1-5 mM for glucose, sensitivities of 4.1 nA/mM.mm(2) for lactate and 56 nA/mM.mm(2) for glucose, and limit of detections of 0.41 mM for lactate and 0.057 mM for glucose. The impact of the presence of electrochemical interferants (ascorbic acid, acetaminophen and uric acid), was revealed to be negligible. This is the first report of the DET-type enzyme based lactate and glucose dual sensing systems.
2021年03月15日, 研究論文（学術雑誌）, 共同, 176, 0956-5663, DOI(公開)(r-map) - Development of glycated peptide enzyme sensor based flow injection analysis system for haemoglobin A1c monitoring using quasi-direct electron transfer type engineered fructosyl peptide oxidase
Hatada, Mika; Saito, Satomi; Yonehara, Satoshi; Tsugawa, Wakako; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
Haemoglobin A1e (hemoglobin A1e, HbA1c) is an important long-term glycemic control marker for diabetes. The aim of this study was to develop an enzyme flow injection analysis (FIA) system using engineered fructosyl peptide oxidase (FPOx) based on 2.5th generation principle for an HbA1c automated analytical system. FPOx from Phaeosphaeria nodorum (PnFPOx) was engineered by introducing a Lys residue at the R414 position, to be modified with amine reactive phenazine ethosulfate (arPES) in proximity of FAD. The engineered PnFPOx mutant with minimized oxidase activity, N56A/R414K, showed quasi-direct electron transfer (quasi-DET) ability after PES-modification. The FIA system was constructed by employing a PES-modified PnFPOx N56A/R414K and operated at 0 V against Ag/AgCl. The system showed reproducible responses with a linear range of 20-500 mu M for both fructosyl valine (FV) and fructosyl valylhistidine (FVH), with sensitivities of 0.49 nA mu AO and 0.13 nA mu M-1, and the detection limits of 1.3 mu M and 2.0 mu M for FV and FVH, respectively. These results indicate that the enzyme electrochemical FIA system covers the clinical range of HbA1c detection for more 200 consecutive measurements. Protease digested three different levels of HbA1c samples including healthy and diabetic range subjects were also measured with the FIA system. Thus, it will be possible to develop an integrated system consisting of sample pretreatment and sample electrochemical measurement based on an FIA system possessing quasi-DET type PnFPOx.
2021年04月01日, 研究論文（学術雑誌）, 共同, 177, 0956-5663, DOI(公開)(r-map) - G-quadruplex: Flexible conformational changes by cations, pH, crowding and its applications to biosensing
Nishio, Maui; Tsukakoshi, Kaori; Ikebukuro, Kazunori
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
G-quadruplex (G4) is a non-canonical structure that is formed in G-rich sequences of nucleic acids. G4s play important roles in vivo, such as telomere maintenance, transcription, and DNA replication. There are three typical topologies of G4: parallel, anti-parallel, and hybrid. In general, metal cations, such as potassium and sodium, stabilize G4s through coordination in the G-quartet. While G4s have some functions in vivo, there are many reports of developed applications that use G4s. As various conformations of G4s could form from one sequence depending on varying conditions, many researchers have developed G4-based sensors. Furthermore, G4 is a great scaffold of aptamers since many aptamers folded into G4s have also been reported. However, there are some challenges about its practical use due to the difference between practical sample conditions and experimental ones. G4 conformations are dramatically altered by the surrounding conditions, such as metal cations, pH, and crowding. Many studies have been conducted to characterize G4 conformations under various conditions, not only to use G4s in practical applications but also to reveal its function in vivo. In this review, we summarize recent studies that have investigated the effects of surrounding conditions (e.g., metal cations, pH, and crowding) on G4 conformations and the application of G4s mainly in biosensor fields, and in others.
2021年04月15日, 研究論文（学術雑誌）, 共同, 178, 0956-5663, DOI(公開)(r-map) - Continuous electrochemical monitoring of L-glutamine using redox-probe-modified L-glutamine-binding protein based on intermittent pulse amperometry
Takamatsu, Shouhei; Lee, Jinhee; Asano, Ryutaro; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji
SENSORS AND ACTUATORS B-CHEMICAL
ELSEVIER SCIENCE SA
The development of a continuous electrochemical monitoring system using a periplasmic binding protein (PBP), which changes its conformational dynamics upon ligand binding, is promising for in situ, real-time, continuous measurement technologies for use in effective biological production processes. This study focuses on a continuous L-glutamine biosensor based on Escherichia coli-derived L-glutamine-binding protein (QBP), a PBP that can speficically bind to L-glutamine. In this sensor, QBP was modified with a redox probe, amine-reactive phenazine ethosulfate (PES), and the conformational change in QBP through the recognition of L-glutamine was monitored electrochemically by intermittent pulse amperometry (IPA) based on the change in the access of QBP-modified PES to the electrode. This sensor exhibited a change in the current against logarithmic L-glutamine concentrations between 0.2-50 mu M. Continuous monitoring of L-glutamine, based on IPA measurements, revealed that continuous changes in L-glutamine concentration were observed for both increasing and decreasing concentrations. During continuous monitoring, L-glutamine concentration was monitored between 50 and 500 mu M, with a limit of detection of 50 mu M. This result is expected to address the future development of an electrochemical biosensor for in situ, real-time, continuous monitoring of various metabolites based on PBPs.
2021年11月01日, 研究論文（学術雑誌）, 共同, 346, DOI(公開)(r-map) - Rapid, convenient, and highly sensitive detection of human hemoglobin in serum using a high-affinity bivalent antibody-enzyme complex
Miura, Daimei; Kimura, Hayato; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji; Asano, Ryutaro
TALANTA
ELSEVIER
Human hemoglobin (Hb) is a biomarker of several diseases, and monitoring of Hb levels is required during emergent surgery. However, rapid and sensitive Hb detection methods are yet to be developed. The present study established a rapid, convenient, and highly sensitive detection method for Hb in human serum using a bivalent antibody-enzyme complex (AEC). AECs are promising sensing elements because of their ability to bind specific targets and their catalytic activity that produce signals. We recently reported a convenient and universal method to fabricate bivalent AECs with two antibody fragments, using the SpyCatcher/SpyTag system. The present study applied a bivalent AEC for highly sensitive and quantitative detection of human Hb. The bivalent anti-Hb AEC was successfully prepared by incubating both N- and C-terminus SpyCatcher-fused glucose dehydrogenase and SpyTag-fused anti-Hb single-chain variable fragments at 4 degrees C. As expected, the bivalent AEC for Hb with a multimeric structure showed higher affinity than the monovalent AEC, by means of avidity effects, unlike that for soluble epidermal growth factor receptor with a monomeric structure; this contributed to a great improvement in sensitivity. Finally, we established a rapid and wash-free homogeneous electrochemical detection system for Hb by integrating magnetic beads. The linear range of the system completely covered the clinically required Hb levels, even in human serum. This technology provides an ideal point-of-care test for Hb and other multimeric biomarkers.
2021年11月01日, 研究論文（学術雑誌）, 共同, 234, 0039-9140, DOI(公開)(r-map) - A self-powered glucose sensor based on BioCapacitor principle with micro-sized enzyme anode employing direct electron transfer type FADGDH
Lee, Inyoung; Okuda-Shimazaki, Junko; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji
JOURNAL OF PHYSICS-ENERGY
IOP PUBLISHING LTD
Diabetes mellitus is a disorder in which the body does not produce enough or respond normally to insulin; consequently, blood glucose levels increase to become abnormally high. Accordingly, the primary treatment of diabetes is to control glycemic levels continuously. To continuously control glycemic levels, several medical devices have been developed to monitor blood glucose levels, represented by sensors and monitors for the self-monitoring of blood glucose. The ultimate goal for those engaged in research to develop medical devices is to develop implantable biodevices, namely self-powered autonomously operated artificial pancreas systems. One of the most challenging issues in realizing an implantable artificial pancreas is the long-term continuous supply of electricity, which is currently dependent on rechargeable batteries, requiring periodical replacement. In this work, we report the development of a direct electron transfer type enzyme-based miniaturized self-powered glucose sensor based on the BioCapacitor principle with a micro-sized enzyme anode area (0.15 mm x 0.75 mm), which has only 0.1 mm(2) of electrode surface. As a result, a BioCapacitor utilizing a biofuel cell with a micro-sized enzyme anode was operated by self-power. In addition, the glucose concentration was detected within the range from 13 mM to 100 mM based on the frequency of charge/discharge cycles of the BioCapacitor. Although further improvement of the current density of the micro-sized anode is necessary to monitor a glucose concentration range lower than 13 mM, this self-powered glucose sensor with a micro-sized electrode based on the BioCapacitor principle was operated continuously for 6.6 h at 37 degrees C in 100 mM potassium phosphate buffer (pH 7.0). Our success indicates the potential to realize self-powered, autonomous, and implantable sensing modules for bio devices such as glucose-sensing systems for an artificial pancreas.
2021年07月, 研究論文（学術雑誌）, 共同, 3, 3, 2515-7655, DOI(公開)(r-map) - G-quadruplex-forming aptamer enhances the peroxidase activity of myoglobin against luminol
Tsukakoshi, Kaori; Yamagishi, Yasuko; Kanazashi, Mana; Nakama, Kenta; Oshikawa, Daiki; Savory, Nasa; Matsugami, Akimasa; Hayashi, Fumiaki; Lee, Jinhee; Saito, Taiki; Sode, Koji; Khunathai, Kanjana; Kuno, Hitoshi; Ikebukuro, Kazunori
NUCLEIC ACIDS RESEARCH
OXFORD UNIV PRESS
Aptamers can control the biological functions of enzymes, thereby facilitating the development of novel biosensors. While aptamers that inhibit catalytic reactions of enzymes were found and used as signal transducers to sense target molecules in biosensors, no aptamers that amplify enzymatic activity have been identified. In this study, we report G-quadruplex (G4)-forming DNA aptamers that upregulate the peroxidase activity in myoglobin specifically for luminol. Using in vitro selection, one G4-forming aptamer that enhanced chemiluminescence from luminol by myoglobin's peroxidase activity was discovered. Through our strategy-in silico maturation, which is a genetic algorithm-aided sequence manipulation method, the enhancing activity of the aptamer was improved by introducing mutations to the aptamer sequences. The best aptamer conserved the parallel G4 property with over 300-times higher luminol chemiluminescence from peroxidase activity more than myoglobin alone at an optimal pH of 5.0. Furthermore, using hemin and hemin-binding aptamers, we demonstrated that the binding property of the G4 aptamers to heme in myoglobin might be necessary to exert the enhancing effect. Structure determination for one of the aptamers revealed a parallel-type G4 structure with propeller-like loops, which might be useful for a rational design of aptasensors utilizing the G4 aptamer-myoglobin pair.
2021年06月21日, 研究論文（学術雑誌）, 共同, 49, 11, 0305-1048, DOI(公開)(r-map), 6069, 6081 - Artificial complementary chromatic acclimation gene expression system in Escherichia coli
Ariyanti, Dwi; Ikebukuro, Kazunori; Sode, Koji
MICROBIAL CELL FACTORIES
BMC
Background The development of multiple gene expression systems, especially those based on the physical signals, such as multiple color light irradiations, is challenging. Complementary chromatic acclimation (CCA), a photoreversible process that facilitates the control of cellular expression using light of different wavelengths in cyanobacteria, is one example. In this study, an artificial CCA systems, inspired by type III CCA light-regulated gene expression, was designed by employing a single photosensor system, the CcaS/CcaR green light gene expression system derived from Synechocystis sp. PCC6803, combined with G-box (the regulator recognized by activated CcaR), the cognate cpcG2 promoter, and the constitutively transcribed promoter, the P-trc Delta LacO promoter. Results One G-box was inserted upstream of the cpcG2 promoter and a reporter gene, the rfp gene (green light-induced gene expression), and the other G-box was inserted between the P-trc Delta LacO promoter and a reporter gene, the bfp gene (red light-induced gene expression). The Escherichia coli transformants with plasmid-encoded genes were evaluated at the transcriptional and translational levels under red or green light illumination. Under green light illumination, the transcription and translation of the rfp gene were observed, whereas the expression of the bfp gene was repressed. Under red light illumination, the transcription and translation of the bfp gene were observed, whereas the expression of the rfp gene was repressed. During the red and green light exposure cycles at every 6 h, BFP expression increased under red light exposure while RFP expression was repressed, and RFP expression increased under green light exposure while BFP expression was repressed. Conclusion An artificial CCA system was developed to realize a multiple gene expression system, which was regulated by two colors, red and green lights, using a single photosensor system, the CcaS/CcaR system derived from Synechocystis sp. PCC6803, in E. coli. The artificial CCA system functioned repeatedly during red and green light exposure cycles. These results demonstrate the potential application of this CCA gene expression system for the production of multiple metabolites in a variety of microorganisms, such as cyanobacteria.
2021年07月05日, 研究論文（学術雑誌）, 共同, 20, 1, DOI(公開)(r-map) - Enhancement of the Immunostimulatory Effect of Phosphodiester CpG Oligodeoxynucleotides by an Antiparallel Guanine-Quadruplex Structural Scaffold
Safitri, Fika Ayu; Tu, Anh Thi Tram; Hoshi, Kazuaki; Shobo, Miwako; Zhao, Dandan; Witarto, Arief Budi; Sumarsono, Sony Heru; Giri-Rachman, Ernawati Arifin; Tsukakoshi, Kaori; Ikebukuro, Kazunori; Yamazaki, Tomohiko
BIOMOLECULES
MDPI
Guanine-quadruplex-based CpG oligodeoxynucleotides (G4 CpG ODNs) have been developed as potent immunostimulatory agents with reduced sensitivity to nucleases. We designed new monomeric G4 ODNs with an antiparallel topology using antiparallel type duplex/G4 ODNs as robust scaffolds, and we characterized their topology and effects on cytokine secretion. Based on circular dichroism analysis and quantification of mRNA levels of immunostimulatory cytokines, it was found that monomeric antiparallel G4 CpG ODNs containing two CpG motifs in the first functional loop, named G2.0.0, could maintain antiparallel topology and generate a high level of immunostimulatory cytokines in RAW264 mouse macrophage-like cell lines. We also found that the flanking sequence in the CpG motif altered the immunostimulatory effects. Gc2c.0.0 and Ga2c.0.0 are monomeric antiparallel G4 CpG ODNs with one cytosine in the 3 & PRIME; terminal and one cytosine/adenine in the 5 & PRIME; terminal of CpG motifs that maintained the same resistance to degradation in serum as G2.0.0 and improved interleukin-6 production in RAW264 and bone marrow-derived macrophages. The immunostimulatory activity of antiparallel G4 CpG ODNs is superior to that of linear natural CpG ODNs. These results provide insights for the rational design of highly potent CpG ODNs using antiparallel G4 as a robust scaffold.
2021年11月, 研究論文（学術雑誌）, 共同, 11, 11, DOI(公開)(r-map) - Cytotoxic A beta Protofilaments Are Generated in the Process of A beta Fibril Disaggregation
Kaku, Toshisuke; Tsukakoshi, Kaori; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Significant research on Alzheimer's disease (AD) has demonstrated that amyloid beta (A beta) oligomers are toxic molecules against neural cells. Thus, determining the generation mechanism of toxic A beta oligomers is crucial for understanding AD pathogenesis. A beta fibrils were reported to be disaggregated by treatment with small compounds, such as epigallocatechin gallate (EGCG) and dopamine (DA), and a loss of fibril shape and decrease in cytotoxicity were observed. However, the characteristics of intermediate products during the fibril disaggregation process are poorly understood. In this study, we found that cytotoxic A beta aggregates are generated during a moderate disaggregation process of A beta fibrils. A cytotoxicity assay revealed that A beta fibrils incubated with a low concentration of EGCG and DA showed higher cytotoxicity than A beta fibrils alone. Atomic force microscopy imaging and circular dichroism spectrometry showed that short and narrow protofilaments, which were highly stable in the beta-sheet structure, were abundant in these moderately disaggregated samples. These results indicate that toxic A beta protofilaments are generated during disaggregation from amyloid fibrils, suggesting that disaggregation of A beta fibrils by small compounds may be one of the possible mechanisms for the generation of toxic A beta aggregates in the brain.
2021年12月, 研究論文（学術雑誌）, 共同, 22, 23, DOI(公開)(r-map) - Detection of CpG Methylation in G-Quadruplex Forming Sequences Using G-Quadruplex Ligands
Hasegawa, Hijiri; Sasaki, Ikkei; Tsukakoshi, Kaori; Ma, Yue; Nagasawa, Kazuo; Numata, Shusuke; Inoue, Yuuki; Kim, Yeji; Ikebukuro, Kazunori
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Genomic DNA methylation is involved in many diseases and is expected to be a specific biomarker for even the pre-symptomatic diagnosis of many diseases. Thus, a rapid and inexpensive detection method is required for disease diagnosis. We have previously reported that cytosine methylation in G-quadruplex (G4)-forming oligonucleotides develops different G4 topologies. In this study, we developed a method for detecting CpG methylation in G4-forming oligonucleotides based on the structural differences between methylated and unmethylated G4 DNAs. The differences in G4 topologies due to CpG methylation can be discriminated by G4 ligands. We performed a binding assay between methylated or unmethylated G4 DNAs and G4 ligands. The binding abilities of fluorescent G4 ligands to BCL-2, HRAS1, HRAS2, VEGF G4-forming sequences were examined by fluorescence-based microtiter plate assay. The differences in fluorescence intensities between methylated and unmethylated G4 DNAs were statistically significant. In addition to fluorescence detection, the binding of G4 ligand to DNA was detected by chemiluminescence. A significant difference was also detected in chemiluminescence intensity between methylated and unmethylated DNA. This is the first study on the detection of CpG methylation in G4 structures, focusing on structural changes using G4 ligands.
2021年12月, 研究論文（学術雑誌）, 共同, 22, 23, DOI(公開)(r-map) - Monomeric G-Quadruplex-Based CpG Oligodeoxynucleotides as Potent Toll-Like Receptor 9 Agonists
Tu, Anh Thi Tram; Hoshi, Kazuaki; Ikebukuro, Kazunori; Hanagata, Nobutaka; Yamazaki, Tomohiko
BIOMACROMOLECULES
AMER CHEMICAL SOC
Synthetic oligodeoxynucleotides (ODNs) containing unmethylated cytosine-phosphate-guanine (CpG) motifs trigger the immune response by stimulating endosomal Toll-like receptor (TLR) 9. Natural linear ODNs are susceptible to nuclease degradation, thereby limiting their clinical applications. Here, we designed monomeric G-quadruplex-based CpG ODNs (G4 CpG ODNs) containing CpG motifs in the central loop region of the G4 structure. The monomeric G4 CpG ODNs were more stable in serum than the linear ODNs. The monomeric G4 CpG ODNs containing two or three CpG motifs induced the production of immunostimulatory cytokines interleukin (IL)-6, IL-12, and interferon (IFN)-beta in mouse macrophage-like RAW264 cells. We also showed that the number of CpG motifs and the number of nucleotides between the CpG motif and G-tracts define the efficacy of the G4 CpG ODNs in activating TLR9. Incubating human peripheral blood mononuclear cells with G4 CpG ODNs promoted IL-6 and IFN-gamma production, confirming their stimulatory effects on human immune cells. Mice given intraperitoneal injections of G4 CpG ODNs produced higher plasma IL-6 compared with injections of linear ODNs. These findings provide further understanding of the parameters governing the immunostimulatory activity of G4 CpG ODNs, thereby providing insights into the rational design of highly potent G4 CpG ODNs for vaccine adjuvants.
2020年09月, 研究論文（学術雑誌）, 共同, 21, 9, 1525-7797, DOI(公開)(r-map), 3644, 3657 - Application of a Glucose Dehydrogenase-Fused with Zinc Finger Protein to Label DNA Aptamers for the Electrochemical Detection of VEGF
Lee, Jinhee; Tatsumi, Atsuro; Tsukakoshi, Kaori; Wilson, Ellie D.; Abe, Koichi; Sode, Koji; Ikebukuro, Kazunori
SENSORS
MDPI
Aptamer-based electrochemical sensors have gained attention in the context of developing a diagnostic biomarker detection method because of their rapid response, miniaturization ability, stability, and design flexibility. In such detection systems, enzymes are often used as labels to amplify the electrochemical signal. We have focused on glucose dehydrogenase (GDH) as a labeling enzyme for electrochemical detection owing to its high enzymatic activity, availability, and well-established electrochemical principle and platform. However, it is difficult and laborious to obtain one to one labeling of a GDH-aptamer complex with conventional chemical conjugation methods. In this study, we used GDH that was genetically fused to a DNA binding protein, i.e., zinc finger protein (ZF). Fused GDH can be attached to an aptamer spontaneously and site specifically in a buffer by exploiting the sequence-specific binding ability of ZF. Using such a fusion protein, we labeled a vascular endothelial growth factor (VEGF)-binding aptamer with GDH and detected the target electrochemically. As a result, upon the addition of glucose, the GDH labeled on the aptamer generated an amperometric signal, and the current response increased dependent on the VEGF concentration. Eventually, the developed electrochemical sensor proved to detect VEGF levels as low as 105 pM, thereby successfully demonstrating the concept of using ZF-fused GDH to enzymatically label aptamers.
2020年07月, 研究論文（学術雑誌）, 共同, 20, 14, DOI(公開)(r-map) - Employment of 1-Methoxy-5-Ethyl Phenazinium Ethyl Sulfate as a Stable Electron Mediator in Flavin Oxidoreductases-Based Sensors
Fitriana, Maya; Loew, Noya; Witarto, Arief Budi; Ikebukuro, Kazunori; Sode, Koji; Tsugawa, Wakako
SENSORS
MDPI
In this paper, a novel electron mediator, 1-methoxy-5-ethyl phenazinium ethyl sulfate (mPES), was introduced as a versatile mediator for disposable enzyme sensor strips, employing representative flavin oxidoreductases, lactate oxidase (LOx), glucose dehydrogenase (GDH), and fructosyl peptide oxidase (FPOx). A disposable lactate enzyme sensor with oxygen insensitive Aerococcus viridans-derived engineered LOx (AvLOx), with A96L mutant as the enzyme, was constructed. The constructed lactate sensor exhibited a high sensitivity (0.73 +/- 0.12 mu A/mM) and wide linear range (0-50 mM lactate), showings that mPES functions as an effective mediator for AvLOx. Employing mPES as mediator allowed this amperometric lactate sensor to be operated at a relatively low potential of +0.2 V to 0 V vs. Ag/AgCl, thus avoiding interference from uric acid and acetaminophen. The lactate sensors were adequately stable for at least 48 days of storage at 25 degrees C. These results indicated that mPES can be replaced with 1-methoxy-5-methyl phenazinium methyl sulfate (mPMS), which we previously reported as the best mediator for AvLOx-based lactate sensors. Furthermore, this study revealed that mPES can be used as an effective electron mediator for the enzyme sensors employing representative flavin oxidoreductases, GDH-based glucose sensors, and FPOx-based hemoglobin A1c (HbA1c) sensors.
2020年05月, 研究論文（学術雑誌）, 共同, 20, 10, DOI(公開)(r-map) - Rational engineering of Aerococcus viridans L-lactate oxidase for the mediator modification to achieve quasi-direct electron transfer type lactate sensor
Hiraka, Kentaro; Kojima, Katsuhiro; Tsugawa, Wakako; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
The L-lactate oxidase (LOx) based lactate sensors are widely used for clinical diagnostics, sports medicine, and food quality control. However, dissolved oxygen interference and electroactive interferent effects are inherent issues of current lactate sensors. In this paper, a quasi-direct electron transfer (quasi-DET) type lactate sensor was developed using rationally engineered Aerococcus viridans LOx (AvLOx) modified with amine-reactive phenazine ethosulfate (PES). Since the modification of wild type AvLOx by PES did not result quasi-DET, engineered AvLOx with additional Lys residue was designed. The additional Lys residue was introduced by substituting residue locating on the surface of AvLOx, and within 20 angstrom of the isoalloxazine ring of FMN. Among several constructed mutants, Ala96Leu/Asn212Lys double mutant showed the highest dye-mediated dehydrogenase activity with negligible oxidase activity, showing quasi-DET properties after PES modification, when the enzyme was immobilized on screen printed carbon electrode. The constructed electrode did not show oxygen interference in cyclic voltammetric analysis and distinct catalytic current with 20 mM L-lactate. The sensor performance of a chronoamperometric L-lactate sensor employing PES modified Ala96Leu/Asn212Lys AvLOx, marked with linear range between 0 and 1 mM, with sensitivity of 13 mu A/mM.cm(2), and a limit of detection of 25 mu M for L-lactate. By applying -200 mV vs. Ag/AgCl, L-lactate could be monitored with negligible interference from 170 mu M ascorbic acid, 1.3 mM acetaminophen, 1.4 mM uric acid or 20 mM glucose. These results indicated that a quasi-DET type lactate sensor was developed that did not suffer from the interference of oxygen and representative electroactive ingredient compounds.
2020年03月01日, 研究論文（学術雑誌）, 共同, 151, 0956-5663, DOI(公開)(r-map) - Ethanol Detection at the Parts per Billion Level with Single-Stranded-DNA-Modified Graphene Field-Effect Transistors
Nozaki, Ryo; Ikuta, Takashi; Ueno, Kinuko; Tsukakoshi, Kaori; Ikebukuro, Kazunori; Maehashi, Kenzo
PHYSICA STATUS SOLIDI B-BASIC SOLID STATE PHYSICS
WILEY-V C H VERLAG GMBH
For realizing the early diagnosis of diseases, detection of gases in exhaled breath in parts per billion (ppb) using compact devices is necessary. Herein, single-stranded-DNA-modified graphene field-effect transistors (FETs) are fabricated and the transfer characteristics are measured. The results reveal that shifts in the transfer characteristics are obtained by introducing ethanol gas at the ppb level, indicating that single-stranded-DNA-modified graphene FETs detect the gas at the ppb level. Moreover, by modifying DNA sequence 1 or sequence 2, the positive and negative voltage shifts of transfer characteristics are observed, respectively. In contrast, the shifts are hardly obtained using DNA sequence 3, which has a rigid conformation for adsorption of molecules. These differences in the behaviors of the transfer characteristics are attributed to the change in DNA conformation when gas is adsorbed. Therefore, single-stranded-DNA-modified graphene FETs have considerable potential to detect various gases at a low concentration depending on the design of DNA sequences, enabling their promising applications in disease diagnosis.
2020年02月, 研究論文（学術雑誌）, 共同, 257, 2, 0370-1972, DOI(公開)(r-map) - G-Quadruplex Structure Improves the Immunostimulatory Effects of CpG Oligonucleotides
Hoshi, Kazuaki; Yamazaki, Tomohiko; Sugiyama, Yuuki; Tsukakoshi, Kaori; Tsugawa, Wakako; Sode, Koji; Ikebukuro, Kazunori
NUCLEIC ACID THERAPEUTICS
MARY ANN LIEBERT, INC
Single-strand oligodeoxynucleotides (ODNs) containing unmethylated cytosine-phosphate-guanine (CpG) are recognized by the toll-like receptor 9, a component of the innate immunity. Therefore, they could act as immunotherapeutic agents. Chemically modified CpG ODNs containing a phosphorothioate backbone instead of phosphodiester (PD) were developed as immunotherapeutic agents resistant to nuclease degradation. However, they cause adverse side effects, and so there is a necessity to generate novel CpG ODNs. In the present study, we designed a nuclease-resistant nonmodified CpG ODN that forms G-quadruplex structures. G-quadruplex formation in CpG ODNs increased nuclease resistance and cellular uptake. The CpG ODNs designed in this study induced interleukin-6 production in a human B lymphocyte cell line and human peripheral blood mononuclear cells. These results indicate that G-quadruplex formation can be used to increase the immunostimulatory activity of CpG ODNs having a natural PD backbone.
2019年08月01日, 研究論文（学術雑誌）, 共同, 29, 4, 2159-3337, DOI(公開)(r-map), 224, 229 - Generation of C5-desoxy analogs of tetrahydroisoquinoline alkaloids exhibiting potent DNA alkylating ability
Tanifuji, Ryo; Tsukakoshi, Kaori; Ikebukuro, Kazunori; Oikawa, Hideaki; Oguri, Hiroki
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS
PERGAMON-ELSEVIER SCIENCE LTD
C5-desoxy analogs of tetrahydroisoquinoline (THIQ) alkaloids were designed and synthesized as hitherto unexplored structural variants for evaluation of their DNA alkylating activities. While chemical synthesis of the C5-desoxy analogs bearing a phenolic hydroxyl group in the A-ring of the saframycins was assumed to be laborious based on semi-synthetic modifications, a chemo-enzymatic approach allowed for concise access to the analogs. The C5-desoxy analog 7 exhibited greater DNA alkylating ability with a wider tolerance for the sequence variations compared to cyanosafracin B. The C5-desoxy A-ring having a C8 phenolic hydroxyl group, and a C1 substituent in the vicinity of the C21 aminonitrile responsible for DNA alkylation, were demonstrated to play pivotal roles in the interaction between the THIQ alkaloids and DNA.
2019年07月15日, 研究論文（学術雑誌）, 共同, 29, 14, 0960-894X, DOI(公開)(r-map), 1807, 1811 - Model studies for isolation of G-quadruplex-forming DNA sequences through a pull-down strategy with macrocyclic polyoxazole
Iida, Keisuke; Tsushima, Yamato; Ma, Yue; Masoud, Shadi Sedghi; Sakuma, Mai; Yokoyama, Tomomi; Yoshida, Wataru; Ikebukuro, Kazunori; Nagasawa, Kazuo
BIOORGANIC & MEDICINAL CHEMISTRY
PERGAMON-ELSEVIER SCIENCE LTD
G-quadruplexes (G4s) are non-B DNA structures present in guanine-rich regions of gene regulatory areas, promoters and CpG islands, but their occurrence and functions remain incompletely understood. Thus, methodology to identify G4 sequences is needed. Here, we describe the synthesis of a novel cyclic hepta-oxazole compound, L1Bio-7OTD (1), bearing a biotin affinity-tag as a tool to pull down G4 structures from mixtures of G4-forming and non G4-forming DNA sequences. We confirmed that it could pull down G4s associated with telomeres, bcl-2 gene, and c-kit gene.
2019年04月15日, 研究論文（学術雑誌）, 共同, 27, 8, 0968-0896, DOI(公開)(r-map), 1742, 1746 - Anti-Idiotype DNA Aptamer Affinity Purification-High-Temperature Reversed-Phase Liquid Chromatography: A Simple, Accurate, and Selective Bioanalysis of Bevacizumab
Yamada, Tomohiro; Saito, Taro; Shimizu, Yutaka; Tsukakoshi, Kaori; Hayashi, Hideki; Mizuno, Hajime; Tsuji, Daiki; Yamamoto, Keisuke; Itoh, Kunihiko; Toyo'oka, Toshimasa; Ikebukuro, Kazunori; Todoroki, Kenichiro
MOLECULES
MDPI
This study presents a simple, accurate, and selective bioanalytical method of bevacizumab detection from plasma samples based on aptamer affinity purification-high-temperature reversed-phased liquid chromatography (HT-RPLC) with fluorescence detection. Bevacizumab in plasma samples was purified using magnetic beads immobilized with an anti-idiotype DNA aptamer for bevacizumab. The purified bevacizumab was separated with HT-RPLC and detected with its native fluorescence. Using aptamer affinity beads, bevacizumab was selectively purified and detected as a single peak in the chromatogram. HT-RPLC achieved good separation for bevacizumab with a sharp peak within 10 min. The calibration curves of the two monoclonal antibodies ranged from 1 to 50 mu g/mL and showed good correlation coefficients (r(2) > 0.999). The limit of detection (LOD) and lower limit of quantification (LLOQ) values for bevacizumab were 0.15 and 0.51 mu g/mL, respectively. The proposed method was successfully applied to the bioanalysis of the plasma samples obtained from the patients with lung cancer and may be extended to plan optimal therapeutic programs and for the evaluation of biological equivalencies in the development of biosimilars.
2019年03月01日, 研究論文（学術雑誌）, 共同, 24, 5, 1420-3049, DOI(公開)(r-map) - High-Throughput Bioanalysis of Bevacizumab in Human Plasma Based on Enzyme-Linked Aptamer Assay Using Anti-Idiotype DNA Aptamer
Yamada, Tomohiro; Saito, Taro; Hill, Yoshia; Shimizu, Yutaka; Tsukakoshi, Kaori; Mizuno, Hajime; Hayashi, Hideki; Ikebukuro, Kazunori; Toyo'oka, Toshimasa; Todoroki, Kenichiro
ANALYTICAL CHEMISTRY
AMER CHEMICAL SOC
We propose a highly selective, sensitive, accurate, and high throughput bioanalysis method for bevacizumab utilizing an anti-idiotype DNA aptamer. With this method, bevacizumab in a plasma sample was reacted in a 96-well plate immobilized with the aptamer and further reacted with a protein A HRP conjugate. The resulting HRP activity was colorimetrically detected using a microplate reader. The calibration curve of bevacizumab ranged from 0.05 to 5.0 mu g/mL, and showed a good correlation coefficient (r(2) = 1.000). The limit of detection was 2.09 ng/mL. We also demonstrated both the possibility of highly sensitive detection using luminol chemiluminescence and the repeated use of affinity plates. The proposed method is applicable for planning optimal therapeutic programs and for an evaluation of the biological equivalencies in the development of biosimilars.
2019年02月19日, 研究論文（学術雑誌）, 共同, 91, 4, 0003-2700, DOI(公開)(r-map), 3125, 3130 - Development of a third-generation glucose sensor based on the open circuit potential for continuous glucose monitoring
Lee, Inyoung; Loew, Noya; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
Continuous glucose monitoring (CGM) systems are most important in the current Type I diabetes care and as component for the development of artificial pancreas systems because the amount of insulin being supplied is calculated based on the CGM results. Therefore, to stably and accurately control the blood glucose level, CGM should be stable and accurate for a long period. We have been engaged in the biomolecular engineering and application of FAD dependent glucose dehydrogenase complex (FADGDH) which is capable of direct electron transfer. In this study, we report the development of the third-generation type open circuit potential (OCP) principle-based glucose sensor with direct electron transfer FADGDH immobilized on gold electrodes using a self-assembled monolayer (SAM). We developed a novel algorithm for OCP-based glucose sensors. By employing this new algorithm, high reproducibility of measurement and sensor preparation were achieved. In addition, the signal was not affected by the presence of acetaminophen and ascorbic acid in the sample solution. The thus optimized third-generation OCP-based glucose sensor could be operated continuously for more than 9 days without significant change in the signal, sensitivity and dynamic range, indicating its potential application for CGM systems.
2019年01月15日, 研究論文（学術雑誌）, 共同, 124, 0956-5663, DOI(公開)(r-map), 216, 223 - Designer fungus FAD glucose dehydrogenase capable of direct electron transfer
Ito, Kohei; Okuda-Shimazaki, Junko; Mori, Kazushige; Kojima, Katsuhiro; Tsugawa, Wakako; Ikebukuro, Kazunori; Lin, Chi-En; La Belle, Jeffrey; Yoshida, Hiromi; Sode, Koji
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
Fungi-derived flavin adenine dinucleotide glucose dehydrogenases (FADGDHs) are currently the most popular and advanced enzymes for self-monitoring of blood glucose sensors; however, the achievement of direct electron transfer (DET) with FADGDHs is difficult. In this study, a designer FADGDH was constructed by fusing Aspergillus flavus derived FADGDH (AfGDH) and a Phanerochaete chrisosporium CDH (PcCDH)-derived heme b-binding cytochrome domain to develop a novel FADGDH that is capable of direct electron transfer with an electrode. A structural prediction suggested that the heme in the CDH may exist in proximity to the FAD of AfGDH if the heme b-binding cytochrome domain is fused to the AfGDH N-terminal region. Spectroscopic observations of recombinantly produced designer FADGDH confirmed the intramolecular electron transfer between FAD and the heme. A decrease in pH and the presence of a divalent cation improved the intramolecular electron transfer. An enzyme electrode with the immobilized designer FADGDH showed an increase in current immediately after the addition of glucose in a glucose concentration-dependent manner, whereas those with wild-type AfGDH did not show an increase in current. Therefore, the designer FADGDH was confirmed to be a novel GDH that possesses electrode DET ability. The difference in the surface electrostatic potentials of AfGDH and the catalytic domain of PcCDH might be why their intramolecular electron transfer ability is inferior to that of CDH. These relevant and consistent findings provide us with a novel strategic approach for the improvement of the DET properties of designer FADGDH. (241 words)
2019年01月01日, 研究論文（学術雑誌）, 共同, 123, 0956-5663, DOI(公開)(r-map), 114, 123 - Selection and Characterization of DNA Aptamers Against FokI Nuclease Domain.
Nishio M, Yamagishi A, Tsukakoshi K, Kato Y, Nakamura C, Ikebukuro K.
Methods Mol Biol.
2018年, 研究論文（学術雑誌）, 共同, 1867:, DOI(公開)(r-map), 165, 174 - Synthesis of a hemin-containing copolymer as a novel immunostimulator that induces IFN-gamma production
Hoshi, Kazuaki; Yamazaki, Tomohiko; Yoshikawa, C. Hiaki; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji
INTERNATIONAL JOURNAL OF NANOMEDICINE
DOVE MEDICAL PRESS LTD
Background: Hemozoin, a chemical analog of a malarial pigment, is a crystal composed of heme dimers that can act as a potent Th1-type adjuvant, which strongly induces antibody production. However, the clinical applications of malarial hemozoin have limitations due to biosafety concerns and difficulties in the manufacturing process. Based on the premise that an analog of the heme polymer might display immunostimulatory effects, a hemin-containing polymer was developed as a novel immunostimulator. Materials and methods: To synthesize the copolymer containing hemin and N-isopropylacrylamide (NIPAM), this study employed a conventional radical polymerization method using 2,2'-azodiisobutyronitrile as the radical initiator; the synthesized copolymer was designated as NIPAM-hemin. Results: NIPAM-hemin was soluble and showed no cytotoxicity in vitro. The NIPAM-hemin copolymer induced the production of interferon (IFN)-gamma and interleukin (IL)-6 from peripheral blood mononuclear cells, although hemin and the NIPAM monomer individually did not induce the production of any cytokines. The production of IFN-gamma induced by NIPAM-hemin was independent of toll-like receptor 9 and the NLRP3 inflammasome pathway. Conclusion: Given that NIPAM-hemin induced IL-6 and IFN-gamma production in immune cells without any cytotoxic effects, NIPAM-hemin has potential therapeutic applications as a Th1-type adjuvant.
2018年, 研究論文（学術雑誌）, 共同, 13, 1178-2013, DOI(公開)(r-map), 4461, 4472 - Improving the induction fold of riboregulators for cyanobacteria
Sakamoto, Ippei; Abe, Koichi; Kawai, Sumiya; Tsukakoshi, Kaori; Sakai, Yuta; Sode, Koji; Ikebukuro, Kazunori
RNA BIOLOGY
TAYLOR & FRANCIS INC
Cyanobacteria are ideal cellular factories for biochemical production because of their ability to fix CO2 by photosynthesis and convert this molecule into biochemicals. Previously, we engineered a riboregulator that enables post-transcriptional gene regulation in the cyanobacterium Synechocystis sp. PCC 6803. Here, we improved the riboregulator by designing two RNA species, taRNA and crRNA, to enhance its induction fold. We inserted nucleotides into the crRNA loop to enhance intermolecular hybridization and successfully improved its induction fold. The engineered riboregulator exhibited a higher induction fold than the previously engineered riboregulator in both Escherichia coli and Synechocystis sp. PCC 6803. This improved riboregulator can be used to control gene expression over a wide dynamic range in cyanobacteria.
2018年, 研究論文（学術雑誌）, 共同, 15, 3, 1547-6286, DOI(公開)(r-map), 353, 358 - Esterification of PQQ Enhances Blood-Brain Barrier Permeability and Inhibitory Activity against Amyloidogenic Protein Fibril Formation
Tsukakoshi, Kaori; Yoshida, Wataru; Kobayashi, Masaki; Kobayashi, Natsuki; Kim, Jihoon; Kaku, Toshisuke; Iguchi, Toshitsugu; Nagasawa, Kazuo; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji
ACS CHEMICAL NEUROSCIENCE
AMER CHEMICAL SOC
Several neurodegenerative diseases have a common pathophysiology where selective damage to neurons results from the accumulation of amyloid oligomer proteins formed via fibrilization. Considering that the formation of amyloid oligomers leads to cytotoxicity, the development of chemical compounds that are able to effectively cross the blood-brain barrier (BBB) and inhibit this conversion to oligomers and/or fibrils is essential for neurodegenerative disease therapy. We previously reported that pyrroloquinoline quinone (PQQ) prevented aggregation and fibrillation of alpha-synuclein, amyloid beta(1-42) (A beta(1-42)), and mouse prion protein. To develop a novel drug against neurodegenerative diseases based on PQQ, it is necessary to improve the insufficient BBB permeability of PQQ. Here, we show that an esterified compound of PQQ, PQQ-trimethylester (PQQ-TME), has twice the BBB permeability than PQQ in vitro. Moreover, PQQ-TME exhibited greater inhibitory activity against fibrillation of alpha-synuclein, A beta(1-42), and prion protein. These results indicated that esterification of PQQ could be a useful approach in developing a novel PQQ-based amyloid inhibitor.
2018年12月, 研究論文（学術雑誌）, 共同, 9, 12, 1948-7193, DOI(公開)(r-map), 2898, 2903 - Riboregulator elements as tools to engineer gene expression in cyanobacteria
Ueno, Kinuko; Tsukakoshi, Kaori; Ikebukuro, Kazunori
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
SPRINGER
Cyanobacteria are an ideal host for biofuel production. Although efforts have been made to genetically engineer cyanobacteria for efficient production of biofuels and other important chemicals, the tools that can be applied to cyanobacteria are still limited. A new gene regulation tool, riboregulator, has been examined for application in cyanobacteria. A riboregulator is a nature-inspired RNA tool, which is composed of two artificially designed RNA fragments. Owing to its high specificity and efficacy, it is suitable for metabolic engineering in cyanobacteria, and several studies have been done to optimize and improve the function of the riboregulator. In this review, we focus on the recent improvements made to riboregulators and compare them with other RNA-mediated gene regulation tools developed in cyanobacteria to investigate future applications of riboregulators.
2018年09月, 研究論文（学術雑誌）, 共同, 102, 18, 0175-7598, DOI(公開)(r-map), 7717, 7723 - Self-powered, wireless continuous glucose sensing system based direct electron transfer
Lee, Inyoung; Loew, Noya; Tsugawa, Wakako; Ikebukuro, Kazunori; Sode, Koji
ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY
AMER CHEMICAL SOC
2018年08月19日, 研究論文（学術雑誌）, 共同, 256, 0065-7727, DOI(公開)(r-map) - Development of an electrochemical detection system for CpG methylation of human genome using methyl CpG binding domain and zinc finger protein
Lee, Jinhee; Yoshida, Wataru; Hiraoka, Daisuke; Tatsumi, Atsuro; Abe, Koichi; Nakabayashi, Kazuhiko; Wakeda, Hironobu; Hata, Kenichiro; Marquete, Christophe; Blum, Loic; Sode, Koji; Ikebukuro, Kazunori
ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY
AMER CHEMICAL SOC
2018年08月19日, 研究論文（学術雑誌）, 共同, 256, 0065-7727, DOI(公開)(r-map) - Smart aptamers forming G-quadruplex: Structural and functional change of aptamers forming G-quadruplex in response to surrounding conditions and its regulation with the ligands
Tsukakoshi, Kaori; Nishio, Maui; Sasaki, Ikkei; Ma, Yue; Nagasawa, Kazuo; Kato, Yoshio; Nakamura, Chikashi; Sode, Koji; Ikebukuro, Kazunori
ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY
AMER CHEMICAL SOC
2018年08月19日, 研究論文（学術雑誌）, 共同, 256, 0065-7727, DOI(公開)(r-map) - Riboregulator elements as tools to engineer gene expression in cyanobacteria.
Ueno, Kinuko; Tsukakoshi, Kaori; Ikebukuro, Kazunori
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
SPRINGER
Cyanobacteria are one of the most attractive hosts for biofuel production; however, genetic approaches to regulate specific chromosomal genes in cyanobacteria remain limited. With the aim of developing a novel method to regulate chromosomal gene expression in cyanobacteria, we focused on riboregulatory technology. Riboregulators are composed of two RNA fragments whose interaction leads to target gene regulation with high specificity. In this study, we inserted a riboregulator sequence upstream of the chromosomal gene encoding AbrB-like transcriptional regulator, cyAbrB2, to investigate the utility of this tool. The inserted riboregulator was able to regulate cyabrB2 gene expression, with a high ON-OFF ratio up to approximately 50-fold. The transcription levels of several genes for which cyAbrB2 acts as a transcriptional upregulator were also decreased. Further, the cyAbrB2 expression-repressed mutant showed high glycogen accumulation, equivalent to that in the cyabrB2 deletion mutant (Delta cyabrB2). Phenotypic similarities between the cyabrB2 expression-repressed mutant and the Delta cyabrB2 mutant suggest that the riboregulator can potentially be used as a new chromosomal gene regulation tool in cyanobacteria.
2018年07月13日, 研究論文（学術雑誌）, 共同, 102(18):, 18, DOI(公開)(r-map), 7717, 7723 - CpG Methylation Changes G-Quadruplex Structures Derived from Gene Promoters and Interaction with VEGF and SP1
Tsukakoshi, Kaori; Saito, Shiori; Yoshida, Wataru; Goto, Shinichi; Ikebukuro, Kazunori
MOLECULES
MDPI
G-quadruplex (G4) is a DNA/RNA conformation that consists of two or more G-tetrads resulting from four-guanine bases connected by Hoogsteen-type hydrogen bonds, which is often found in the telomeres of chromatin, as well as in the promoter regions of genes. The function of G4 in the genomic DNA is being elucidated and some G4-protein interactions have been reported; these are believed to play a role in vital cellular functions. In this study, we focused on CpG methylation, a well-known epigenetic modification of the genomic DNA, especially found in the promoter regions. Although many G4-forming sequences within the genomic DNA harbor CpG sites, the relationship between CpG methylation and the binding properties of associated proteins remains unclear. We demonstrated that the binding ability of vascular endothelial growth factor (VEGF) G4 DNA to VEGF165 protein was significantly decreased by CpG methylation. We identified the binding activity of G4 DNA oligonucleotides derived from gene promoter regions to SP1, a transcription factor that interacts with a G4-forming DNA and is also altered by CpG methylation. The effect of methylation on binding affinity was accompanied by changes in G4 structure and/or topology. Therefore, this study suggested that CpG methylation might be involved in protein binding to G4-forming DNA segments for purposes of transcriptional regulation.
2018年04月, 研究論文（学術雑誌）, 共同, 23, 4, 1420-3049, DOI(公開)(r-map) - Improving the induction fold of riboregulators for cyanobacteria,
Sakamoto I, Abe K, Kawai S, Tsukakoshi K, Sakai Y, Sode K, Ikebukuro
RNA Biol
2018年02月, 研究論文（学術雑誌）, 共同, 1-6, doi: 10.1080/15476286.2017.1422470. [Epub ahead of print] - Identification of G-quadruplex clusters by high-throughput sequencing of whole-genome amplified products with a G-quadruplex ligand
Yoshida, Wataru; Saikyo, Hiroki; Nakabayashi, Kazuhiko; Yoshioka, Hitomi; Bay, Daniyah Habiballah; Iida, Keisuke; Kawai, Tomoko; Hata, Kenichiro; Ikebukuros, Kazunori; Nagasawas, Kazuo; Karube, Isao
SCIENTIFIC REPORTS
NATURE PUBLISHING GROUP
G-quadruplex (G4) is a DNA secondary structure that has been found to play regulatory roles in the genome. The identification of G4-forming sequences is important to study the specific structure-function relationships of such regions. In the present study, we developed a method for identification of G4 clusters on genomic DNA by high-throughput sequencing of genomic DNA amplified via whole-genome amplification (WGA) in the presence of a G4 ligand. The G4 ligand specifically bound to G4 structures on genomic DNA; thus, DNA polymerase was arrested on the G4 structures stabilised by G4 ligand. We utilised the telomestatin derivative L1H1-7OTD as a G4 ligand and demonstrated that the efficiency of amplification of the G4 cluster regions was lower than that of the non-G4-forming regions. By high-throughput sequencing of the WGA products, 9,651 G4 clusters were identified on human genomic DNA. Among these clusters, 3,766 G4 clusters contained at least one transcriptional start site, suggesting that genes are regulated by G4 clusters rather than by one G4 structure.
2018年02月15日, 研究論文（学術雑誌）, 共同, 8, 2045-2322, DOI(公開)(r-map) - Development of aptamers against unpurified proteins
Goto, Shinichi; Tsukakoshi, Kaori; Ikebukuro, Kazunori
BIOTECHNOLOGY AND BIOENGINEERING
WILEY
SELEX (Systematic Evolution of Ligands by EXponential enrichment) has been widely used for the generation of aptamers against target proteins. However, its requirement for pure target proteins remains a major problem in aptamer selection, as procedures for protein purification from crude bio-samples are not only complicated but also time and labor consuming. This is because native proteins can be found in a large number of diverse forms because of posttranslational modifications and their complicated molecular conformations. Moreover, several proteins are difficult to purify owing to their chemical fragility and/or rarity in native samples. An alternative route is the use of recombinant proteins for aptamer selection, because they are homogenous and easily purified. However, aptamers generated against recombinant proteins produced in prokaryotic cells may not interact with the same proteins expressed in eukaryotic cells because of posttranslational modifications. Moreover, to date recombinant proteins have been constructed for only a fraction of proteins expressed in the human body. Therefore, the demand for advanced SELEX methods not relying on complicated purification processes from native samples or recombinant proteins is growing. This review article describes several such techniques that allow researchers to directly develop an aptamer from various unpurified samples, such as whole cells, tissues, serum, and cell lysates. The key advantages of advanced SELEX are that it does not require a purification process from a crude bio-sample, maintains the functional states of target proteins, and facilitates the development of aptamers against unidentified and uncharacterized proteins in unpurified biological samples.
2017年12月, 研究論文（学術雑誌）, 共同, 114, 12, 0006-3592, DOI(公開)(r-map), 2706, 2716 - Applying a riboregulator as a new chromosomal gene regulation tool for higher glycogen production in Synechocystis sp. PCC 6803,
3. Ueno K, Sakai Y, Shono C, Sakamoto I, Tsukakoshi K, Hihara Y, Sode K, Ikebukuro K,
Appl Microbiol Biotechnol
2017年11月, 研究論文（学術雑誌）, 共同, 101, 23-24, 8465, 8474. doi: 10.1007/s00253-017-8570-4 - Development of HGF-binding aptamers with the combination of G4 promoter-derived aptamer selection and in silico maturation
Yokoyama, Tomomi; Tsukakoshi, Kaori; Yoshida, Wataru; Saito, Taiki; Teramoto, Kentaro; Savory, Nasa; Abe, Koichi; Ikebukuro, Kazunori
BIOTECHNOLOGY AND BIOENGINEERING
WILEY
We describe the selection of aptamers based on bioinformatics-based approaches without Systematic Evolution of Ligands by EXponential enrichment (SELEX). SELEX is a potent method; however, it is time intensive and the PCR-amplification step, which is essential step for SELEX, leads to the loss of good aptamers. We have developed an aptamer-screening method, G4 promoter-derived aptamer selection (G4PAS), and an aptamer-improving method, in silico maturation (ISM). They are based on in silico sequence selection and computer assisted directed evolution, respectively. In this study, we succeeded in identifying new aptamers against hepatocyte growth factor (HGF) by G4PAS as well as improving the specificity of the HGF aptamers by ISM. Using ISM improved the specificity of the aptamer for HGF by up to 45-fold in comparison with the original aptamer. These methods enable easy and efficient identification of good aptamers, and the combination of G4PAS with ISM can thus serve as a potent approach for aptamer identification. Biotechnol. Bioeng. 2017;114: 2196-2203. (c) 2017 Wiley Periodicals, Inc.
2017年10月, 研究論文（学術雑誌）, 共同, 114, 10, 0006-3592, DOI(公開)(r-map), 2196, 2203 - Development of an electrochemical detection system for measuring DNA methylation levels using methyl CpG-binding protein and glucose dehydrogenase-fused zinc finger protein
Lee, Jinhee; Yoshida, Wataru; Abe, Koichi; Nakabayashi, Kazuhiko; Wakeda, Hironobu; Hata, Kenichiro; Marquette, Christophe A.; Blum, Loic J.; Sode, Koji; Ikebukuro, Kazunori
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
DNA methylation level at a certain gene region is considered as a new type of biomarker for diagnosis and its miniaturized and rapid detection system is required for diagnosis. Here we have developed a simple electrochemical detection system for DNA methylation using methyl CpG-binding domain (MBD) and a glucose dehydrogenase (GDH)-fused zinc finger protein. This analytical system consists of three steps: (1) methylated DNA collection by MBD, (2) PCR amplification of a target genomic region among collected methylated DNA, and (3) electrochemical detection of the PCR products using a GDH-fused zinc finger protein. With this system, we have successfully measured the methylation levels at the promoter region of the androgen receptor gene in 106 copies of genomic DNA extracted from PC3 and TSU-PR1 cancer cell lines. Since no sequence analysis or enzymatic digestion is required for this detection system, DNA methylation levels can be measured within 3 h with a simple procedure.
2017年07月, 研究論文（国際会議プロシーディングス）, 共同, 93, 0956-5663, DOI(公開)(r-map), 118, 123 - DNA aptamers against FokI nuclease domain for genome editing applications
Nishio, Maui; Matsumoto, Daisuke; Kato, Yoshio; Abe, Koichi; Lee, Jinhee; Tsukakoshi, Kaori; Yamagishi, Ayana; Nakamura, Chikashi; Ikebukuro, Kazunori
BIOSENSORS & BIOELECTRONICS
ELSEVIER ADVANCED TECHNOLOGY
Genome editing with site-specific nucleases (SSNs) can modify only the target gene and may be effective for gene therapy. The main limitation of genome editing for clinical use is off-target effects; excess SSN5 in the cells and their longevity can contribute to off-target effects. Therefore, a controlled delivery system for SSN5 is necessary. Fold nuclease domain (FokI) is a common DNA cleavage domain in zinc finger nuclease (ZFN) and transcription activator-like effector nuclease. Previously, we reported a zinc finger protein delivery system that combined aptamer-fused, double-strand oligonucleotides and nanoneedles. Here, we report the development of DNA aptamers that bind to the target molecules, with high affinity and specificity to the FokI. DNA aptamers were selected in six rounds of systematic evolution of ligands by exponential enrichment. Aptamers F6#8 and #71, which showed high binding affinity to Fokl (K-d=82 nM, 74 nM each), showed resistance to nuclease activity itself and did not inhibit nuclease activity. We immobilized the ZFN-fused GFP to nanoneedles through these aptamers and inserted the nanoneedles into HEK293 cells. We observed the release of ZFN-fused GFP from the nanoneedles in the presence of cells. Therefore, these aptamers are useful for genome editing applications such as controlled delivery of SSNs.
2017年07月, 研究論文（国際会議プロシーディングス）, 共同, 93, 0956-5663, DOI(公開)(r-map), 26, 31 - , Development of HGF-binding aptamers with the combination of G4 promoter-derived aptamer selection and in silico maturation
Yokoyama T, Tsukakoshi K, Yoshida W, Saito T, Teramoto K, Savory N, Abe K, Ikebukuro K
Biotechnol Bioeng
2017年07月, 共同, 114, 10, 2196, 2203. doi: 10.1002/bit.26354. - Pipette tip biosensors for bacterial double-stranded DNA using bioluminescence induced by zinc finger luciferase
Takano, Eri; Shimura, Nobuaki; Akiba, Takeshi; Kitayama, Yukiya; Sunayama, Hirobumi; Abe, Koichi; Ikebukuro, Kazunori; Takeuchi, Toshifumi
MICROCHIMICA ACTA
SPRINGER WIEN
The authors describe a pipette type of biosensor for detecting target genes and using a zinc finger protein fused to luciferase (ZF luciferase). The ZF protein binds to a specific DNA sequence, and the target double-stranded (ds) DNA can be detected by monitoring the enzymatic activity of ZF luciferase. A small avidin-immobilized reaction plate is placed on a plastic pipette tip (referred to as Biologi tip). The dsDNA detection procedures are carried out by using a programmable dispensing robot equipped with a photodetector. These procedures include (a) the aspiration of an analyte to capture the biotinylated target dsDNA (a product of a polymerase chain reaction) on the small reaction plate inside the pipette tip, (b) the introduction of ZF luciferase and luciferin into the pipette tip, and (c) migration of the pipette tip to the detection port to measure bioluminescence on the small reaction plate. The emission originating from luciferase activity is observed on the reaction plate containing immobilized biotin-tagged target dsDNA, whereas plates containing non-target or biotinylated single-stranded DNA only do not yield a signal. The intensity of emission increases proportionally to the concentration of dsDNA, and the detection limit of the target dsDNA is as low as 62 pM. An actual genomic DNA sample from Escherichia coli O157 was successfully detected by this automatic analyzer using the Biologi tip equipped with a reaction plate. This indicates that this system has a large potential for practical applications, including in particular point-of-care analyses in hygiene control, food safety testing, and clinical diagnosis.
2017年06月, 研究論文（学術雑誌）, 共同, 184, 6, 0026-3672, DOI(公開)(r-map), 1595, 1601 - Identification of G-quadruplex structures that possess transcriptional regulating functions in the Dele and Cdc6 CpG islands
Bay, Daniyah H.; Busch, Annika; Lisdat, Fred; Iida, Keisuke; Ikebukuro, Kazunori; Nagasawa, Kazuo; Karube, Isao; Yoshida, Wataru
BMC MOLECULAR BIOLOGY
BIOMED CENTRAL LTD
Background: G-quadruplex is a DNA secondary structure that has been shown to play an important role in biological systems. In a previous study, we identified 1998 G-quadruplex-forming sequences using a mouse CpG islands DNA microarray with a fluorescent-labeled G-quadruplex ligand. Among these putative G-quadruplex-forming sequences, G-quadruplex formation was verified for 10 randomly selected sequences by CD spectroscopy and DMS footprinting analysis. In this study, the biological function of the 10 G-quadruplex-forming sequences in the transcriptional regulation has been analyzed using a reporter assay. Results: When G-quadruplex-forming sequences from the Dele and Cdc6 genes have been cloned in reporter vectors carrying a minimal promoter and the luciferase gene, luciferase expression is activated. This has also been detected in experiments applying a promoterless reporter vector. Mutational analysis reveals that guanine bases, which form the G-tetrads, are important in the activation. In addition, the activation has been found to decrease by the telomestatin derivative L1H1-7OTD which can bind to the G-quadruplex DNA. When Dele and Cdc6 CpG islands, containing the G-quadruplex-forming sequence, have been cloned in the promoterless reporter vector, the luciferase expression is activated. Mutational analysis reveals that the expression level is decreased by mutation on Dele G-quadruplex; however, increased by mutation on Cdc6 G-quadruplex. Conclusion: Dele and Cdc6 G-quadruplex formation is significant in the transcriptional regulation. Dele and Cdc6 G-quadruplex DNA alone possess enhancer and promotor function. When studied in more complex CpG islands Dele G-quadruplex also demonstrates promotor activity, whereas Cdc6 G-quadruplex may possess a dual function of transcriptional regulation.
2017年06月, 研究論文（学術雑誌）, 共同, 18, 1471-2199, DOI(公開)(r-map) - , Pipette tip biosensors for bacterial double-stranded DNA using bioluminescence induced by zinc finger luciferase
9. Takano E, Shimura N, Akiba T, Kitayama Y, Sunayama H, Abe K, Ikebukuro K, Takeuchi T
Microchimica Acta
2017年06月, 共同, 184, 6, 1595, 1601. - Detection of DNA methylation of G-quadruplex and i-motif-forming sequences by measuring the initial elongation efficiency of polymerase chain reaction
Yoshida W, Yoshioka H, Bay DH, Iida K, Ikebukuro K, Nagasawa K, Karube I., ‘
Anal Chem
2016年09月, 研究論文（学術雑誌）, 共同, 88, 14, 7101, 7106. - Structural regulation by a G-quadruplex ligand increases binding abilities of G-quadruplex-forming aptamers
Tsukakoshi K, Ikuta Y, Abe K, Yoshida W, Iida K, Ma Y, Nagasawa K, Sode K, Ikebukuro K.,
Chem Commun (Camb). 2016,
2016年09月, 研究論文（学術雑誌）, 共同, 52, 85, 12646, 12649. - Sensitive and Homogeneous Detection System with Aptamer-Based Biosensor,
Tsukakoshi K, Ikebukuro K,
Sens. Mater.
2016年09月, 共同, 28, 3, 1083, 1089. - Methods for Improving Aptamer Binding Affinity, Molecules,
Hasegawa H, Savory N, Abe K, Ikebukuro K.
Molecules
2016年03月, 共同, 21, 4, E421. - ATP-mediated Release of a DNA-binding Protein from a Silicon Nanoneedle Array
1. Matsumoto, D, Nishio, M, Kato, Y, Yoshida, W, Abe, K, Fukazawa, K, Ishihara, K, Iwata, F, Ikebukuro, K, Nakamura, C,
Electrochemistry
2016年03月, 研究論文（学術雑誌）, 共同, 84, 5, 305, 307. - アプタマーを用いた機能性電極
李ジンヒ、池袋一典
Electrochemistry
2015年12月, 共同, 2015, 12 - Inhibition of an allergen–antibody reaction related to Japanese cedar pollinosis using DNA sptamers sgainst the Cry j 2 allergen, 2015,
Kazumasa Ogihara, Nasa Savory, Koichi Abe, Wataru Yoshida, Mitsuru Arakawa, Masahiko Asahi, Seika Kamohara, and Kazunori Ikebukuro,
Nucleic acid therapeutics,
2015年12月, 研究論文（学術雑誌）, 共同, 212, 57, 99, 105. - DNA detection technology using zinc finger protein,
Kumagai T., Abe K., Yoshida W., Ikebukuro K.,
J. Microb. Biochem. Technol., in press
2015年11月, 共同, in press - Inhibition of an allergen–antibody reaction related to Japanese cedar pollinosis using DNA sptamers sgainst the Cry j 2 allergen, 2015,
K1. Saito T, Yoshida W, Yokoyama T, Abe K, Ikebukuro K, Identification of RNA Oligonucleotides Binding to Several Proteins from Potential G-Quadruplex Forming Regions in Transcribed Pre-mRNA,
Molecules
2015年11月, 研究論文（学術雑誌）, 共同, 20, 11, 20832, 20840. - Improvement of the VEGF binding ability of DNA aptamers through in silico maturation and multimerization strategy
Fukaya T, Abe K, Savory N, Tsukakoshi K, Yoshida W, Ferri S, Sode K, Ikebukuro K
J Biotechnol.
2015年10月, 研究論文（学術雑誌）, 共同, 212, 57, 99, 105. - Scaffold-fused riboregulators for enhanced gene activation in Synechocystis sp. PCC 6803,
Sakai Y, Abe K, Nakashima S, Ellinger JJ, Ferri S, Sode K, Ikebukuro K.
Microbiologyopen
2015年08月, 研究論文（学術雑誌）, 共同, 4, 4, 4881, 4884. - Enzyme linking to DNA aptamers via a zinc finger as a bridge, 2015,
Abe K, Murakami Y, Tatsumi A, Sumida K, Kezuka A, Fukaya T, Kumagai T, Osawa Y, Sode K, Ikebukuro K.,
Chem Commun (Camb)
2015年07月, 研究論文（学術雑誌）, 共同, 51, 57, 11467, 1149. - Development of an automated direct blotting electrophoresis system for bioanalytical applications
Goto S., Savory N , Abe K, Kinoshita H, Ikebukuro K.,
Analytical Methods
2015年01月, 研究論文（学術雑誌）, 共同, 7, 12, 4881, 4884. - DNA aptamers against the Cry j 2 allergen of Japanese cedar pollen for biosensing applications.
Ogihara K, Savory N, Abe K, Yoshida W, Asahi M, Kamohara S, Ikebukuro K
Biosens Bioelectron
2015年01月, 研究論文（学術雑誌）, 共同, 63, 159, 65 - Selection of DNA aptamers against uropathogenic Escherichia coli NSM59 by quantitative PCR controlled Cell-SELEX
Savory N, Nzakizwanayo J, Abe K, Yoshida W, Ferri S, Dedi C, Jones BV, Ikebukuro K.
J Microbiol Methods.
2014年09月, 研究論文（学術雑誌）, 共同, 104, 2, 94, 100. - The development of an autonomous self-powered bio-sensing actuator
Hanashi, Takuya; Yamazaki, Tomohiko; Tanaka, Hiroshi, Ikebukuro, K, Tsugawa, W, Sode, K,
Sensors and Actuators B-Chemical
2014年06月, 研究論文（学術雑誌）, 共同, 196, 2, 429, 433 - Electrochemical detection of pathogenic bacteria by using a glucose dehydrogenase fused zinc finger protein
Lee, Jinhee, Tatsumi, Atsuro, Abe, Koichi, Yoshida, W . Sode, K, Ikebukuro, K, Electrochemical detection of pathogenic bacteria by using a glucose dehydrogenase fused zinc finger protein, Abe K, Miyake K, Nakamura M, Kojima K, Ferri S, Ikebukuro K, Sode K. Abe K, Miyake K, Nakamura M, Kojima K, Ferri S, Ikebukuro K, Sode K
Analytical Methods,
2014年03月, 研究論文（学術雑誌）, 共同, 6, 14, 4991, 4994 - Electrochemical biosensors using aptamers for theranostics.
Emerging techniques employed in aptamer-based diagnostic tests. Yoshida W, Abe K, Ikebukuro K
Expert Rev Mol Diagn.
2014年03月, 共同, 14, 2, 143, 51 - Improving the gene-regulation ability of small RNAs by scaffold engineering in Escherichia coli.
Engineering of a green-light inducible gene expression system in Synechocystis sp. PCC6803. Abe K, Miyake K, Nakamura M, Kojima K, Ferri S, Ikebukuro K, Sode K. Abe K, Miyake K, Nakamura M, Kojima K, Ferri S, Ikebukuro K, Sode K
Microb Biotechnol
2014年03月, 研究論文（学術雑誌）, 共同, 7, 2, 177, 83 - Improving the gene-regulation ability of small RNAs by scaffold engineering in Escherichia coli.
Sakai Y, Abe K, Nakashima S, Yoshida W, Ferri S, Sode K, Ikebukuro K.
ACS Synth Biol
2014年03月, 研究論文（学術雑誌）, 共同, 3, 3, 152, 62 - Simultaneous improvement of specificity and affinity of aptamers against Streptococcus mutans by in silico maturation for biosensor development
10. Savory N, Takahashi Y, Tsukakoshi K, Hasegawa H, Takase M, Abe K, Yoshida W, Ferri S, Kumazawa S, Sode K, Ikebukuro K
Biotechnol Bioeng
2014年03月, 研究論文（学術雑誌）, 共同, 111, 3, 454, 6 - Vascular Endothelial Growth Factor (VEGF) Detection Using an Aptamer and PNA-Based Bound/Free Separation System
Mita, Chifuku, Abe, Koichi; Fukaya, Takahiro, Ikebukuro K.,
Materials
2014年02月, 研究論文（学術雑誌）, 共同, 7, 2, 1046, 1054 - Design of riboregulators for control of cyanobacterial (Synechocystis) protein expression
Abe K, Sakai Y, Nakashima S, Araki M, Yoshida W, Sode K, Ikebukuro K
Biotechnol Lett
2014年02月, 研究論文（学術雑誌）, 共同, 36, 2, 287, 94 - Electrochemical biosensors using aptamers for theranostics.
6. Abe K, Yoshida W, Ikebukuro K
Adv Biochem Eng Biotechnol. 2014; Review
2014年01月, 共同, 140:, 183, 202 - Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase
Yoshida W, Kezuka A, Murakami Y, Lee J, Abe K, Motoki H, Matsuo T, Shimura N, Noda M, Igimi S, Ikebukuro K.
Anal Chim Acta
2013年11月, 研究論文（学術雑誌）, 共同, 801, 78, 83 - Fluorescent-ligand-mediated screening of G-quadruplex structures using a DNA microarray
Iida, K. Takahiro Nakamura, Wataru Yoshida, Masayuki Tera, Kazuhiko Nakabayashi, Kenichiro Hata, Kazunori Ikebukuro,* and Kazuo Nagasawa*
Angew. Chem. Int. Ed.(inside cover)
2013年11月, 研究論文（学術雑誌）, 共同, 52, 46, 12052, 12055 - Two-dimensional electrophoresis-based selection of aptamers against an unidentified protein in a tissue sample
Savory, Nasa, Goto, Shinichi, Yoshida, Wataru, Unuma, Y, Nakamura, M , Abe, K , Ferri, S, Ikebukuro, K
Analytical letters
2013年10月, 研究論文（学術雑誌）, 共同, 46, 18, 2954, 2963 - In silico maturation of binding-specificity of DNA aptamers against Proteus mirabilis
Savory N, Lednor D, Tsukakoshi K, Abe K, Yoshida W, Ferri S, Jones BV, Ikebukuro K
Biotechnol Bioeng
2013年10月, 研究論文（学術雑誌）, 共同, 110, 10, 2573, 2580 - Screening of Peptide ligands for pyrroloquinoline quinone glucose dehydrogenase using antagonistic template-based biopanning
Abe K, Yoshida W, Terada K, Yagi-Ishii Y, Ferri S, Ikebukuro K, Sode K.,
Int J Mol Sci
2013年08月, 研究論文（学術雑誌）, 共同, 14, 12, 23244, 56 - Detection of histone modification by chromatin immunoprecipitation combined zinc finger luciferase-based bioluminescence resonance energy transfer assay
Yoshida W, Kezuka A, Abe K, Wakeda H, Nakabayashi K, Hata K, Ikebukuro K
Anal Chem. 2013 Jul 2;
2013年07月, 研究論文（学術雑誌）, 共同, 85, 13, 6485, 90 - An Optical Biosensing System Based on Interference-Enhanced Reflection with Aptameric Enzyme Subunits of Thrombin
W. Yoshida and K. Ikebukuro
Analytical letters
2013年02月, 研究論文（学術雑誌）, 共同, 46, 2, 242, 249 - Rapid cytotoxicity screening platform for amyloid inhibitors using a membrane-potential sensitive fluorescent probe
Kim J, Sasaki Y, Yoshida W, Kobayashi N, Veloso AJ, Kerman K, Ikebukuro K, Sode K.
Anal Chem. 2013
2013年01月, 研究論文（学術雑誌）, 共同, 85, 1, 185, 92 - Aptamer selection based on g4-forming promoter region
Yoshida W, Saito T, Yokoyama T, Ferri S, Ikebukuro K. (2013)
PLoS One
2013年01月, 研究論文（学術雑誌）, 共同, 8, 6, e65497. - Affinity improvement of a VEGF aptamer by in silico maturation for a sensitive VEGF-detection system
15. Yoshihiko Nonaka, Wataru Yoshida, Koichi Abe, Stefano Ferri, Holger Schulze, Till T. Bachmann and Ikebukuro K
Analytical Chemistry
2013年01月, 研究論文（学術雑誌）, 共同, 85, 2, 1132, 1137 - アミロイド形成蛋白質に結合する核酸リガンド，DNAアプタマーの開発の現状：アミロイドオリゴマー結合アプタマーの同定
塚越かおり、阿部公一、早出広司、池袋一典
Dementia Japan
2012年09月, 共同, 26, 3 - Selection of DNA aptamers that recognize α-synuclein oligomer using a competitive screening method,
Kaori Tsukakoshi, Koichi Abe, Koji Sode, and Kazunori Ikebukuro
Analytical Chemistry
2012年07月, 研究論文（学術雑誌）, 共同, 84, 13, 5542, 5547 - BioLC-Oscillator: A Self-Powered Wireless Glucose-Sensing System with the Glucose Dependent Resonance Frequency
Takuya Hanash, Tomohiko Yamazaki, Wakako Tsugawa, Kazunori Ikebukuro, Koji Sode
Electrochemistry
2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 367, 370 - Electrochemical SNP Detection Using Glucose Dehydrogenase,
Koichi Abe, Chiharu Kiyohara, Yoko Saito, Koji Sode, Kazunori Ikebukuro,
Electrochemistry
2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 345, 347 - Electrochemical Detection of Vascular Endothelial Growth Factor by an Aptamer-Based Bound/Free Separation System,
Koichi Abe, Hijiri Hasegawa, Kazunori Ikebukuro,
Electrochemistry
2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 348, 352 - Electrochemical Detection of Vascular Endothelial Growth Factor with Aptamer Sandwich
Yoshihiko Nonaka, Koichi Abe and Kazunori Ikebukuro,
Electrochemistry
2012年05月, 研究論文（学術雑誌）, 共同, 80, 5, 363, 366 - BioRadioTransmitter: a self-powered wireless glucose-sensing system
Takuya Hanash, Tomohiko Yamazaki, Wakako Tsugawa, Kazunori Ikebukuro, Koji Sode
J. Diabetes Sci. Technol.
2011年09月, 研究論文（学術雑誌）, 共同, 5, 5, 1030, 1035 - Control of aptamer function using radiofrequency magnetic field,
Taira K, Abe K, Ishibasi T, Sato K, Ikebukuro K.,
J Nucleic Acids. 2011;2011:
2011年09月, 研究論文（学術雑誌）, 共同, 2011, 103872. - Development of a novel biosensing system based on the structural change of a polymerized guanine-quadruplex DNA nanostructure,
Morita Y, Yoshida W, Savory N, Han SW, Tera M, Nagasawa K, Nakamura C, Sode K, Kazunori Ikebukuro (2011)
Biosens. Bioelectron., 26(12)
2011年08月, 研究論文（学術雑誌）, 共同, 26, 12, 4837, 41 - Analysis of the unbinding force between telomestatin derivatives and human telomeric G-quadruplex by atomic force microscopy,
Amemiya Y, Furunaga Y, Iida K, Tera M, Nagasawa K, Ikebukuro K, Nakamura C.
Chem Commun (Camb)
2011年07月, 研究論文（学術雑誌）, 共同, 47, 26, 7485, 7 - Aptameric sensors based on structural change for diagnosis
Koichi Abe, Daisuke Ogasawara, Wataru Yoshida, Koji Sode, Kazunori Ikebukuro
Faraday Discuss.
2011年02月, 研究論文（学術雑誌）, 共同, 149, 93, 105 - A Non-label homogeneous protein detection based on laser interferometric photo-thermal displacement measurement using aptamers,
Kaori Tsukakoshi, Daisuke Ogasawara, Eiji Takahashi, Ryo Katayama, Kazunori Ikebukuro
Biotechnology Journal,10
2011年01月, 研究論文（学術雑誌）, 共同, 6, 1, 101, 116. - Selection of DNA aptamer against prostate specific antigen using a genetic algorithm and application to sensing,
Nasa Savory, Koichi Abe, Koji Sode, Kazunori Ikebukuro
Biosens. Bioelectron.
2010年05月, 研究論文（学術雑誌）, 共同, 26, 4, 1386, 1391. - Constructing an improved pyrroloquinoline quinone glucose dehydrogenase binding aptamer for enzyme labeling
Koichi Abe, Koji Sode, Kazunori Ikebukuro
Biotechnol. Lett.
2010年05月, 研究論文（学術雑誌）, 共同, 32, 9, 1293, 1298. - Visualization of G-quadruplexes by using a BODIPY-labeled macrocyclic heptaoxazole
Masayuki Tera, Iida K, Ikebukuro K, Seimiya H, Shin-Ya K, Nagasawa K
Org Biomol Chem,
2010年02月, 研究論文（学術雑誌）, 共同, 8, 12, 2749, 2755 - Screening of DNA aptamer which binds to alpha-synuclein
Kaori Tsukakoshi Ryuichi Harada, Koji Sode and Kazunori Ikebukuro
Biotechnol. Lett.
2010年02月, 研究論文（学術雑誌）, 共同, 32, 5, 643, 648. - Screening and improvement of an anti-VEGF DNA aptamer,
Yoshihiko Nonaka, Koji Sode and Kazunori Ikebukuro
Molecule
2010年02月, 研究論文（学術雑誌）, 共同, 15, 1, DOI(公開)(r-map), 215, 225 - Pyrroloquinoline quinone inhibits the fibrillation of amyloid proteins
3. Jihoon Kim, Masaki Kobayashi, Makoto Fukuda, Daisuke Ogasawara, Natsuki Kobayashi, Sungwoong Han, Chikashi Nakamura, Masaki Inada, Chisato Miyaura, Kazunori Ikebukuro and Koji Sode
Prion
2010年02月, 研究論文（学術雑誌）, 共同, 4, 1, DOI(公開)(r-map), Epub ahead of print - The effect of amino acid substitution in the imperfect repeat sequences of alpha-synuclein on fibrillation
Ryuichi Haradaa, Natsuki Kobayashia, Jihoon Kima, Chikashi Nakamurab, Sung-Woong Hanb, Kazunori Ikebukuroa and Koji Sode (2009)
Biochim Biophys Acta
2009年10月, 研究論文（学術雑誌）, 共同, 1792, 10, DOI(公開)(r-map), 998, 1003. - Kinetic mechanism and inhibitor characterization of WNK1 kinase
Yukiko I. Yagi*, Koichi Abe, Kazunori Ikebukuro and Koji Sode
Biochemistry
2009年10月, 研究論文（学術雑誌）, 共同, 48, 43, DOI(公開)(r-map), 10255, 10266. - Zn finger-based direct detection system for PCR products of Salmonella spp. and the influenza A virus
Yuko Osawa, Hiroaki Motoki, Takafumi Matsuo, Michio Horiuchi, Koji Sode, Kazunori Ikebukuro
Biotechnolgy Leterrs
2009年02月, 研究論文（学術雑誌）, 共同, 31, 5, 725, 733 - An Aptamer-Based Bound/Free Separation System for Protein Detection, Electroanalysis
Mieko Fukasawa, Wataru Yoshida, Hiroki Yamazaki, Koji Sode, and Kazunori Ikebukuro,
Electroanalysis
2009年01月, 研究論文（学術雑誌）, 共同, 21, 11, DOI(公開)(r-map), 1297, 1302. - DNA aptamers that bind to PQQGDH as an electrochemical labeling tool
Yuko Osawa, Madoka Takase, Koji Sode, Kazunori Ikebukuro
Electroanalysis
2009年01月, 研究論文（学術雑誌）, 共同, 21(11),, 11, DOI(公開)(r-map), 1303, 1308. - BioCapacitor-A novel category of biosensor.
Hanashi T, Yamazaki T, Tsugawa W, Ferri S, Nakayama D, Tomiyama M, Ikebukuro K, Sode K
Biosensors & Bioelectronics
2008年11月, 研究論文（学術雑誌）, 共同, E pub ahead of print, DOI(公開)(r-map) - コンピューター内進化を用いたアプタマーの改良
池袋一典
日本化学会情報化学部会誌「CICSJ Bulletin」
2008年10月, 単独, 26, 3, 5, 8 - Zinc finger protein-based detection system of PCR products for pathogen diagnosis
Yuko Osawa, Hiroaki Motoki, Takafumi Matsuo, Michio Horiuchi, Koji Sode, Kazunori Ikebukuro
Nucleic Acids Symp Ser (Oxf
2008年09月, 研究論文（学術雑誌）, 共同, 2008, 52, 23, 24 - Detection system based on the conformational change in an aptamer and its application to simple bound/free separation
Ogasawara D, Hachiya NS, Kaneko K, Sode K, Ikebukuro K
Biosensors & Bioelectronics
2008年09月, 研究論文（学術雑誌）, 共同, E pub ahead of print, DOI(公開)(r-map) - Aggregation and fibrillation study of alpha-synuclein under applied voltage
Osawa, Yuko, Ikebukuro, Kazunori, Kobayashi, Natsuki, Han, SungWoong, Nakamura, Chikashi, Sode, Koji
ELECTROCHEMISTRY
2008年08月, 研究論文（学術雑誌）, 共同, 76, 8, 614, 618 - Selection of DNA aptamers against insulin and construction of an aptameric enzyme subunit for insulin sensing
Wataru Yoshida, Eriko Mochizuki, Hiroki, Yamazaki, Koji Sode, Kazunori Ikebukuro
Biosensors & Bioelectronics
2008年06月, 研究論文（学術雑誌）, 共同, E pub ahead of print, DOI(公開)(r-map) - The simple and rapid detection of specific PCR products from bacterial genomes using Zn finger proteins
Yuko Osawa, Kazunori Ikebukuro, Hiroaki Motoki, Takafumi Matsuo, Michio Horiuchi, Koji Sode
Nucleic acids research
2008年04月, 研究論文（学術雑誌）, 共同, 36, 11, e68 - Improvement of aptamer affinity by dimerization
Hijiri Hasegawa, Ken-Ichi Taira, Koji Sode, Kazunori Ikebukuro
Sensors
2008年02月, 研究論文（学術雑誌）, 共同, 8, 5, 1090, 1098 - Selection of DNA aptamers against VEGF(165) using a protein competitor and the aptamer blotting method
Hijiri Hasegawa, Koji Sode, Kazunori Ikebukuro
Biotechnology Letters
2008年01月, 研究論文（学術雑誌）, 共同, 30, 5, 829, 834 - Screening of DNA aptamer against mouse prion protein by competitive selection
Daisuke Ogasawara, Hijiri Hasegawa, Kiyotoshi Kaneko, Koji Sode, and Kazunori Ikebukuro
Prion
2007年12月, 研究論文（学術雑誌）, 共同, 1, 4, 248, 254 - Selection of DNA aptamers that inhibit enzymatic activity of PQQGDH and its application
Kazunori Ikebukuro, Madoka Takase, and Koji Sode
Nucleic Acids Symp Ser,
2007年11月, 研究論文（学術雑誌）, 共同, 51:, 403, 404. - Construction of target molecule sensing system using aptameric enzyme subunit based on PQQGDH activity
Kazunori Ikebukuro, Yo Morita, Wataru Yoshida, and Koji Sode
Nucleic Acids Symp Ser,
2007年11月, 研究論文（学術雑誌）, 共同, 51:, 401, 402. - Selection and characterization of DNA aptamers against VEGF165 with aptamer blotting method and its application
Kazunori Ikebukuro, Hijiri Hasegawa, and Koji Sode
Nucleic Acids Symp Ser,
2007年11月, 研究論文（学術雑誌）, 共同, 51:, 399, 400. - Specific detection of PCR product from Legionella pneumophila strainPhiladelphia1 using zinc finger protein Sp2
Kazunori Ikebukuro, Takenori Kumagai, Hiroaki Motoki, Yuko Osawa, Takafumi Matuo, Michio Horiuchi, and Koji Sode
Nucleic Acids Symp Ser,
2007年11月, 研究論文（学術雑誌）, 共同, 51:, 285, 286 - Aptameric enzyme subunit for homogeneous protein sensing
Wataru Yoshida, Koji Sode, and Kazunori Ikebukuro
Nucleic Acids Symp Ser
2007年11月, 研究論文（学術雑誌）, 共同, 51:, 99, 100. - Peptide ligand screening of alpha-synuclein aggregation modulators by in silico panning
Koichi Abe, Natsuki Kobayashi, Koji Sode and Kazunori Ikebukuro
BMC bioinformatics
2007年11月, 研究論文（学術雑誌）, 共同, 8, 45 - Label free homogeneous detection of IgE by aptameric enzyme subunit
Wataru Yoshida, Kazunori Ikebukuro and Koji Sode
Biotechnology Letters
2007年11月, 研究論文（学術雑誌）, 共同, 30, 3, 829, 834 - Aptameric enzyme subunit for homogenious DNA sensing
Kazunori Ikebukuro, Wataru Yoshida and Koji Sode
Biotechnology Letters
2007年10月, 研究論文（学術雑誌）, 共同, 30, 2, 243, 252 - Stopped-flow system with ozonizer for the estimation of low biochemical oxygen demand in environmental samples
Chee GJ, Nomura Y, Ikebukuro K, Karube I.
Biosens Bioelectron.
2007年01月, 研究論文（学術雑誌）, 共同, 22, 2, 3092, 3098 - In silico panning for a non-competitive peptide inhibitor
Yukiko Yagi , Kotaro Terada, Takahisa Noma , Kazunori Ikebukuro and Koji Sode
BMC Bioinformatics
2007年01月, 研究論文（学術雑誌）, 共同, 8, 1, 11 - Pyrroloquinoline quinone (PQQ) prevents fibril formation of alpha-synuclein
Biochemical Biophysical Research Communication
2006年10月, 研究論文（学術雑誌）, 共同, 349, 3, 1139, 1144 - Analysis of DNA and zinc finger interactions using mechanical force spectroscopy
Yanyan Wang, Shin-ichiro Oyokawa, SungWoong Han, Wei Huang, Kazunori Ikebukuro, Chikashi Nakamura and Jun Miyake
NanoBiotechnology
2006年09月, 研究論文（学術雑誌）, 共同, 2, 3-4, 87, 94 - Homogeneous DNA sensing using enzyme-inhibiting DNA aptamers
1. Wataru Yoshida, Koji Sode and Kazunori Ikebukuro
Biochemical and Biophysical Research Communications
2006年09月, 研究論文（学術雑誌）, 共同, 348, 1, 245, 252 - Analysis of the evolution of the thrombin-inhibiting DNA aptamers using a genetic algorithm
Kazunori Ikebukuro, Wataru Yoshida, Takahisa Noma and Koji Sode
Biotechnology Letters
2006年09月, 研究論文（学術雑誌）, 共同, 28, 23, 1933, 1937 - Characterization and application of aptamers for Taq DNA polymerase selected using an evolution-mimicking algorithm
Takahisa Noma, Koji Sode
Biotechnology Letters
2006年09月, 研究論文（学術雑誌）, 共同, 28, 23, 1939, 1944 - Aptamer selection based on inhibitory activity using an evolution-mimicking algorithm
Takahisa Noma
Biochemical and Biophysical Research Communications
2006年07月, 研究論文（学術雑誌）, 共同, 347, 1, 226, 231 - A screening method of DNA aptamers that bind to a specific protein in tissue,
Takahisa Noma, Kazunori Ikebukuro, Koji Sode, Takuya Ohkubo, Yuji Sakasegawa, Naomi Hachiya and Kiyotoshi Kaneko
Biotechnology Letters
2006年05月, 研究論文（学術雑誌）, 共同, 28, 17, 1377, 1381 - Aptameric enzyme subunit for biosensing based on enzymatic activity measurement,
Wataru Yoshida, Koji Sode and Kazunori Ikebukuro
Analytical Chemistry
2006年05月, 研究論文（学術雑誌）, 共同, 78, 10, 3296, 3303 - Development of a novel sensing probe using DNA aptamer inhibiting enzymatic activity
Kazunori Ikebukuro, Wataru Yoshida, and Koji Sode
Nucleic Acids Symp Ser (Oxf)
2005年11月, 研究論文（学術雑誌）, 共同, 2005, 49, 83, 83 - Screening of DNA aptamers against multiple proteins in tissue
14. Takahisa Noma, Kazunori Ikebukuro, Koji Sode, Takuya Ohkubo, Yuji Sakasegawa, Naomi Hachiya and Kiyotoshi Kaneko
Nucleic Acids Symp Ser (Oxf)
2005年11月, 研究論文（学術雑誌）, 共同, 2005, 49, 357, 358 - A novel method of screening thrombin-inhibiting DNA aptamers using an evolution-mimicking algorithm
Kazunori Ikebukuro, Yuji Okumura, Koichi Sumikura and Isao Karube
Nucleic Acids Research
2005年08月, 研究論文（学術雑誌）, 共同, 33, 12, e108 - Development of photocatalytic biosensor for the evaluation of biochemical oxygen demand. ., 21(1), .
Gab-Joo Chee, Yoko Nomura, Kazunori Ikebukuro and Isao Karube
Biosens Bioelectron
2005年06月, 研究論文（学術雑誌）, 共同, 21, 7, 67, 73 - Development of a novel glucose enzyme fuel cell system employing protein engineered PQQ glucose dehydrogenase. .
Noriko Yuhashi, Masamitsu Tomiyama, Junko Okuda, Satoshi Igarashi, Kazunori Ikebukuro, and Sode, Koji
Biosens Bioelectron
2005年05月, 研究論文（学術雑誌）, 共同, 20, 10, 2145, 50 - Novel electrochemical sensor system for protein using the aptamers in sandwich manner
Kazunori Ikebukuro, Chiharu Kiyohara and Koji Sode
Biosens Bioelectron
2005年02月, 研究論文（学術雑誌）, 共同, 20, 10, 2168, 2172 - アプタマーを分子認識素子とするバイオセンサの開発」
池袋一典
次世代センサ14号2巻、p5-8、 (2005)
2005年01月, 研究論文（学術雑誌）, 単独, 14, 2, 5, 8 - Electrochemical Detection of Protein Using a Double Aptamer Sandwich
Kazunori Ikebukuro, Chiharu Kiyohara and Koji Sode
Analytical Letters
2004年12月, 研究論文（学術雑誌）, 共同, 37, 14, 2901, 2909 - Selection of DNA aptamers that inhibit Taq DNA polymerase by an algorithm mimicking evolution.
Takahisa Noma, Kazunori Ikebukuro and Koji Sode
Nippon Kessho Seicho Gakkaishi
2004年12月, 研究論文（学術雑誌）, 共同, 31, 3, 190 - Development of Salmonella gyrB PCR product detection method using Zn finger protein
Tomohiko Yabata, Kazunori Ikebukuro and Koji Sode
Nippon Kessho Seicho Gakkaishi
2004年12月, 研究論文（学術雑誌）, 共同, 31, 3, 189 - Development of a novel DNA sensing system using DNA aptamer inhibited enzymatic activity 1
Kazunori Ikebukuro, Wataru Yoshida and Koji Sode
Nucleic Acids Symp Ser (Oxf),
2004年11月, 研究論文（学術雑誌）, 共同, 2004, 48, 231, 232 - Development of a novel DNA sensing system using DNA aptamer that inhibits enzymatic activity 2
Kazunori Ikebukuro, Wataru Yoshida, Takahisa Noma and Koji Sode
Nucleic Acids Symp Ser (Oxf)
2004年11月, 研究論文（学術雑誌）, 共同, 2004, 48, 309, 310 - 英語学術論文 その他31報 合計103報 英語著書 合計４編
2004年10月, 単独 - Molecular engineering of PQQGDH and its applications
Satoshi Igarashi, Junko Okuda, Kazunori Ikebukuro and Koji Sode
Archives of Biochemistry and Biophysics
2004年09月, 研究論文（学術雑誌）, 共同, 428, 1, 52, 63 - Single nucleotide polymorphism typing on DNA array with hydrophobic surface fabricated by plasma-polymerization technique,
Hirotaka Miyachi, Kazunori Ikebukuro, Kazuyoshi Yano, Hiroyuki Aburatani and Isao Karube
Biosens. Bioelectron.
2004年07月, 研究論文（学術雑誌）, 共同, 20, 2, 184, 189 - Development of an enzymatic flow-injection chemiluminescence system for determining inorganic pyrophosphate ion,
Hideaki Nakamura, Ruriko Yamazaki, Takayuki Shirai, Hisae Sano, Yu Nakami, Kazunori Ikebukuro, Kazuyoshi Yano, Yoko Nomura, Yasushi Hasebe, Yoshiko Arikawa, Yuzo Masuda, and Isao Karube
ANALYTICA CHIMICA ACTA
2004年03月, 研究論文（学術雑誌）, 共同, 518, 45, 49. - PCR-Based Ribosomal DNA Detection Technique for Microalga (Heterosigma carterae) Causing Red Tide and Its Application to a Biosensor Using Labeled Probe,
Ryoichi Asai, Kozue Ootani, Yoko Nomura, Chikashi Nakamura, Kazunori Ikebukuro, , Yoshiko Arikawa, Jun Miyake and Isao Karube
Marine Biotechnology.
2003年11月, 研究論文（学術雑誌）, 共同, 5, 5, 417, 423 - Screening of DNA aptamers inhibiting Taq DNA polymerase using algorithm mimicking evolution,
Kazunori Ikebukuro and Noma, Takahisa
Nucleic Acids Research Supplement 3(3rd International Symposium on Nucleic Acids Chemistry [and] 30th Symposium on Nucleic Acids Chemistry in Japan, 2003),
2003年11月, 研究論文（学術雑誌）, 単独, 2003, 309, 310. - Surface plasmon resonance probe with a novel integrated reference sensor surface,
Takuo Akimoto, Kazunori Ikebukuro and Isao Karube
Biosensors & Bioelectronics, 18(12), .
2003年10月, 研究論文（学術雑誌）, 共同, 18, 12, 1447, 1453 - A polymerase chain reaction-based ribosomal DNA detection technique using a surface plasmon resonance detector for a red tide causing microalga, Alexandrium affine,
8. Ryoichi Asai, Keijoro Nakanishi, Chikashi Nakamura, Jun Miyake, Isao Karube
Phycological Research,
2003年10月, 研究論文（学術雑誌）, 共同, 51, 2, 118, 125 - Preparation of a whole genome phage library using fragmented Escherichia coli genome and its characterization of protein binding properties by surface plasmon resonance
Kazuyoshi Yano, Masato Shionoya, Tetsuya Yoshino, Shinichi Y. Sawata, Kazunori Ikebukuro and Isao Karube
Biosensors & Bioelectronics, 18(10), 1201-1207
2003年10月, 研究論文（学術雑誌）, 共同, 18, 10, 1201, 1207 - Improvement of a CL-FIA system using maltose phosphorylase for the detemination of phosphate-ion in fresh water
Analytical Letters,
2003年09月, 研究論文（学術雑誌）, 共同, 36, 9, 1805, 1817 - A bioassay to detect contaminant-induced messenger RNA using a transcriptomic approach: Detection of RT-PCR-amplified single-stranded DNA based on the SPR sensor in cyanobacteria
Analytical Letters, 36(8), 1475-1491.
2003年08月, 研究論文（学術雑誌）, 共同, 36, 8, 1475, 1491 - (2003) Novel strategy for DNA aptamers inhibiting enzymatic activity using algorithm mimicking evolution,
Kazunori Ikebukuro, Yuji Okumura, Koichi Sumikura and Isao Karube
Nucleic Acids Research Supplement 3(3rd International Symposium on Nucleic Acids Chemistry [and] 30th Symposium on Nucleic Acids Chemistry in Japan, 2003)
2003年07月, 研究論文（学術雑誌）, 共同, 2003, 205, 206 - Amperometric glucose sensor using thermostable co-factor binding glucose dehydrogenase
Yukie Nakazawa, Tomohiko Yamazaki, Wakako Tsugawa, Kazunori Ikebukuro and Koji Sode
IEEJ Transactions on Sensors and Micromachinines, 123(6), 185-189
2003年06月, 共同 - Electrochemical DNA sensor using genetically engineered thermostable pyrroloquinoline quinone glucose dehydrogenase
Kazunori Ikebukuro, Yoko Saito, Satoshi Igarashi and Koji Sode
Electrochemistry, 71(6), 490-495.
2003年06月, 共同 - Molecular engineering of PQQGDH and its applications,
Satoshi Igarashi, Junko Okuda, Kazunori Ikebukuro and Koji Sode
Arch Biochem Biophys
2003年05月, 研究論文（学術雑誌）, 共同, 428, 1, 52, 63 - Electrochemical protein chip with arrayed immunosensors with antibodies immobilized in a plasma-polymerized film
Kojima K, Hiratsuka A, Suzuki H, Yano K, Ikebukuro K, Karube
Analytical Chemistry
2003年05月, 研究論文（学術雑誌）, 共同, 75, 5, 1116, 1122 - Development of a reactor type bio-sensor for trichloroethylene
Tae-Sung Han, Satoshi Sasaki, Kazuyoshi Yano, Kazunori Ikebukuro, Kitayama Atsushi, Teruyuki Nagamune and Isao Karube
Analytical Letters 36(3), 539-547.
2003年02月, 共同 - Exploration of structural features of monomeric helical peptides designed with a genetic algorithm
9. Wuming. Zhang, Michael. G. Loughran, Shu-ichi. Kanna, Kazuyoshi. Yano, Kazunori. Ikebukuro, Yohei Yokobayashi, Ryoko Kuroda and Isao Karube
Proteins: Structure, Function, and Genetics, 53(2), 193-200.
2003年01月, 研究論文（学術雑誌）, 共同, 53, 2, 193, 200. - コンピューター内進化を利用した酵素阻害DNAアプタマーの探索
化学と教育、51号1巻、p26-27
2003年01月, 単独 - Improved substrate specificity of water-soluble pyrroloquinoline quinone glucose dehydrogenase by a peptide ligand
Hiromi Yoshida, Yukiko Yagi, Kazunori Ikebukuro and Koji Sode
Biotechnology Letters, 25(4), 301-305.
2003年01月, 共同 - Flow injection microbial trichloroethylene sensor
Tae-Song Han, Satoshi Sasaki, Kazuyoshi Yano, Kazunori Ikebukuro, Atsushi Kitayama, Teruyuki Nagamune, Isao Karube
Talanta, 57(2), 271-276.
2002年12月, 共同 - A simple nitrate sensor system using titanium trichloride and an ammonium electrode
Seung-Jin Cho, Satoshi Sasaki, Kazunori Ikebukuro, Isao Karube
Sensors and Actuators, B: Chemical, 85(1-2), 120-125.
2002年08月, 共同 - A flow method with photocatalytic oxidation of dissolved organic matter using a solid-phase (TiO2) reactor followed by amperometric detection of consumed oxygen
Yoon-Chang Kim, Satoshi Sasaki, Kazuyoshi Yano, Kazunori Ikebukuro, Kazuhiko Hashimoto, Isao Karube
Analytical Chemistry, 74(15), 3858-3864.
2002年02月, 研究論文（学術雑誌）, 共同 - Detection technique of asymmetric RT-PCR-based amplified single-stranded DNA and its application to biosensor for detection of mRNA for cyanobacteria, Anabaena variabilis
Roichi Asai, Chikashi Nakamura, Kazunori Ikebukuro, Isao Karube and Jun Miyake
Biotechnology Letters, 24(20), 1677-1682.
2002年02月, 共同 - Amperometric DNA sensor using the pyrroquinoline quinone glucose dehydrogenase ? avidin conjugate
Kazunori Ikebukuro, Yumiko Kohiki and Koji Sode
Biosensors and Bioelectronics
2002年01月, 共同, 17, 11-12, 1075, 1080 - Detection of phycobilin pigments and their seasonal change in Lake Kasumigaura using a sensitive in situ fluorometric sensor
Asai R, Horiguchi Y, Yoshida A, McNiven S, Tahira P, Ikebukuro K, Uchiyama S, Masuda Y, Karube I
Analytical Letters
2001年11月, 研究論文（学術雑誌）, 共同, 34, 14, 2521, 2533 - Dioxin detection based on immunoassay using a polyclonal antibody against octa-chloricated dibenzo-p-dioxin (OCDD)
Mifumi shimomura, Yoko Nomura, Kyong-Hoon Lee, Kazunori Ikebukuro and Isao Karube
Analyst,
2001年11月, 共同, 126, 1207, 1209. - Microbial sensor for trichloroethylene determination
Tae-Song Han, Yoon-Chang Kim, Satoshi Sasaki, Kazuyoshi Yano, Kazunori Ikebukuro, Atsushi Kitayama, Teruyuki Nagamune, Isao Karube
Analytica Chimica Acta, 431(2), 225-230.
2001年08月, 共同, 431, 2, 225, 230 - Simple and rapid detection method using surface plasmon resonance for dioxins, polycholorinated biphenylx and atrazine
Mifumi shimomura, Yoko Nomura, Wei Zhang, Masato Sakino, Kyong-Hoon Lee, Kazunori Ikebukuro and Isao Karube
Analytica Chimica Acta,
2001年07月, 共同, 434, 223, 230 - A fluorescent nitrate sensing system using a reaction cartridge and titanium trichloride
Seung-Jin Cho, Satoshi Sasaki, Kazunori Ikebukuro, Isao Karube
Talanta,
2001年06月, 共同, 54, 5, 903, 911. - Photocatalytic sensor for the determination of chemical oxygen demand using flow injection analysis
Yoon-Chang Kim, Satoshi Sasaki, Kazuyoshi Yano, Kazunori Ikebukuro, Kazuhiko Hashimoto, Isao Karube
Analytica Chimica Acta,
2001年05月, 共同, 432, 1, 59, 66 - Biosensor for the evaluation of biochemical oxygen demand using photocatalytic pretreatment
Gab-Joo Chee, Yoko Nomura, Kazunori Ikebukuro and Isao Karube
Sensors and Actuators, B: Chemical,
2001年04月, 共同, 80,, 15, 20. - Integration of microfabricated needle-type glucose sensor devices with a novel thin-film Ag/AgCl electrode and plasma-polymerized thin film: mass production techniques.
Atsunori Hiratsuka, Kenichi Kojima, Hiroaki Suzuki, Hitoshi Muguruma, Kazunori Ikebukuro and Isao Karube
The Analyst
2001年03月, 共同, 126, 5, 658, 663 - Detection of toxic chemicals with high sensitivity by measuring the quantity of induced P450 mRNAs based on surface plasmon resonance.
Masaaki Oyama, Takeshi Ikeda, Tae-Kyu Lim, Kazunori Ikebukuro, Yuzo Masuda and Isao Karube
Biotechnol. Bioeng
2001年01月, 共同, 71, 3, 217, 222 - Application of chimeric RNA-DNA oligonucleotides to the detection of pathogenic microorganisms using surface plasmon resonance
Hirotaka Miyachi, Kazuyoshi Yano, Midori Kono, Sadayori Hoshina, Kazunori Ikebukuro and Isao Karube
Analytica Chimica Acta, 407 (1-2), 1-10.
2000年, 共同, 407, 1-2, 1, 10 - Highly sensitive detection of atrazine based on the SPR determination of P450 mRNA levels in Saccharomyces cerevisiae.
Tae-Kyu Lim, Masaaki Oyama, Kazunori Ikebukuro and Isao Karube
Analytical Chemistry, 72 (13), 2856-2860.
2000年, 共同 - Relationship between theoretical oxygen demand and photocatalytic chemical oxygen demand for specific classes of organic chemicals.
Yoon-Chang Kim, Satoshi Sasaki, Kazuyoshi Yano, Kazunori Ikebukuro, Kazuhiko Hashimoto and Isao Karube
The Analyst, 125 (11), 1915-1918.
2000年10月, 共同 - Disposable type chemical oxygen demand sensor using a microfabricated Clark-type oxygen electrode with a TiO2 suspension solution.
Kyong-Hoon Lee, Yoon-Chang Kim, Hiroaki Suzuki, Satoshi Sasaki, Kazunori Ikebukuro, Kazuhiko Hashimoto and Isao Karube
Electroanalysis, 12 (16), 1334-1338.
2000年09月, 共同 - Development of a fluorometric sensor for the measurement of phycobilin pigment and application to freshwater phytoplankton.
Ryoichi Asai, Scott McNiven, Kazunori Ikebukuro, Isao Karube, Yasuo Horiguchi, Shuichi Uchiyama, Akira Yoshida and Yuzo Masuda
Field Analytical Chemistry and Technology, 4 (1), 53-61.
2000年08月, 共同 - Effect of incident angle of a light on sensitivity and detection limit for antibody layer with surface plasmon resonance spectroscopy
Takuo Akimoto, Satoshi Sasaki, Kazunori Ikebukuro and Isao Karube
Biosensors & Bioelectronics, 15 (7-8), 355-362.
2000年07月, 共同 - Estimation of sensitivity for refractive index and immunoreaction in a surface plasmon resonance sensor probe
Takuo Akimoto, Satoshi Sasaki, Kazunori Ikebukuro and Isao Karube
Analytica Chimica Acta, 417 (2), 125-131.
2000年06月, 共同 - Photocatalytic sensor for chemical oxygen demand determination based on oxygen electrode.
Yoon-Chang Kim, Kyong-Hoon Lee, Satoshi Sasaki, Kazuhiko Hashimoto, Kazunori Ikebukuro and Isao Karube
Analytical Chemistry, 72 (14), 3379-3382.
2000年05月, 共同 - Optical fiber biosensor for the determination of low biochemical oxygen demand.
Gab-Joo Chee, Yoko Nomura, Kazunori Ikebukuro and Isao Karube
Biosensors & Bioelectronics, 15 (7-8), 371-376.
2000年05月, 共同 - Application of polymer-embedded proteins to fabrication of DNA array.
Hirotaka Miyachi, Atsunori Hiratsuka, Kazunori Ikebukuro, Kazuyoshi Yano, Hitoshi Muguruma andIsao Karube
Biotechnology and Bioengineering. 69 (3),323-329.
2000年03月, 共同 - Bioassay of bile acids using an enzyme-linked DNA aptamer.
Teru Kato, Kazuyoshi Yano, Kazunori Ikebukuro and Isao Karube
The Analyst, 125, 1371-1373.
2000年02月, 共同 - Increasing the sensitivity of piezoelectric odor sensors based on molecularly imprinted polymers.
Hong-Seok Ji, Scott McNiven, Takashi Saito, Kazunori Ikebukuro, Isao Karube
Biosensors & Bioelectronics, 15 (7-8), 403-409.
2000年02月, 共同 - Detection of PCR products of Escherichia coli O157:H7 in human stool samples using surface plasmon resonance (PCR).
Eriko Kai, Kazunori Ikebukuro, Sadayori Hoshina, Haruo Watanabe and Isao Karube
FEMS Immunology and Medical Microbiology., 29 (4), 283-288.
2000年02月, 共同 - In vitro selection of DNA aptamers which bind to cholic acid.
Teru Kato, Taro Takemura, Kazuyoshi Yano, Kazunori Ikebukuro and Isao Karube
Biochimica et Biophysica Acta, gene structure and expression, 1493, 12-18.
2000年01月, 研究論文（学術雑誌）, 単独 - Interaction of three-way DNA junctions with steroids.
Teru Kato, Kazuyoshi Yano, Kazunori Ikebukuro and Isao Karube
Nucleic Acids Research, 28(9), 1963-1968.
2000年01月, 研究論文（学術雑誌）, 共同 - 遺伝的アルゴリズムによる機能性分子の探索
生物工学会誌、日本生物工学会、72巻、3号、p210、(1999)
1999年10月, 単独 - Highly sensitive chemiluminescence flow-injection detection of the red tide phytoplankton Heterosigma carterae.
Ryoichi Asai, Ritsuko Matsukawa, Kazunori Ikebukuro and Isao Karube
Analytica Chimica Acta, 390 (1-3), 237-244.
1999年08月, 共同 - Refractive-index and thickness sensitivity in surface plasmon resonance spectroscopy.
Takuo Akimoto, Satoshi Sasaki, Kazunori Ikebukuro and Isao Karube
Applied Optics, 38 (19), 4058-4064.
1999年07月, 共同 - Selective piezoelectric odor sensors using molecularly imprinted polymers.
Hong-Seok Ji, Scott McNiven, Kazunori Ikebukuro and Isao Karube
Analytica Chimica Acta, 390 (1-3), 93-100.
1999年05月, 共同 - Highly sensitive trilayer piezoelectric odor sensor.
Hong-Seok Ji, Scott McNiven, Kazuyoshi Yano, Kazunori Ikebukuro, Uwe T. Bornscheuer, Rolf D. Schmid and Isao Karube
Analytica Chimica Acta, 387 (1), 39-45.
1999年05月, 共同 - Detection of Escherichia coli O157 : H7 DNA using two fluorescence polarization methods.
Bang-Ce Ye, Kazunori Ikebukuro and Isao Karube
Fresenius J. Analytical Chemistry., 365(5) 452-457.
1999年04月, 共同 - Application of peptide nucleic acid to the direct detection of deoxyribonucleic acid amplified by the polymerase chain reaction.
Shinya Sawata, Eriko Kai, Kazunori Ikebukuro, Tetsuya Iida, Takeshi Honda and Isao Karube
Biosensors and Bioelectronics, 14 (4), 397-404.
1999年03月, 共同 - 1999) Development of highly sensitive BOD sensor and its evaluation using preozonation.
Gab-Joo Chee, Yoko Nomura, Kazunori Ikebukuro and Isao Karube
Analytica Chimica Acta, 394, 65-71.
1999年02月, 共同 - An automatic flow-injection analysis system for determining phosphate ion in river water using pyruvate oxidase G (from Aerococcus viridans).
Hideaki Nakamura, Hiroko Tanaka, Mami Hasegawa, Yuzo Masuda, Yoko Nomura, Yoshiko Arikawa, Ritsuko Matsukawa, Kazunori Ikebukuro and Isao Karube
Talanta, 50(4), 799-807.
1999年02月, 共同 - A novel method for detecting PCR products in solution using surface plasmon resonance.
Eriko Kai, Shinya Sawata, Kazunori Ikebukuro, Tetsuya Iida, Takeshi Honda and Isao Karube
Analytical Chemistry., 71 (4), 796-800.
1999年02月, 研究論文（学術雑誌）, 共同 - Development of a highly sensitive chemiluninescent flow-injection analysis sensor for phosphate ion detection using maltose phosphorylase.
Hideaki Nakamura, Mami Hasegawa, Yoko Nomura, Yoshiko Arikawa, Ritsuko Matsukawa, Kazunori Ikebukuro and Isao Karube
Journal of Biotechnology, 75(2-3), 127-133.
1999年01月, 共同 - (1999) A glucose sensor with a plasma-polymerized thin film by dry processes.
Atsunori Hratsuka, Hitoshi Muguruma, Kazunori Ikebukuro and Isao Karube
Electroanalysis, 11(15), 1098-1100.
1999年01月, 共同 - Chemiluminescence flow-injection system for rapid detection of red tide phytoplankton, Chattonella antiqua.
Ryoichi Asai, Ritsuko Matsukawa, Kazunori Ikebukuro and Isao Karube
Analytical Letters, 31 (13), 2279-2288.
1998年11月, 共同 - Direct measurement of etophenprox using surface plasmon resonance.
Satoshi Sasaki, Eriko Kai, Hirotaka Miyachi, Hitoshi Muguruma, Kazunori Ikebukuro, Hidero Ohkawa and Isao Karube
Analytica Chimica Acta, 363(2-3), 229-233.
1998年11月, 共同 - Development of an odorant sensor using polymer-coated quartz crystals modified with unusual lipids.
Kazuyoshi Yano, Uwe T. Bornscheuer, Rolf D. Schmid, Hiroshi Yoshitake, Hong-Seok Ji, Kazunori Ikebukuro, Yuzo Masuda and Isao Karube
Biosensors & Bioelectronics, 13(3-4), 397-405.
1998年10月, 共同 - Quantitative analysis of polymerase chain reaction using anisotropy ratio and relative hydrodynamic volume of fluorescence polarization method.
Bang-Ce Ye Kazunori Ikebukuro and Isao Karube
Nucleic Acids Research, 26 (15), 3614-3615.
1998年10月, 研究論文（学術雑誌）, 共同 - Application of a linear alkylbenzene sulfonate biosensor to river water monitoring.
Yoko Nomura, Kazunori Ikebukuro, Kenji Yokoyama, Toshihumi Takeuchi, Yoko Arikawa, Sizue Ohno and Isao Karube.
Biosensors & Bioelectronics, 13, 1047-1053.
1998年09月, 共同 - Purification of single stranded DNA from asymmetric PCR product using the SMARTTM system.
Eriko Kai, Koichi Sumikura Kazunori Ikebukuro and Isao Karube
Biotechnology Techniques, 12 (12), 935-939.
1998年09月, 共同 - Cloning and expression of gene encoding cyanidase from Pseudomonas stutzeri AK61.
Atsushi Watanabe, Kazuyoshi Yano Kazunori Ikebukuro and Isao Karube
Applied Microbiology and Biotechnology, 50 (1), 93-97.
1998年08月, 共同 - Purification and characterization of a cyanide-degrading enzyme from Pseudomonas stutzeri AK61.
Atsushi Watanabe, Kazuyoshi Yano Kazunori Ikebukuro and Isao Karube
Microbiology, 144(6), 1677-1682.
1998年06月, 共同 - Investigation of the potential active site of a cyanide dihydratase using site-directed mutagenesis.
Atsushi Watanabe, Kazuyoshi Yano Kazunori Ikebukuro Isao Karube
Biochimica et Biophysica Acta, 1382(1), 1-4.
1998年05月, 研究論文（学術雑誌）, 共同 - Photogeneration of NADPH by oligothiophenes coupled with Ferredoxin-NADPH reductase.
Young-Sug Kim, Kazunori Ikebukuro, Hitoshi Muguruma and Isao Karube
Journal of Biotechnology, 59(3), 213-220.
1998年04月, 共同 - Molecularly imprinted polymers which mimic multiple hydrogen bonds between nucleotide bases.
Kazuyoshi Yano, Koichiro Tanabe, Toshifumi Takeuchi, Jun Matsui Kazunori Ikebukuro and Isao Karube
Analytica Chimica Acta, 363(2-3), 111-117.
1998年04月, 共同 - Selective recognition of 2,4 -dichlorophenoxyacetic acid using a molecularly imprinted polymer.
Yoko Nomura, Hitoshi Muguruma, Kazuyoshi Yano, Akimitsu Kugimiya, Scott McNiven and Kazunori Ikebukuro Isao Karube
Analytical Letters, 31(6), 973-980.
1998年03月, 共同 - Reagentless phosphate ion sensor system for environmental monitoring.
Suzuki Masayasu, Hironobu Kurata, Yoshihiro Inoue, Izumi Kubo, Hideaki Nakamura Kazunori Ikebukuro and Isao Karube
DENKI KAGAKU, 66(6) 579-583.
1998年02月, 研究論文（学術雑誌）, 共同 - Development of a liquid chromatography-based versatile bioanalysis for bevacizumab based on pretreatment combining aptamer affinity purification and centrifugal ultrafiltration concentration
Todoroki, Kenichiro; Hamada, Daichi; Yamada, Tomohiro; Saito, Taro; Shimizu, Yutaka; Sugiyama, Eiji; Mizuno, Hajime; Hayashi, Hideki; Tsukakoshi, Kaori; Ikebukuro, Kazunori
ANALYTICAL SCIENCES
SPRINGERNATURE
We report on the development of a versatile and accurate bioanalytical method for bevacizumab using a pretreatment method combining affinity purification with anti-idiotypic DNA aptamers and centrifugal ultrafiltration concentration, followed by liquid chromatography (LC)-fluorescence analysis. An affinity purification method using Sepharose beads as an affinity support removed immunoglobulin G and a large amount of coexisting substances in the serum sample. Purified bevacizumab was separated as a single peak by conventional LC and detected fluorometrically, showing good linearity (R2 = 0.999) in the range of 5-200 & mu;g/mL, sufficient to analyze bevacizumab concentrations in the blood of bevacizumab-treated patients. By combining this purification method with a concentration method using a centrifugal filtration device that inhibits non-specific adsorption of bevacizumab, the quantitative range was reduced by a factor of 10 while showing good linearity (R2 = 0.999) in the 0.5-20 & mu;g/mL range. The developed analytical method is expected to be used not only for general bioanalysis of therapeutic mAbs in clinical settings, but also for next-generation antibody drugs that show drug efficacy at low concentrations and for analysis of trace samples.
2023年11月, 研究論文（学術雑誌）, 共同, 39, 11, 0910-6340, DOI(公開)(r-map), 1805, 1811 - Regulation of thrombin activity by ligand-induced topological alteration in a thrombin-binding aptamer
Sasaki, Shogo; Ma, Yue; Hirokawa, Takatsugu; Ikebukuro, Kazunori; Tera, Masayuki; Nagasawa, Kazuo
CHEMICAL COMMUNICATIONS
ROYAL SOC CHEMISTRY
Thrombin-binding aptamer (TBA), which forms a G-quadruplex (G4) structure with anti-parallel topology, interacts with thrombin to inhibit its enzymatic activity. Here we show that the G4-topology-altering ligand L2H2-2M2EA-6LCO (6LCO) changes the anti-parallel topology of TBA G4 to the parallel topology, thereby abrogating the thrombin-inhibitory activity of TBA. This finding suggests that G4 ligands that alter topology may be promising drug candidates for diseases involving G4-binding proteins.
2023年07月13日, 研究論文（学術雑誌）, 共同, 59, 57, 1359-7345, DOI(公開)(r-map), 8862, 8865 - Protein engineering of antibody fragments for pharmaceutical production
Kuwahara, Atsushi; Ikebukuro, Kazunori; Asano, Ryutaro
APPLIED PHYSICS REVIEWS
AIP Publishing
Antibody fragments without the Fc region are attracting attention in the pharmaceutical industry due to their high ability to penetrate solid tissues, cost-effective expression using microbial expression systems, and distinctive modes of action compared to those of full-size antibodies. Based on these characteristics, several antibody fragment agents have been approved. However, developing platform engineering methodologies to accelerate their development is important. In this review, we summarize and discuss protein engineering strategies for preparing therapeutic antibody fragments composed of antibody variable domains. Three (introduction of high-solubility tag systems, complementarity-determining region grafting, and domain arrangements) and two (introduction of purification tag systems and mutagenesis studies for protein L- or protein A-binding) protein engineering strategies have been reported for the cultivation and purification processes, respectively. Fusion tags might negatively impact molecular folding, function, immunogenicity, and final yield. If the production behavior of antibody fragments is not improved through complementarity-determining region grafting, domain arrangements, or human sequence-based mutagenesis, using additional fusion tag systems should be considered, with careful attention to the points described above. This summarized knowledge regarding protein engineering strategies for effectively producing antibody fragments will further accelerate therapeutic antibody fragment development.
2023年09月, 研究論文（学術雑誌）, 共同, 10, 3, 1931-9401, DOI(公開)(r-map) - Real-time monitoring of the amyloid β1-42 monomer-to-oligomer channel transition using a lipid bilayer system
Numaguchi, Yuri; Tsukakoshi, Kaori; Takeuchi, Nanami; Suzuki, Yuki; Ikebukuro, Kazunori; Kawano, Ryuji; Wand, Josh
PNAS NEXUS
OXFORD UNIV PRESS
This study describes the observation of the transformation of monomeric amyloid β1–42 (Aβ42) into oligomers in a lipid membrane utilizing a lipid bilayer system for electrophysiological measurement. The relevance of oligomers and protofibrils in Alzheimer's disease (AD) is underscored given their significant neurotoxicity. By closely monitoring the shift of Aβ42 from its monomeric state to forming oligomeric channels in phospholipid membranes, we noted that this transformation transpired within a 2-h frame. We manipulated the lipid membrane's constitution with components such as glycerophospholipid, porcine brain total lipid extract, sphingomyelin (SM), and cholesterol (Chol.) to effectively imitate nerve cell membranes. Interesting findings showcased Chol.'s ability to foster stable oligomeric channel formation in the lipid membrane, with SM and GM1 lipids potentially enhancing channel formation as well. Additionally, the study identified the potential of a catechin derivative, epigallocatechin gallate (EGCG), in obstructing oligomerization. With EGCG present in the outer solution of the Aβ42-infused membrane, a noteworthy reduction in channel current was observed, suggesting the successful inhibition of oligomerization. This conclusion held true in both, prior and subsequent, stages of oligomerization. Our findings shed light on the toxicity of oligomers, promising invaluable information for future advancements in AD treatment strategies.
2023年12月14日, 研究論文（学術雑誌）, 共同, 3, 1, DOI(公開)(r-map), 13066, 13069 - Identification of novel amyloidosis in dogs: α-S1-casein acquires amyloidogenicity in mammary tumor by overexpression and N-terminal truncation
Murakami, Tomoaki; Kaku, Toshisuke; Tsukakoshi, Kaori; Iwaide, Susumu; Itoh, Yoshiyuki; Hisada, Miki; Nomura, Kohji; Kubo, Rikako; Ikebukuro, Kazunori; Sassa-O'Brien, Yukiko; Kametani, Fuyuki
VETERINARY PATHOLOGY
SAGE PUBLICATIONS INC
Mammary tumor-associated amyloidosis (MTAA) in dogs is characterized by amyloid deposition in the stroma of mammary adenoma or carcinoma; however, the amyloid precursor protein remains unknown. We attempted to identify an amyloid precursor protein and elucidated its etiology by characterizing 5 cases of canine MTAA. Proteomic analyses of amyloid extracts from formalin-fixed paraffin-embedded specimens revealed alpha-S1-casein (CASA1) as a prime candidate and showed the N-terminal truncation of canine CASA1. Both immunohistochemistry and immunoelectron microscopy showed that amyloid deposits or fibrils in MTAA cases were positive for CASA1. Reverse transcription-polymerase chain reaction and quantitative polymerase chain reaction revealed the complete mRNA sequence encoding CASA1, whose expression was significantly higher in the amyloid-positive group. The recombinant protein of the N-terminal-truncated canine CASA1 and the synthetic peptides derived from canine and human CASA1 formed amyloid-like fibrils in vitro. Structural prediction suggested that the N-terminal region of CASA1 was disordered. Previously, full-length CASA1 was reported to inhibit the amyloidogenesis of other proteins; however, we demonstrated that CASA1 acquires amyloidogenicity via excessive synthesis followed by truncation of its disordered N-terminal region. By identifying a novel in vivo amyloidogenic protein in animals and revealing key mechanistic details of its associated pathology, this study provides valuable insights into the integrated understanding of related proteopathies.
2023年03月, 研究論文（学術雑誌）, 共同, 60, 2, 0300-9858, DOI(公開)(r-map), 203, 213 - Lipopolysaccharide structure modulates cationic biocide susceptibility and crystalline biofilm formation in Proteus mirabilis
Clarke, O. E.; Pelling, H.; Bennett, V.; Matsumoto, T.; Gregory, G. E.; Nzakizwanayo, J.; Slate, A. J.; Preston, A.; Laabei, M.; Bock, L. J.; Wand, M. E.; Ikebukuro, K.; Gebhard, S.; Sutton, J. M.; Jones, B. V.
FRONTIERS IN MICROBIOLOGY
FRONTIERS MEDIA SA
Chlorhexidine (CHD) is a cationic biocide used ubiquitously in healthcare settings. Proteus mirabilis, an important pathogen of the catheterized urinary tract, and isolates of this species are often described as resistant to CHD-containing products used for catheter infection control. To identify the mechanisms underlying reduced CHD susceptibility in P. mirabilis, we subjected the CHD tolerant clinical isolate RS47 to random transposon mutagenesis and screened for mutants with reduced CHD minimum inhibitory concentrations (MICs). One mutant recovered from these screens (designated RS47-2) exhibited similar to 8-fold reduction in CHD MIC. Complete genome sequencing of RS47-2 showed a single mini-Tn5 insert in the waaC gene involved in lipopolysaccharide (LPS) inner core biosynthesis. Phenotypic screening of RS47-2 revealed a significant increase in cell surface hydrophobicity and serum susceptibility compared to the wildtype, and confirmed defects in LPS production congruent with waaC inactivation. Disruption of waaC was also associated with increased susceptibility to a range of other cationic biocides but did not affect susceptibility to antibiotics tested. Complementation studies showed that repression of smvA efflux activity in RS47-2 further increased susceptibility to CHD and other cationic biocides, reducing CHD MICs to values comparable with the most CHD susceptible isolates characterized. The formation of crystalline biofilms and blockage of urethral catheters was also significantly attenuated in RS47-2. Taken together, these data show that aspects of LPS structure and upregulation of the smvA efflux system function in synergy to modulate susceptibility to CHD and other cationic biocides, and that LPS structure is also an important factor in P. mirabilis crystalline biofilm formation.
2023年04月05日, 研究論文（学術雑誌）, 共同, 14, DOI(公開)(r-map) - Development of a Versatile Method to Construct Direct Electron Transfer-Type Enzyme Complexes Employing SpyCatcher/SpyTag System
Yanase, Takumi; Okuda-Shimazaki, Junko; Asano, Ryutaro; Ikebukuro, Kazunori; Sode, Koji; Tsugawa, Wakako
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
The electrochemical enzyme sensors based on direct electron transfer (DET)-type oxidoreductase-based enzymes are ideal for continuous and in vivo monitoring. However, the number and types of DET-type oxidoreductases are limited. The aim of this research is the development of a versatile method to create a DET-type oxidoreductase complex based on the SpyCatcher/SpyTag technique by preparing SpyCatcher-fused heme c and SpyTag-fused non-DET-type oxidoreductases, and by the in vitro formation of DET-type oxidoreductase complexes. A heme c containing an electron transfer protein derived from Rhizobium radiobacter (CYTc) was selected to prepare SpyCatcher-fused heme c. Three non-DET-type oxidoreductases were selected as candidates for the SpyTag-fused enzyme: fungi-derived flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase (GDH), an engineered FAD-dependent d-amino acid oxidase (DAAOx), and an engineered FMN-dependent l-lactate oxidase (LOx). CYTc-SpyCatcher (CYTc-SC) and SpyTag-Enzymes (ST-GDH, ST-DAAOx, ST-LOx) were prepared as soluble molecules while maintaining their redox properties and catalytic activities, respectively. CYTc-SC/ST-Enzyme complexes were formed by mixing CYTc-SpyCatcher and SpyTag-Enzymes, and the complexes retained their original enzymatic activity. Remarkably, the heme domain served as an electron acceptor from complexed enzymes by intramolecular electron transfer; consequently, all constructed CYTc-SC/ST-Enzyme complexes showed DET ability to the electrode, demonstrating the versatility of this method.
2023年02月, 研究論文（学術雑誌）, 共同, 24, 3, DOI(公開)(r-map) - Development of an Anti-Idiotype Aptamer-Based Electrochemical Sensor for a Humanized Therapeutic Antibody Monitoring
Nagata, Madoka; Lee, Jinhee; Saito, Taro; Ikebukuro, Kazunori; Sode, Koji
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
MDPI
Therapeutic monoclonal antibodies (mAbs) are currently the most effective medicines for a wide range of diseases. Therefore, it is expected that easy and rapid measurement of mAbs will be required to improve their efficacy. Here, we report an anti-idiotype aptamer-based electrochemical sensor for a humanized therapeutic antibody, bevacizumab, based on square wave voltammetry (SWV). With this measurement procedure, we were able to monitor the target mAb within 30 min by employing the anti-idiotype bivalent aptamer modified with a redox probe. A fabricated bevacizumab sensor achieved detection of bevacizumab from 1-100 nM while eliminating the need for free redox probes in the solution. The feasibility of monitoring biological samples was also demonstrated by detecting bevacizumab in the diluted artificial serum, and the fabricated sensor succeeded in detecting the target covering the physiologically relevant concentration range of bevacizumab. Our sensor contributes to ongoing efforts towards therapeutic mAbs monitoring by investigating their pharmacokinetics and improving their treatment efficacy.
2023年03月, 研究論文（学術雑誌）, 共同, 24, 6, DOI(公開)(r-map)
著書
- 和文論文 合計５報 和文総説 合計５編 和文著書 合計３編
2005年01月 - CHEMICAL, GAS, AND BIOSENSORS FOR INTERNET OF THINGS AND RELATED APPLICATIONS, Aptameric sensors utilizing its property as DNA
Kinuko Ueno, Kaori Tsukakoshi and Kazunori Ikebukuro
Elsevier
The biosensors using aptamer are reviewed and its cutting edge technology was explained.
2019年 - 「アプタマーの医療応用 」CSJカレントレビュー24 化学で医療・診断・創薬の革新を目指す
長谷川聖、池袋一典
医療･診断･創薬の化学 -医療分野に挑む革新的な化学技術-、日本化学会編、化学同人(2017)、p55-63.
2017年07月 - "History and Development of Nucleic Acid Aptamers" in Aptamers: Tools for Targeted Nanotherapy and Molecular ImagingApplications of Metaheuristics in Process Engineering,
Savory, N., Abe, K., Ikebukuro, K.
Pan Stanford publication
2015年11月 - In silico Maturation: Processing Sequences to Improve Biopolymer Functions Based on Genetic Algorithms, Applications of Metaheuristics in Process Engineering,
Savory, N., Abe, K., Yoshida, W., & Ikebukuro, K.
Springer
2014年02月 - 化学便覧応用化学編第7版 (2014/01)、8.5.4項「リボザイムとアプタマー」、
吉田亘、阿部公一、池袋一典
丸善出版株式会社
2014年01月 - “Electrochemical Biosensors Using Aptamers for Theranostics” in “Advances in Biochemical Engineering/Biotechnology”, 2013, Edited by Man Bock Gu,
Koichi Abe, Wataru Yoshida and Kazunori IKEBUKURO
Springer-Verlag
2014年01月 - 「バイオテクノロジー」、機械工学ハンドブック
池袋一典
朝倉書店
2011年07月 - “Aptamer sensors combined with enzymes for highly sensitive detection” in “Biosensors - Emerging Materials and Applications”
Koichi Abe and Kazunori IKEBUKURO
InTech
2011年07月 - Biosensors using Aptameric enzyme subunit; application of aptamers to allosteric control of enzymes” in “Analytial Applications of Aptamers”
Kazunori Ikebukuro, Wataru Yoshida, Koji Sode
Wiley
2009年01月 - バイオセンサ･ケミカルセンサ事典、「アプタマーセンサ」の項
軽部征夫監修
（株）テクノシステム
2007年07月 - 「アプタマーを分子認識素子とするバイオセンサの開発」 ユビキタスバイオセンシング
三林浩二監修
2006年01月 - 医学書院医学大辞典
共著者3400名超
医学書院
2003年03月 - “19. Cyanides” in “Handbook for Water Analysis”
共著者６０名
2000年10月 - Enzymes and Microbial Biosensors: Techniques and Protocols in “Methods in Biotechnology”, vol. 6
共著者４０名
1998年07月
研究発表、招待講演等
- “Computer aided evolution of the desired functions of peptide/DNA aptamers”
The third International Workshop on Symbiosis of Biology and Nanodevices
The computer aided evolution method for the function of aptamer is explained and some successful examples were shown.
2019年06月26日, 口頭発表（招待・特別） - Smart aptamer: the aptamer changing its structure and function depending on the environmental condition
Kazunori Ikebukuro
Smart aptamer sensing chemical environment and changing its structure and function was first reported and its application was shown.
2019年03月19日, 口頭発表（招待・特別） - 「グアニン四重鎖構造を形成するDNAのナノ構造変化を利用したセンシング」
日本学術振興会第174委員会第62回研究会
周囲の環境で構造と機能が変わるDNAアプタマーの紹介とこれを用いた応用について最先端の研究を紹介した。
2018年06月27日, 口頭発表（招待・特別） - BINDING PROPERTIES OF AMYLOID OLIGOMER-BINDING DNA APTAMERS AGAINST AMYLOID BETA OLIGOMERS
13th International Conference on Alzheimer’s and Parkinson’s Diseases (AD/PD 2017)
2017年03月29日, ポスター発表 - 標的DNA検出に向けたアルカリフォスファターゼ融合ジンクフィンガー蛋白質の開発
電気化学第84回大会
2017年03月25日, その他 - Development of the electrochemical detection system of thrombin activity
PepCon-2017
2017年03月22日, ポスター発表 - アミロイドオリゴマー結合アプタマーを用いた、核酸増幅に基づく水溶性アミロイドβオリゴマーの検出
日本化学会第97回春季年会
2017年03月16日, その他 - 非天然塩基の導入によるトロンビン結合アプタマーの結合能の改良
日本化学会第97回春季年会
2017年03月16日, その他 - シトシンメチル化がG-quadruplexとタンパク質の結合に及ぼす影響の評価
第39回 日本分子生物学会年会
2016年12月01日, ポスター発表 - 抗体医薬品を特異的に認識する抗イディオタイプアプタマーの探索とその特性評価
第39回 日本分子生物学会年会
2016年12月01日, ポスター発表 - Development of the Electrochemical Detection System Using the Combination of Aptamer and Enzyme
PRiME 2016
2016年10月02日, 口頭発表（一般） - Development of a detection system for epigenetic modifications using enzyme fused zinc finger protein
PRiME 2016
2016年10月02日, 口頭発表（一般） - Construction of photo regulation system of protein expression in Synechocystis sp. PCC 6803
The 43rd International Symposium on Nucleic Acids Chemistry.
2016年09月28日, ポスター発表 - Effect of G-quadruplex ligand on the topology of G-quadruplex forming aptamer and its affinity to the target molecules
The 43rd International Symposium on Nucleic Acids Chemistry.
2016年09月28日, ポスター発表 - Screening and characterization of aptamer for myoglobin
The 43rd International Symposium on Nucleic Acids Chemistry.
2016年09月28日, ポスター発表 - Development of DNA aptamers against FokI nuclease domain for genome editing
The 43rd International Symposium on Nucleic Acids Chemistry.
2016年09月28日, ポスター発表 - グアニン四重鎖特異的リガンドを用いたDNAアプタマーの構造制御
第10回バイオ関連化学シンポジウム
2016年09月08日, 口頭発表（一般） - 高感度検出への応用を目指したアルカリホスファターゼ阻害アプタマーの探索
第10回バイオ関連化学シンポジウム
2016年09月08日, ポスター発表 - リボレギュレーターを用いた内在性遺伝子cyabrB2 の転写および発現制御
第10回バイオ関連化学シンポジウム
2016年09月08日, ポスター発表 - タンパク質とG-quadruplexとの結合にシトシンのメチル化が与える影響の解析
第10回バイオ関連化学シンポジウム
2016年09月08日, ポスター発表 - Improvement of binding affinity of G-quadruplex forming aptamers
Biosensor2016
2016年05月26日, ポスター発表 - Interaction between G-quadruplex (G4)-forming aptamer and heme protein
Biosensor2016
2016年05月26日, ポスター発表 - Development of a detection system for epigenetic modifications using enzyme fused zinc finger protein
Biosensor2016
2016年05月26日, 口頭発表（一般） - Selection and characterization of DNA aptamers against FokI nuclease domain for genome editing
Biosensor2016
2016年05月26日, 口頭発表（一般） - Development of multivalent aptamers for high-sensitive detection of target proteins
Biosensor2016
2016年05月26日, 口頭発表（一般） - シトシンのメチル化がG-quadruplex構造とタンパク質との結合に及ぼす影響の解析
日本化学会第96回春季年会
2016年03月26日, その他 - Synechocystis sp. PCC 6803におけるリボレギュレーターを用いた転写因子cyAbrB2の発現制御
日本化学会第96回春季年会
2016年03月26日, その他 - Development of Light inducible riboregulator for Escherichia coli
2nd International Workshop on Cyanofactory
2016年03月05日, ポスター発表 - Screening method to obtain aptamers binding different sites of the target molecule
Pacifichem 2015
2015年12月17日, ポスター発表 - Characterization of aptamer forming G-quadruplex structure and its binding to target molecules in hydrated ionic liquid
Pacifichem 2015
2015年12月17日, ポスター発表 - Investigation of the effects of G-quadruplex ligand (7OTD) on the expression of cell transcriptome
Pacifichem 2015
2015年12月16日, ポスター発表 - “RNA-based gene regulators for controlling gene expression in cyanobacteria”
Pacifichem 2015
2015年12月16日, その他 - Engineering of a wide dynamic range green-light regulation system for gene expression in Synechocystis sp. PCC 6803
Pacifichem 2015
2015年12月16日, ポスター発表 - G-quadruplex (G4) 形成アプタマーの構造制御及び結合能向上に向けたG4 リガンドの応用
第38回 日本分子生物学会年会・第88回日本生化学会大会 合同大会
2015年12月03日, ポスター発表 - ゲノム編集技術への応用を目指したFokIヌクレアーゼドメインに結合するDNAアプタマーの探索及び特性評価
第38回 日本分子生物学会年会・第88回日本生化学会大会 合同大会
2015年12月03日, ポスター発表 - Identification of DNA aptamers from G-quadruplex sequence in human genome”
1st International Symposium on Functional Nucleic Acids: From Laboratory to Targeted Molecular Therapy”
2015年11月27日, 口頭発表（一般） - 「シアノバクテリアで機能するリボレギュレーターのエンジニアリング」
藍藻の分子生物学2015
2015年10月27日, 口頭発表（招待・特別） - Synechocystis sp. PCC 6803由来RpaBの発現を抑制する人工small RNAの開発」
藍藻の分子生物学2015
2015年10月27日, ポスター発表 - 二次構造予測に基づくsmall RNAの機能改良法の開発」
第67回日本生物工学会大会
2015年10月27日, ポスター発表 - FokIヌクレアーゼドメインに対するDNAアプタマーの構造及び機能解析
第67回日本生物工学会
2015年10月27日, ポスター発表 - G-quadruplex (G4) 形成アプタマーの構造及び結合能に対するG4リガンドの影響の解析
第67回日本生物工学会
2015年10月27日, ポスター発表 - Analysis of interaction between aptamers and target molecules in hydrated ionic liquid
The 42nd International Symposium on Nucleic Acids Chemistry.
2015年09月24日, ポスター発表 - Detection of anti-VEGF antibody bevacizumab using DNA aptamer
The 42nd International Symposium on Nucleic Acids Chemistry.
2015年09月24日, ポスター発表 - 熱応答性磁性ナノ粒子と DNA アプタマーを用いた 標的タンパク質検出システムの開発
pre-mRNA中に存在するG-quadruplex構造形成配列に着目したアプタマー探索
2015年09月11日, 口頭発表（一般） - 熱応答性磁性ナノ粒子と DNA アプタマーを用いた 標的タンパク質検出システムの開発
第9回バイオ関連化学シンポジウム
2015年09月11日, ポスター発表 - FokIヌクレアーゼドメインに対するDNAアプタマーの探索
第9回バイオ関連化学シンポジウム
2015年09月11日, ポスター発表 - G4リガンドがVEGF結合アプタマーの構造及び結合能に与える影響の解析
第9回バイオ関連化学シンポジウム
2015年09月11日, ポスター発表 - 水和イオン液体中でのアプタマーと標的分子の相互作用解析
2015年電気化学秋季大会
2015年09月11日, その他 - Investigation of the effect of G-quadruplex ligand on the expression and splicing of VEGF mRNA.
5th International Meeting on Quadruplex Nucleic Acids: G4thering in Bordeaux
2015年05月27日, ポスター発表 - 大腸菌で機能する光誘導型リボレギュレーターシステムの開発
日本化学会第95回春季年会
2015年03月26日, その他 - 水和イオン液体中でのトロンビン-トロンビン結合アプタマーの結合
日本化学会第95回春季年会
2015年03月26日, その他 - 抗VEGF抗体bevacizumab(Avastin) のanti-idiotype aptamerの探索
日本化学会第95回春季年会
2015年03月26日, その他 - Synechocystis sp. PCC6803内における緑色光誘導型リボレギュレーターの特性評価
日本化学会第95回春季年会
2015年03月26日, その他 - DNA 四重鎖構造の安定性に着目したメチル化 DNA 簡易検出法の開発
日本化学会第95回春季年会
2015年03月26日, その他 - Biosensing based on the change of nano-structure of aptamer by Kazunori Ikebukuro
Sweden-Japan seminar on Nanomaterials and Nanotechnology
2015年03月11日, 口頭発表（招待・特別） - in silico maturation 法を用いたアプタマーの機能改良
第 10 回理研「バイオものづくり」シンポジウム
2015年03月06日, 口頭発表（招待・特別） - Engineering artificIn silico maturation: improvement of aptamer function based on genetic algorithms employing in vitro functional screening process
The 41th International Symposium on Nucleic Acids Chemistry
2014年11月05日, 口頭発表（一般） - Engineering artificial small RNAs to control post-transcriptional gene expression in Synechocystis sp. PCC6803
The 41th International Symposium on Nucleic Acids Chemistry
2014年11月05日, ポスター発表 - グルコース脱水素酵素融合ジンクフィンガー蛋白質の電気化学検出への応用
2014年度電気化学会秋季大会
2014年09月27日, その他 - RNA結合タンパク質Hfqを利用したSynechocystis sp. PCC 6803内で機能するリボレギュレーターの遺伝子発現制御能の向上
第8回バイオ関連化学シンポジウム
2014年09月11日, 口頭発表（一般） - Engineering the scaffold region of natural and artificial small RNAs to enhance their gene regulation abilities PCC6803
RNA2014
2014年06月05日, ポスター発表 - DNA aptamers against the Cry j 2 allergen of Japanese cedar pollen
Biosensor2014
2014年05月28日, ポスター発表 - In silico maturation: Advanced aptamer development for biosensing applications
Biosensor2014
2014年05月28日, ポスター発表 - RNA aptamer screening focusing on G-quadruplex forming region of pre-mRNA
Biosensor2014
2014年05月28日, ポスター発表 - Detection of epigenetic modifications using enzyme fused zinc finger protein
Biosensor2014
2014年05月28日, ポスター発表 - Screening and improvement of DNA aptamers by in silico approachesspecificity
Biosensor2014
2014年05月28日, ポスター発表 - A novel RNA based genetic control system for metabolic gene regulation PCC6803
4th International Conference on Algal Biomass, Biofuels & Bioproducts
2014年05月15日, その他 - Loop-engineering of riboregulator to improve its gene regulation in Synechocystis sp. PCC 6803
The 10th Asia-Pacific Marine Biotechnology Conference
2014年05月05日, その他 - 櫛型電極を用いた電気化学的なトロンビン活性の検出法の開発
電気化学会第82回大会
2014年03月16日, その他 - 櫛型電極を用いた電気化学的なトロンビン活性の検出法の開発
電気化学会第82回大会
2014年03月15日, その他 - スギ花粉アレルゲンCry j 2に結合するDNAアプタマーの開発
第36回日本分子生物学会年会
2013年12月03日, ポスター発表 - Aptamer selection based on putative G-quadruplexsequence in genome
The 40th International Symposium on Nucleic Acids Chemistry
2013年11月14日, 口頭発表（一般） - Screening and Improvement of DNA Aptamers against Hepatocyte Growth Factor by in silico Approaches
The 40th International Symposium on Nucleic Acids Chemistry
2013年11月13日, ポスター発表 - Detection of DNA methylation and histone modification by enzyme fused zinc finger protein
The 40th International Symposium on Nucleic Acids Chemistry
2013年11月13日, ポスター発表 - Improving gene regulation ability of bacterial small RNAs by scaffold engineering
The 40th International Symposium on Nucleic Acids Chemistry
2013年11月13日, ポスター発表 - 酵素融合ジンクフィンガー蛋白質を用いた遺伝子検出システムの開発
電気化学会秋季大会
2013年09月27日, その他 - Zinc-finger nucleaseを細胞内で特異的に放出する表面修飾法の検討
第7回バイオ関連化学シンポジウム
2013年09月26日, ポスター発表 - Hfq結合配列の改良によるsmallRNAの遺伝子発現制御能の向上
第7回バイオ関連化学シンポジウム
2013年09月26日, その他 - In silico 配列解析及び操作によるアプタマーの探索と改良
第7回バイオ関連化学シンポジウム
2013年09月26日, ポスター発表 - ループ領域改変によるリボレギュレーターの遺伝子発現能の向上
第7回バイオ関連化学シンポジウム
2013年09月26日, ポスター発表 - リボレギュレーターの光制御
第7回バイオ関連化学シンポジウム
2013年09月26日, ポスター発表 - 藍藻で機能するリボレギュレータの改変
第65回日本生物工学会大会
2013年09月18日, 口頭発表（一般） - ゲノム中のG-quadruplex形成配列に着目したアプタマー探索法
第65回日本生物工学会大会
2013年09月18日, ポスター発表 - Identification of HGF and HBEGF aptamers by “G4 Promoter-derived Aptamer Selection (G4PAS)”
The Sixth International Meeting on Synethetic Biology
2013年07月10日, ポスター発表 - Engineering Hfq binding sequence to control gene regulation of bacterial small RNA
The Sixth International Meeting on Synethetic Biology
2013年07月10日, ポスター発表 - Selection of DNA aptamers targeted to Proteus mirabilis membrane-associated proteins.
SfAM Summer Conference 2013
2013年07月04日, ポスター発表 - Aptamer selection based on genomic information; focusing on G-quadruplex forming sequence
4th International Meeting on Quadruplex Nucleic Acids
2013年07月04日, ポスター発表 - 微生物ゲノムDNAの電気化学的検出法に向けたGDH融合ジンクフィンガー蛋白質の開発
電気化学会創立第80周年記念大会
2013年03月30日, その他 - アプタマーとペプチド核酸を用いた血管内皮細胞増殖因子の検出システムの開発
電気化学会創立第80周年記念大会
2013年03月30日, その他 - ノロウイルスの外殻タンパク質VP1に結合するDNAアプタマーの探索
日本化学会第94回春季年会
2013年03月27日, その他 - Synechocystis sp. PCC6803において高い遺伝子発現制御能を持つリボレギュレータへの改良
日本化学会第94回春季年会
2013年03月27日, その他 - GDH 融 合 ジンクフィンガー蛋白質を用いた微生物ゲノムの電気化学的検出法の開発
日本化学会第94回春季年会
2013年03月25日, その他 - ループ構造改変によるリボレギュレータの改良
日本化学会第94回春季年会
2013年03月25日, その他 - RNA結合蛋白質を利用したリボレギュレータの改良
日本化学会第94回春季年会
2013年03月25日, 口頭発表（一般） - αシヌクレインの凝集線維化におけるランタニド系金属の影響
日本化学会第94回春季年会
2013年03月24日, その他 - CpGアイランドから見出されたグアニン四重鎖形成配列と低分子化合物の相互作用解析
日本化学会第94回春季年会
2013年03月24日, その他 - 特 異 的 蛍 光 リガンドと DNA マイクロアレイを基盤とした新規グアニン四重鎖形成配列の網羅的検出法の開発
日本化学会第94回春季年会
2013年03月24日, 口頭発表（一般） - ゲノム中のG-quadruplex形成配列情報に基づいたバイオマーカー結合アプタマーの探索
日本化学会第94回春季年会
2013年03月24日, その他 - ゲノム中のG-quadruplex構造形成配列に着目したアプタマー探索法の開発
日本化学会第94回春季年会
2013年03月24日, その他 - 二次元電気泳動装置システムを利用した、アプタマーのスクリーニング方法の開発
第35回日本分子生物学会年会
2012年12月13日, ポスター発表 - VEGF detection using aptamer and PNA-based Bound/Free separation system
The 39th International Symposium on Nucleic Acids Chemistry
2012年11月16日, ポスター発表 - Post-transcriptional gene regulation by artificial riboregulator in cyanobacteria
The 39th International Symposium on Nucleic Acids Chemistry
2012年11月16日, ポスター発表 - In silico selection of aptamers based on genomic information
The 39th International Symposium on Nucleic Acids Chemistry
2012年11月16日, その他 - アミロイドオリゴマー結合アプタマーの Aβオリゴマーに対する認識特異性
認知症学会
2012年10月26日, ポスター発表 - リボレギュレーターによるシアノバクテリアの遺伝子発現制御
第64回日本生物工学会大会
2012年10月24日, その他 - 遺伝子的アルゴリズムを適用したompCプロモーターの改良
第64回日本生物工学会大会
2012年10月24日, その他 - ルシフェラーゼ融合ジンクフィンガータンパク質を用いたヒストン修飾解析法の開発
第64回日本生物工学会大会
2012年10月24日, その他 - Cell-SELEXによる尿路感染症起因菌に結合するDNAアプタマーの探索
第64回日本生物工学会大会
2012年10月24日, その他 - 藍藻で機能するリボレギュレータのデザイン
第64回日本生物工学会大会
2012年10月24日, その他 - Aptameric Sensor for Detection of VEGF Based on Labeling Technique Using GDH Fused Zinc Finger Protein
電気化学会秋季大会（Prime2012)
2012年10月11日, その他 - VEGF結合アプタマーのin silico maturation
第6回バイオ関連化学シンポジウム2012
2012年09月26日, ポスター発表 - アミロイドオリゴマー結合DNAアプタマーのオリゴマー結合特異性の解析
第6回バイオ関連化学シンポジウム2012
2012年09月26日, ポスター発表 - In silico アプタマー探索法の開発
第6回バイオ関連化学シンポジウム2012
2012年09月26日, その他 - 二次元電気泳動装置「Auto2D」を利用した、DNAアプタマーのスクリーニング方法の開発
日本プロテオーム学会2012年大会
2012年07月26日, ポスター発表 - Construction of the Autoaggregation and Autolysis System Controlled by Riboregulator
The 9th Asia-Pacific Marine Biotechnology Conference
2012年07月13日, その他 - Directed evolution of ompC promoter by applying genetic algorithm
The 9th Asia-Pacific Marine Biotechnology Conference
2012年07月13日, その他 - Design of Riboregulators that Function in Cyanobacteria
The 9th Asia-Pacific Marine Biotechnology Conference
2012年07月13日, その他 - Construction of novel RNA tools for controlling synthetic cyanobacterial bioprocess
The 9th Asia-Pacific Marine Biotechnology Conference
2012年07月13日, その他 - Construction of the autoaggregation and autolysis system controlled by riboregulator
The 2nd International Conference on Algal Biomass, Biofuels and Bioproducts
2012年06月10日, その他 - Design of riboregulators that function in cyanobacteria
The 2nd International Conference on Algal Biomass, Biofuels and Bioproducts
2012年06月10日, その他 - Cell-SELEX for selection of anti-Proteus mirabilis DNA aptamers and in silico maturation to improve binding-specificity
Biosensor2012
2012年05月15日, その他 - Improvement of Affinities of VEGF-binding Aptamers for Biosensor by in silico maturation
Biosensor2012
2012年05月15日, その他 - Development of DNA aptamers that specifically bind to amyloid protein oligomer and flow cytometric detection of amyloid beta oligomer
Biosensor2012
2012年05月15日, その他 - Quantitaive DNA methylation level measurement using MBD and Zinc finger fused luciferase
Biosensor2012
2012年05月15日, その他 - Riboregulator for controlling synthetic cyanobacterial bioprocess
Biosensor2012
2012年05月15日, その他 - セラノスティックスとバイオセンサー」
特別企画「生物電気化学・バイオエンジニアリング領域の新展開」、電気化学会第79回大会
2012年03月31日, 口頭発表（招待・特別） - 微生物ゲノムの電気化学的検出法に向けたGDH融合ジンクフィンガー蛋白質の開発
電気化学会創立第80周年記念大会
2012年03月29日, その他 - アプタマーとペプチド核酸を用いた血管内皮細胞増殖因子の検出システムの開発
電気化学会創立第80周年記念大会
2012年03月29日, その他 - GDH 融合ジンクフィンガー蛋白質を用いた微生物ゲノムの電気化学的検出法の開発
日本化学会第93回春季年会
2012年03月23日, その他 - ループ構造改変によるリボレギュレータの改良
日本化学会第93回春季年会
2012年03月23日, その他 - ゲノム中の G-quadruplex 形成配列情報に基づいたバイオマーカー結合アプタマーの探索
日本化学会第93回春季年会
2012年03月23日, その他 - ゲノム中の G-quadruplex 構造形成配列に着目したアプタマー探索法の開発
日本化学会第93回春季年会
2012年03月23日, その他 - RNA 結合蛋白質を利用したリボレギュレータの改良
日本化学会第93回春季年会
2012年03月23日, その他 - 「アミロイドオリゴマーを認識する核酸リガンドDNAアプタマーの開発」
トピックス討論「認知症基礎研究のホットトピックス徹底討論」、第30回日本認知症学会学術集会
2011年11月12日, 口頭発表（招待・特別） - Alpha fetoproteinに結合するDNAアプタマーの探索とその改良
第63回日本生物工学会大会
2011年09月26日, その他 - ジンクフィンガー蛋白質融合ルシフェラーゼを用いた複数病原性微生物の検出システムの開発
第63回日本生物工学会大会
2011年09月26日, その他 - 磁性ビーズを用いたDNAメチル化レベル評価法の開発
第63回日本生物工学会大会
2011年09月26日, その他 - In silico maturation法を用いたVEGF結合アプタマーの改良
第63回日本生物工学会大会
2011年09月26日, その他 - EcoTanker - 光アクチュエーターで操縦する大腸菌・マイクロ・タンカー
第63回日本生物工学会大会
2011年09月26日, その他 - Proteus mirabilisに結合するDNAアプタマーの探索及びin silico maturation法による特異性の改良
第63回日本生物工学会大会
2011年09月26日, その他 - Detection of VEGF using aptamer labeled with FADGDH fused zinc finger protein
62nd Annual Meeting of the International Society of Electrochemistry
2011年09月14日, その他 - IN SILICO MATURATION OF VEGF-BINDING APTAMER FOR ELECTROCHEMICAL VEGF-DETECTION SYSTEM WITH APTAMER SANDWICH
62nd Annual Meeting of the International Society of Electrochemistry
2011年09月12日, ポスター発表 - VEGF検出用Capturable aptamerのin silico maturationによる改良
2011年 電気化学秋季大会
2011年09月11日, その他 - GDH標識アプタマーを用いたVEGFの電気化学的検出
2011年 電気化学秋季大会
2011年09月11日, その他 - Development of DNA aptamers against Proteus mirabilis by whole-cell SELEX and in silico maturation
International Union of Microbiological Societies 2011 Congress
2011年09月07日, ポスター発表 - Selection of DNA aptamers against Proteus mirabilis using whole-cell SELEX
Society for General Microbiology, Spring Conference 2011
2011年04月14日, ポスター発表 - In silico maturationによるVEGF結合DNAアプタマーの改良
電気化学会第78回大会
2011年03月29日, その他 - GDH標識アプタマーの構造変化を利用したVEGFの電気化学的測定
電気化学会第78回大会
2011年03月29日, その他 - アミロイド蛋白質オリゴマーに結合するDNA アプタマーの開発と応用
日本化学会第91回春季年会
2011年03月29日, その他 - キメラEnvZ/OmpR系Two-somponent systemを用いた転写制御系
日本化学会第91回春季年会
2011年03月29日, その他 - GDH標識DNAアプタマーを用いたVEGF検出系の開発
日本化学会第91回春季年会
2011年03月29日, その他 - Alpha-fetoproteinに結合するDNAアプタマーの探索
日本化学会第91回春季年会
2011年03月29日, その他 - ジンクフィンガー融合ルシフェラーゼを用いた病原性微生物の自動検出法の開発
日本化学会第91回春季年会
2011年03月29日, その他 - 磁性ビーズを用いたDNAメチル化レベル評価法の開発
日本化学会第91回春季年会
2011年03月29日, その他 - VEGFに結合するDNAアプタマーの探索と疾病診断用センサー素子としての改良
日本化学会第91回春季年会
2011年03月29日, その他 - 進化分子工学を利用した新規分子認識素子の開発
東京バイオマーカー・イノベーション技術研究組合、第1回研究交流フォーラム
2011年03月08日, 口頭発表（招待・特別） - メチル化CpG 結合タンパク質を用いた新規メチル化頻度評価法の開発3
第62回日本生物工学会大会
2010年10月29日, ポスター発表 - アプタマー酵素サブユニット(AES)を用いた高感度疾患マーカーの検出」
シンポジウムⅠ ナノテクノロジーが拓く未来の臨床検査、日本臨床検査自動化学会第42回大会
2010年10月09日, 口頭発表（招待・特別） - アプタマーの進化法：in silico maturation
イノベーション･ジャパン2010 新技術説明会
2010年10月01日, 口頭発表（一般） - Aptameric sensors based on structural change for diagnosis
Faraday discussion 149:Analysis for Healthcare Diagnostics and Theranostics
2010年09月07日, 口頭発表（一般） - アプタマーを用いた電気化学的VEGF測定法の開発
2010年電気化学秋季大会
2010年09月03日, その他 - C反応性蛋白質に結合するアプタマーの改良
2010年電気化学秋季大会
2010年09月03日, その他 - 前立腺特異抗原(PSA)に結合するDNAアプタマーの探索及び改良
2010年電気化学秋季大会
2010年09月03日, その他 - 細胞障害性オリゴマー検出に向けたαシヌクレイン結合DNAアプタマーの探索
2010年電気化学秋季大会
2010年09月02日, その他 - 疾患マーカーを特異的に認識するアプタマーの開発とセンシングへの応用
新技術説明会
2010年06月22日, 口頭発表（一般） - Structural control of aptamer inserted polymerized G-quadruplex DNA nanostructure (Gq-switch) by small molecule and its application to biosensing
Biosensors 2010
2010年05月28日, 口頭発表（一般） - The development of an autonomous self-powered bio-sensing actuator
Biosensors 2010
2010年05月27日, 口頭発表（基調） - The development of a novel theranostic platform for cytotoxicity evaluation of amyloid-forming neurodegenerative disease causative proteins
Biosensors 2010
2010年05月26日, 口頭発表（一般） - AptaDevelopment of aptameric sensor using conformational change of glucosedehydrogenase binding aptamer
Biosensors 2010
2010年05月26日, ポスター発表 - 酵素融合ジンクフィンガー蛋白質を用いた標的分子検出法の開発
日本化学会第90春季年会
2010年03月29日, その他 - 染色体DNAの操作を目的としたテロメスタチン誘導体とヒトテロメア配列の力学的相互作用解析
日本化学会第90春季年会
2010年03月29日, その他 - メチル化DNA結合タンパク質MBD1を用いた新規メチル化頻度測定系の開発
日本化学会第90春季年会
2010年03月29日, その他 - アミロイド蛋白質細胞毒性バイオセンシングのセラノスティスクへの応用
日本化学会第90春季年会
2010年03月28日, その他 - 腫瘍マーカー検出用DNAアプタマーの探索
日本化学会第90春季年会
2010年03月28日, その他 - 前立腺特異抗原（PSA）に結合するDNAアプタマーの探索
日本化学会第90春季年会
2010年03月28日, その他 - センサー素子にアプタマーを用いた高感度測定系の構築を目的としたVEGFアプタマーの改良
2009年電気化学会秋季大会
2009年09月11日, その他 - ホタル由来ルシフェラーゼに結合するDNAプタマーの探索
2009年電気化学会秋季大会
2009年09月11日, その他 - ルシフェラーゼ融合ジンクフィンガーを用いたサルモネラ属の検出
2009年電気化学会秋季大会
2009年09月11日, その他 - C-reactive protein及びthyroglobulinに結合するDNAアプタマーの探索と改良
2009年電気化学会秋季大会
2009年09月11日, その他 - PQQGDHアプタマーを用いたaptamer sensorの開発
2009年電気化学会秋季大会
2009年09月11日, その他 - ジンクフィンガー蛋白質を用いた機能性蛋白質と機能性DNA分子の融合
2009年電気化学会秋季大会
2009年09月10日, その他 - α-シヌクレインに結合するDNAアプタマーの探索と測定への応用
2009年電気化学会秋季大会
2009年09月10日, その他 - Biosensors Based on Aptamer's Structural Change
AsiaSense2009, July 31st, 2009,
2009年07月31日, 口頭発表（基調） - Selection of DNA aptamers agaianst firefly luciferase
First World Congress of International Academy of Nanomedicine(IANM), June 12, 2009, Hinan, China
2009年06月12日, ポスター発表 - Improvement of DNA aptamers against C-reactive protein and thyroglobulin
First World Congress of International Academy of Nanomedicine(IANM), June 12, 2009, Hinan, China
2009年06月12日, ポスター発表 - Detection method of PCR products from Salmonella using fusion protein of zinc finger protein & firefly luciferase
First World Congress of International Academy of Nanomedicine(IANM), June 12, 2009, Hinan, China
2009年06月12日, ポスター発表 - Aptamers and zinc finger proteins as nanotools for diagnosis
First World Congress of International Academy of Nanomedicine(IANM), June 12, 2009, Hinan, China
2009年06月12日, 口頭発表（招待・特別） - 蛍光偏向解消法を用いたαシヌクレインオリゴマーの検出
蛋白質科学会
2009年05月22日, ポスター発表 - ルシフェラーゼ融合ジンクフィンガー蛋白質を用いたアプタマー標識法の開発
日本化学会第89春季年会
2009年03月28日, 口頭発表（一般） - 血管内皮細胞増殖因子(VEGF165)を認識するDNAアプタマーの改良
日本化学会第89春季年会
2009年03月28日, 口頭発表（一般） - アミロイド形成タンパク質の新規細胞毒性バイオセンシング系の開発
日本化学会第89春季年会
2009年03月28日, 口頭発表（一般） - αシヌクレインに結合するDNAアプタマーの探索
日本化学会第89春季年会
2009年03月28日, 口頭発表（一般） - アミロイド形成蛋白質αシヌクレイン中に見出される繰り返し配列の構造形成における役割
日本化学会第89春季年会
2009年03月28日, 口頭発表（一般） - Thyroglobulin に結合するDNA アプタマーの改良の探索
日本化学会第89春季年会
2009年03月28日, 口頭発表（一般） - C-reactive proteinに結合するDNAアプタマーの探索
日本化学会第89春季年会
2009年03月28日, 口頭発表（一般） - Thyroglobulinに結合するDNAアプタマーの探索および改良
日本分子生物学会
2008年12月09日, ポスター発表 - 酵素センサーへの応用を目指したFADGDHに結合するDNAアプタマーの探索
日本分子生物学会
2008年12月09日, ポスター発表 - C-reactive proteinに結合するDNAアプタマーの探索および改良
日本分子生物学会
2008年12月09日, ポスター発表 - BioCapacitor ~ A stand alone, self-powered wireless glucose sensing system~ and Firefly Luciferase fusion protein
PACIFIC RIM MEETING 2008 on electrochemical and solid-state science
2008年10月16日, その他 - FADグルコース脱水素酵素の遺伝的アルゴリズムによる基質特異性の改良
日本生物工学会
2008年08月28日, ポスター発表 - Detection of PCR Products from Legionella pneumophila using Zinc Finger Protein and Firefly Luciferase fusion protein
Biosensors 2008, THe 10th World congress on Biosensors
2008年05月14日, その他 - BioCapacitor ~ A Novel Category Of Biosensor ~
Biosensors 2008, THe 10th World congress on Biosensors
2008年05月14日, その他 - Bound/Free separation based on structural change of aptamers and its application to biosensing
Biosensors 2008, THe 10th World congress on Biosensors
2008年05月14日, その他 - The Novel Detection System of PCR Products from Bacterial Genome using Zn Finger Proteins
Biosensors 2008, THe 10th World congress on Biosensors
2008年05月14日, その他 - Aptameric enzyme subunit;novel recognition biosensing element for allosteric enzymecontrol
Biosensors 2008, THe 10th World congress on Biosensors
2008年05月14日, 口頭発表（基調） - ルシフェラーゼに結合するDNA アプタマーの探索
日本化学会第88春季年会
2008年03月27日, その他 - Thyroglobulinに結合するDNAアプタマーの探索Aptameric Enzyme Subunit への応用を目指したFADGDH 活性に影響を及ぼすDNA アプタマーの探索
日本化学会第88春季年会
2008年03月27日, その他 - Thyroglobulinに結合するDNAアプTau タンパク質に結合するDNA アプタマーのスクリーニングタマーの探索
日本化学会第88春季年会
2008年03月27日, その他 - 標的分子認識に伴い構造変化するアプタマーのデザインとそのB/F分離への応用ルシフェラーゼに結合するDNA アプタマーの探索
日本化学会第88春季年会
2008年03月27日, その他 - 標的分子認識に伴い構造変化するアプタマーのデザインとそのB/F分離への応用
日本化学会第88春季年会
2008年03月27日, その他 - C-reactive proteinに結合するDNAアプタマーの探索
日本化学会第88春季年会
2008年03月27日, その他 - Thyroglobulinに結合するDNAアプタマーの探索
日本化学会第88春季年会
2008年03月27日, その他 - APTAMERIC ENZYME SUBUNIT (AES) BASED BIOSENSING
THIRD INTERNATIONAL WORKSHOP ON "BIOSENSORS FOR FOOD SAFETY AND ENVIRONMENTAL MONITORING"
2007年10月07日, 口頭発表（一般） - Homogeoeous protein detection using aptamer based on laser interferometric photo-thermal displacement measurement
10th International Conference BIODETECTION TECHNOLOGIES 2007
2007年10月05日, ポスター発表 - Detection of interaction between aptamer and protein based on laser interferometric photo-thermal displacement measurement
3rd Annual Meeting of the Oligonucleotide Therapeutics Society
2007年10月05日, ポスター発表 - Development of simple and rapid detection method of pathogenic bacteria using Zn finger protein
10th International Conference BIODETECTION TECHNOLOGIES 2007
2007年07月14日, ポスター発表 - アプタマーを用いたバイオセンシング法の開発
第5回 細胞工学技術研究会
2007年07月09日, 口頭発表（招待・特別） - Aptameric biosensors
Japan- American Frontier of Science symposium
2006年12月08日, 口頭発表（一般） - Screening of peptide ligands by in silico panning
Japan-China-Korea Joint Symposium on Enzyme Engineering
2006年10月30日, 口頭発表（一般） - Decreasing the fibrillation and cytotoxicity of alpha-synuclein, a causative factor of Parkinson's disease
Japan-China-Korea Joint Symposium on Enzyme Engineering
2006年10月30日, 口頭発表（一般） - Aptameric enzyme subunit； novel analytical nanotool
2nd ICBN int'l conference of Bio-nanointerface
2006年10月10日, ポスター発表 - 電磁場による金ナノ粒子修飾トロンビンアプタマーの機能制御
第９回日本化学会バイオテクノロジー部会シンポジウム
2006年09月28日, ポスター発表 - マウスプリオンアプタマーの探索とそのセンシングへの応用
第９回日本化学会バイオテクノロジー部会シンポジウム
2006年09月28日, 口頭発表（一般） - DNA固定化担体によりB/F分離を行うタンパク質検出用アプタマーセンサーの開発
第９回日本化学会バイオテクノロジー部会シンポジウム
2006年09月28日, 口頭発表（一般） - Znフィンガータンパク質を用いた病原性微生物の特異的検出法の開発
第58回日本生物工学会大会
2006年09月11日, その他 - ピロロキノリンキノンによるヒト由来alpha-シヌクレインの凝集・線維化への影響
第58回日本生物工学会大会
2006年09月11日, その他 - Evolution of thrombin-inhibiting DNA aptamers based on inhibitory activity using a genetic algorithm
XVIIinternational roundtable on Nucleosides, Nucleotides and Nucleic acids
2006年09月05日, ポスター発表 - Aptameric enzyme subunit as novel molecular element for biosensing
XVIIinternational roundtable on Nucleosides, Nucleotides and Nucleic acids
2006年09月05日, ポスター発表 - Aptameric biosensors; novel sensing systems based on aptamers’ properties
3rd International Symposium on Innovative BioPhysio Sensor Technology, July 7, 2006, Jeju
2006年07月07日, 口頭発表（招待・特別） - Aptameric enzyme subunit: Novel biosensing element based on enzyme-inhibiting aptamer
Biosensor 2006
2006年05月09日, ポスター発表 - Development of aptamer based protein detection system using DNA immobilized support for bound/free separation
Biosensor 2006
2006年05月09日, 口頭発表（一般） - 国際学会口頭発表２５件以上
2006年04月, その他 - 国内学会口頭発表 ２５件以上
2006年04月, その他 - DNA固定化担体を用いた電気化学的タンパク質検出システムの開発
電気化学会第73回大会
2006年04月02日, 口頭発表（一般） - 金属ナノ粒子修飾トロンビン阻害DNAアプタマーの作製と特性検討
電気化学会第73回大会
2006年04月02日, 口頭発表（一般） - 電位を印加することによるa-シヌクレインの凝集の加速および凝集体の評価
電気化学会第73回大会
2006年04月02日, 口頭発表（一般） - ジンクフィンガー蛋白質Sp2を用いたLegionella pneumophila strain philadelphia 1の迅速・簡便な検出法の開発
日本化学会第86春期年会
2006年03月27日, 口頭発表（一般） - 変異αシヌクレインの細胞毒性評価
日本化学会第86春期年会
2006年03月27日, 口頭発表（一般） - 複数標的蛋白質に対するDNAアプタマーの同時探索法の開発
日本化学会第86春期年会
2006年03月27日, 口頭発表（一般） - Aptamer blottingによる細胞中の蛋白質に対するDNAアプタマーの探索（２）
日本化学会第86春期年会
2006年03月27日, 口頭発表（一般） - Aptamer blottingによる細胞中の蛋白質に対するDNAアプタマーの探索（１）
日本化学会第86春期年会
2006年03月27日, 口頭発表（一般） - Aptameric enzyme subunit； novel analytical nanotool
NSF/MEXT U.S. Young scientists symposium on bionanotechnology
2005年12月09日, 口頭発表（招待・特別） - Aptameric enzyme subunit as a novel nano sensing probe
NSF/MEXT U.S. Young scientists symposium on bionanotechnology
2005年03月15日, 口頭発表（招待・特別） - Technology transfer from the University to the industry
6th Japan-EU biotechnology policy working group meeting (Brussels, Belgium)
2001年02月12日, 口頭発表（招待・特別） - 迅速測定用DNAセンサーの開発
計測連合シンポジウム 先端計測2000
2000年05月18日, 口頭発表（招待・特別） - Biosensors for environmental monitoring
An approarch to environmental sensing, Biomolecular mechanism & design workshop '99 (Tsukuba, Japan)
1999年06月, 口頭発表（招待・特別） - Recent achievements in biosensors development in Japan
New trends in biosensor development, NATO advanced research workshop (Kiev, Ukraine)
1998年07月, 口頭発表（招待・特別） - Molecular imprinting technique for biosensing
New biomolecular recognition elements for analytical biotechnology (Luckenwalde, Germany)
1998年06月, 口頭発表（招待・特別） - DNA sensor
New biomolecular recognition elements for analytical biotechnology (Luckenwalde, Germany)
1998年06月, 口頭発表（招待・特別） - バイオセンサーと環境センシングの新展開
国際環境バイオ・エコ技術講演会 (tokyo, Japan)
1997年01月, 口頭発表（招待・特別）
外部研究資金等
- 国立精神・神経センター神経研究所金子清俊教授との共同研究 μmサイズの生体組織切片中に含まれるタンパク質の新規同定法の開発
共同研究, 自 0000年, 至 0000年 - DNAアプタマーに関する指導
受託研究, 自 2016年07月01日, 至 2017年03月31日 - 酵素阻害アプタマーを用いた高感度簡易迅速疾病診断法の開発
受託研究, 自 2016年04月01日, 至 2017年03月31日 - 酵素阻害アプタマーを用いた高感度簡易迅速疾病診断法の開発
受託研究, 自 2015年11月16日, 至 2016年03月31日 - アプタマーを用いた各種測定法の習得
受託研究, 自 2015年07月08日, 至 2015年07月09日 - 反応内臓チップによる小型遺伝子定量装置の実用化開発
受託研究, 自 2014年04月01日, 至 2015年03月31日 - 反応内臓チップによる小型遺伝子定量装置の実用化開発
受託研究, 自 2013年04月01日, 至 2014年03月31日 - 反応内臓チップによる小型遺伝子定量装置の実用化開発
受託研究, 自 2012年10月01日, 至 2013年03月31日 - 食品分析用バイオセンサの試作評価
受託研究, 自 2012年10月01日, 至 2013年03月31日 - マイクロ流路チップ・フローサイトメーターとアプタマーを用いたアルツハイマー病早期診断法の開発
受託研究, 自 2012年04月01日, 至 2012年09月30日 - アプタマーを利用した電気化学的VEGF高感度・迅速検出システムの開発
受託研究, 自 2012年04月01日, 至 2012年07月31日 - アプタマーを利用した電気化学的VEGF高感度・迅速検出システムの開発
受託研究, 自 2011年12月01日, 至 2012年03月31日 - マイクロ流路チップ・フローサイトメーターとアプタマーを用いたアルツハイマー病早期診断法の開発
受託研究, 自 2011年10月01日, 至 2012年03月31日 - 標的タンパク質認識アプタマーの分子進化
共同研究, 自 2010年04月01日, 至 2011年03月31日 - 平成20年度東京都重点戦略プロジェクト支援事業｢超高齢化都市のモデルに対応する有害微生物・ウイルスの複数・迅速・検出装置｣
共同研究, 自 2010年04月01日, 至 2011年02月28日 - 光熱変換とアプタマーを用いた標的タンパク質検出法の開発
共同研究, 自 2009年05月18日, 至 2010年03月31日 - 平成20年度東京都重点戦略プロジェクト支援事業 「超高齢化都市のモデルに対応する有害微生物・ウイルスの複数・迅速・検出装置」
共同研究, 自 2008年12月10日, 至 2009年03月31日 - 光熱変換とアプタマーを用いた標的タンパク質検出法の開発
共同研究, 自 2008年10月22日, 至 2009年03月31日 - 変形アプタマーによる酵素捕捉を利用した超高感度センシングシステムの開発
受託研究, 自 2008年06月17日, 至 2009年03月31日 - 臨床診断技術に関する研究
共同研究, 自 2008年05月22日, 至 2009年05月21日 - ペプチドリガンドの新規探索法in silico panning法の開発
受託研究, 自 2007年07月20日, 至 2008年03月31日 - レーザー干渉光熱変換法とアプタマーを用いた標的分子迅速検出技術の開発
受託研究, 自 2007年04月01日, 至 2007年10月31日 - 光導波路測定装置を用いたレジオネラ簡便検出用DNAセンシングシステムの開発
受託研究, 自 2006年, 至 2007年 - レーザ干渉光熱変換法とアプタマーを用いた標的分子迅速検出技術の開発
受託研究, 自 2006年, 至 2007年 - レーザー干渉光熱変換法とアプタマーを用いた標的分子迅速検出技術の開発
受託研究, 自 2006年11月01日, 至 2007年03月31日 - 干渉増幅反射法とアプタマーを利用した新規センシングシステムに関する研究
共同研究, 自 2006年01月17日, 至 2006年03月31日 - 標的タンパク質に結合するアプタマー材料の探索
共同研究, 自 2005年04月26日, 至 2006年03月31日 - リガンドの探索及び探索手法開発研究
共同研究, 自 2005年04月01日, 至 2006年03月31日 - 光導波路測定装置を用いた蛋白質の凝集・綿維化の解析
共同研究, 自 2005年04月01日, 至 2006年03月31日 - in silico 進化によるリガンドペプチドの探索
共同研究, 自 2005年02月10日, 至 2005年03月31日 - 生体分子設計法の研究開発
共同研究, 自 2004年, 至 2004年 - 標的タンパク質に結合するアプタマー材料の探索
共同研究, 自 2004年, 至 2004年 - 干渉増幅反射法を応用したバイオセンサーの開発
共同研究, 自 2004年10月18日, 至 2005年03月31日 - 標的タンパク質に結合するアプタマー材料の探索
共同研究, 自 2004年09月13日, 至 2005年03月31日 - 生体分子設計方法の開発研究
共同研究, 自 2004年06月22日, 至 2005年03月31日 - 光導波路蛍光測定装置とDNAアプタマーを用いた標的タンパク質検出システムの
共同研究, 自 2003年, 至 2004年 - 干渉増幅反射法による分子間相互作用を応用したバイオセンサーの構築
共同研究, 自 2003年, 至 2004年 - 微生物によるバイオ燃料電池変換技術の開発
受託研究, 自 2002年, 至 2004年 - 実用型SNP測定用チップの開発
受託研究, 自 1999年, 至 2000年 - 蛋白質の発現相互作用解析技術の開発
共同研究, 自 1999年, 至 2000年 - 環境測定用バイオセンサーの開発
共同研究, 自 1998年, 至 2000年
委員歴
メディア報道
- Researchers uncover novel amyloidosis α-S1-casein forms amyloid by overexpression and N-terminal truncation in canine mammary tumor
A collaboration led by scientists at Tokyo University of Agriculture and Technology (TUAT), Japan, has discovered a novel amyloid protein from canine mammary tumors.
EurekAlert!／News-Medical.Net／Phys.Org
自 2023年01月23日, 至 2023年01月24日 - 運転中の心筋梗塞、予兆を早期検知…デンソーなどDNAの機能を発見
デンソーと東京農工大学、理化学研究所、ノースカロライナ大学は、特殊な構造（グアニン四重鎖構造）を持つDNAがミオグロビンタンパク質の持つ酵素活性を増強する機能があることを発見したことが紹介される。
yahooニュース／Response／msnニュース
自 2021年06月08日, 至 2021年06月08日 - DNAに“設計図”以外の新機能を発見 酵素活性を強める効果
東京農工大学大学院が、DNAに「生命の設計図」以外の機能として、酵素活性を強める効果を見つけたと発表したことが紹介される。
yahooニュース／ITmedia／gooニュース／msnニュース／ニコニコニュース
自 2021年06月09日, 至 2021年06月09日 - 東京農工大など、ルミノール反応を300倍以上に増強するDNAアプタマーを開発
東京農工大学の池袋一典教授らの研究チームが、ミオグロビン蛋白質のルミノールに対する酸化反応の活性を300倍以上に増強するDNAアプタマーを開発したと発表したことが紹介される。
日経バイオテク
自 2021年06月11日, 至 2021年06月11日
Dr. Ikebukuro and his group have developed many DNA aptamers that bind proteins, especially enzymes.
自 2021年07月01日, 至 2021年07月01日- 新型コロナウイルスに対する安価、短時間、高感度の抗原検査法を開発
JNCと東京農工大学の池袋一典教授が、新型コロナウイルスの迅速‐高感度検出に成功したことが紹介される。
MONOist／yahooニュース
自 2020年12月14日, 至 2020年12月14日 - JNCなど 新型コロナ迅速検出 DNAアプタマー・検体混合 検出感度も向上
JNCと東京農工大学の池袋一典教授が、新型コロナウイルスの迅速‐高感度検出に成功したことが紹介される。
日刊工業新聞／化学工業日報／石油化学新聞日刊通信／日刊ケミカルニュース
自 2020年11月25日, 至 2020年11月25日 - JNC コロナを迅速かつ高感度に検出する技術を共同開発 JNC , 東京農工大学 , 迅速-高感度免疫診断技術AptⅠa（アプティア）法
JNCと東京農工大学の池袋一典教授が、新型コロナウイルスの迅速‐高感度検出に成功したことが紹介される。
日刊ケミカルニュース／Jchem-News
自 2020年11月25日, 至 2020年11月25日