マイクロペルオキシダーゼ類の完全化学合成とその構造・機能変換による酸化還元性人工タンパク質の開発とナノバイオセンサーへの応用
キーワード：直接電子移動反応，タンパク質，酵素，バイオセンサー
2015.10.
| 中野 幸二（なかの こうじ） | データ更新日：2024.04.19 |
主な研究テーマ
PNAzyme-ペプチド核酸酵素を基盤とするバイオ分析プラットホーム
キーワード：ペプチド核酸、コンジュゲート、バイオセンサ、バイオイメージング
2015.04.
キーワード：ペプチド核酸、コンジュゲート、バイオセンサ、バイオイメージング
2015.04.
光アップコンバージョン機能を組み込んだピロール・イミダゾール・ポリアミドの合成とDNA二重らせんのin vivoイメージング
キーワード：ピロール・イミダゾール・ポリアミド、光アップコンバージョン、バイオイメージング
2015.04.
キーワード：ピロール・イミダゾール・ポリアミド、光アップコンバージョン、バイオイメージング
2015.04.
酸化還元タグを組み込んだ新規核酸合成基質を用いる核酸の電気化学バイオアッセイの研究
キーワード：ペプチド核酸、コンジュゲート、バイオセンサ、バイオイメージング
2015.04.
キーワード：ペプチド核酸、コンジュゲート、バイオセンサ、バイオイメージング
2015.04.
化学修飾カーボンドットの作製とバイオセンシング・バイオイメージングへの応用
キーワード：カーボンドット、ナノクラスター、蛍光、バイオセンサー、バイオイメージング
2013.04.
キーワード：カーボンドット、ナノクラスター、蛍光、バイオセンサー、バイオイメージング
2013.04.
匂い分子感応性ポリペプチドホストの合成とバイオセンシング・バイオイメージングの研究
キーワード：バイオセンサー、バイオイメージング、ナノレポーター粒子、匂い
2011.10.
キーワード：バイオセンサー、バイオイメージング、ナノレポーター粒子、匂い
2011.10.
DNA超分子組織体を用いるナノバイオセンシング
キーワード：バイオセンサー、ナノバイオデバイス、DNA、超分子組織体
2000.10.
キーワード：バイオセンサー、ナノバイオデバイス、DNA、超分子組織体
2000.10.
原子間力顕微鏡を用いるバイオイメージングと１分子検出によるバイオ分析化学
キーワード：AFM、バイオイメージング、バイオ分析化学、１分子検出
2000.10.
キーワード：AFM、バイオイメージング、バイオ分析化学、１分子検出
2000.10.
DNAバイオセンサー・遺伝子センサー
キーワード：DNA，遺伝子，バイオセンサー，走査電気化学顕微鏡，ハイスループットアッセイ
1992.04～2020.03.
キーワード：DNA，遺伝子，バイオセンサー，走査電気化学顕微鏡，ハイスループットアッセイ
1992.04～2020.03.
タンパク質・酵素の直接電子移動反応とバイオセンサー応用
キーワード：直接電子移動反応，タンパク質，酵素，バイオセンサー
1995.10～2020.03.
キーワード：直接電子移動反応，タンパク質，酵素，バイオセンサー
1995.10～2020.03.
超分子組織体を感応素子に用いる化学計測法
キーワード：超分子組織体，合成二分子膜，自己組織化単分子膜，多相系高分子材料
1985.04.
キーワード：超分子組織体，合成二分子膜，自己組織化単分子膜，多相系高分子材料
1985.04.
従事しているプロジェクト研究
世界初のDX型研究方に基づく新しい産学連携・教育システム・オープンサイエンスプラットホーム
2020.10～2029.09, 代表者：片山佳樹, 九州大学大学院工学研究院, 日本.
2020.10～2029.09, 代表者：片山佳樹, 九州大学大学院工学研究院, 日本.
九州大学大学院博士課程教育リーディングプログラム「分子システムデバイスコース」
2012.10～2018.03, 代表者：安達千波矢, 九州大学大学院工学研究院, 日本.
2012.10～2018.03, 代表者：安達千波矢, 九州大学大学院工学研究院, 日本.
九州大学「先端分子技術を核とする九州先端ものづくりセンターの構築（クリーン実験ステーション）」
2014.04～2017.03, 代表者：浅野種正, 九州大学大学院システム情報研究院, 日本
文部科学省「先端研究施設共用促進事業」に採択された「九州大学 先端分子技術を核とする九州先端ものづくりセンターの構築（クリーン実験ステーション）」の運営委員として活動した。.
2014.04～2017.03, 代表者：浅野種正, 九州大学大学院システム情報研究院, 日本
文部科学省「先端研究施設共用促進事業」に採択された「九州大学 先端分子技術を核とする九州先端ものづくりセンターの構築（クリーン実験ステーション）」の運営委員として活動した。.
平成23年度戦略的情報通信研究開発推進制度「匂いイメージセンサによる情報抽出に関する研究開発」
2011.10～2013.09, 代表者：林 健司, 九州大学大学院システム情報研究院, 日本
平成23年度戦略的情報通信研究開発推進制度に採択された「匂いイメージセンサによる情報抽出に関する研究開発」において、蛍光性ナノ粒子、化学受容性ペプチドを用いる匂い分子ナノレポーター粒子の研究を行。.
2011.10～2013.09, 代表者：林 健司, 九州大学大学院システム情報研究院, 日本
平成23年度戦略的情報通信研究開発推進制度に採択された「匂いイメージセンサによる情報抽出に関する研究開発」において、蛍光性ナノ粒子、化学受容性ペプチドを用いる匂い分子ナノレポーター粒子の研究を行。.
九州大学「先端分子技術を核とする九州先端ものづくりセンターの構築（クリーン実験ステーション）」
2009.04～2013.03, 代表者：浅野種正, 九州大学大学院システム情報研究院, 日本
文部科学省「先端研究施設共用促進事業」に採択された「九州大学 先端分子技術を核とする九州先端ものづくりセンターの構築（クリーン実験ステーション）」の運営委員として活動した。.
2009.04～2013.03, 代表者：浅野種正, 九州大学大学院システム情報研究院, 日本
文部科学省「先端研究施設共用促進事業」に採択された「九州大学 先端分子技術を核とする九州先端ものづくりセンターの構築（クリーン実験ステーション）」の運営委員として活動した。.
九州大学「未来分子システム科学」
2007.10～2012.03, 代表者：君塚信夫, 九州大学大学院工学研究院, 日本.
2007.10～2012.03, 代表者：君塚信夫, 九州大学大学院工学研究院, 日本.
九州大学「先端融合医療レドックスナビ研究連合拠点」
2007.05, 代表者：内海英雄, 九州大学大学院薬学研究院, 日本.
2007.05, 代表者：内海英雄, 九州大学大学院薬学研究院, 日本.
九州大学「先端融合医療レドックスナビ研究拠点」（ＦＳ）
2006.04～2007.03, 代表者：内海英雄, 九州大学, 文部科学省科学技術振興調整費「先端融合領域イノベーション創出拠点の形成」
我々生命体はその活動を様々なレドックス反応を基本として維持しているが、近年、種々の環境要因の変動により、あるいは非健康的な生活習慣によりこのレドックス代謝に異常を起こし、がん、生活習慣病、脳神経変性疾患等を引き起こすことが明らかになってきた。本プロジェクト研究では、生体レドックスを詳細に評価し、自在に操ることのできる統合技術、具体的には早期診断・治療法、治療薬の開発をめざす。.
2006.04～2007.03, 代表者：内海英雄, 九州大学, 文部科学省科学技術振興調整費「先端融合領域イノベーション創出拠点の形成」
我々生命体はその活動を様々なレドックス反応を基本として維持しているが、近年、種々の環境要因の変動により、あるいは非健康的な生活習慣によりこのレドックス代謝に異常を起こし、がん、生活習慣病、脳神経変性疾患等を引き起こすことが明らかになってきた。本プロジェクト研究では、生体レドックスを詳細に評価し、自在に操ることのできる統合技術、具体的には早期診断・治療法、治療薬の開発をめざす。.
21世紀COEプログラム九州大学「分子情報科学の機能イノベーション」
2002.10～2007.03, 代表者：新海征治, 九州大学, 文部科学省（日本）
文部科学省が平成14年度から開始した、「我が国の大学に世界最高水準の研究教育拠点を学問分野毎に形成し、研究水準の向上と世界をリードする創造的な人材育成を図るため，重点的な支援を行い，国際競争力のある個性輝く大学づくりを推進すること」拠点形成事業である。当該の拠点形成事業では、分子を基盤とする「モレキュラーインフォーマティクス（分子情報科学）」の研究分野を構築するとともに、1) 院生プロジェクト推進、2) 国際化教育、3) 産学連携教育の３本を柱として、「ボーダーレス時代」といわれる21世紀に世界を舞台に飛躍できる人材の育成をめざす。.
2002.10～2007.03, 代表者：新海征治, 九州大学, 文部科学省（日本）
文部科学省が平成14年度から開始した、「我が国の大学に世界最高水準の研究教育拠点を学問分野毎に形成し、研究水準の向上と世界をリードする創造的な人材育成を図るため，重点的な支援を行い，国際競争力のある個性輝く大学づくりを推進すること」拠点形成事業である。当該の拠点形成事業では、分子を基盤とする「モレキュラーインフォーマティクス（分子情報科学）」の研究分野を構築するとともに、1) 院生プロジェクト推進、2) 国際化教育、3) 産学連携教育の３本を柱として、「ボーダーレス時代」といわれる21世紀に世界を舞台に飛躍できる人材の育成をめざす。.
１分子検出のための遺伝子プローブ−ＤＮＡリガンド−の研究
2005.10～2006.03, 科学技術振興機構（日本）
特定遺伝子の１分子検出・分析法の研究を行う。この目的に、新規ＤＮＡプローブ−ＤＮＡリガンド−の開発研究を行い。.
2005.10～2006.03, 科学技術振興機構（日本）
特定遺伝子の１分子検出・分析法の研究を行う。この目的に、新規ＤＮＡプローブ−ＤＮＡリガンド−の開発研究を行い。.
DNA二重らせんを電子機能・構造単位とする単一分子素子
2000.10～2003.09, 科学技術振興事業団「さきがけ」（日本）
DNA二重らせんと低分子化合物からのコンジュゲート形成を利用して合目的な機能付与を行い、電子材料への応用と単一分子素子への展開を図る。.
2000.10～2003.09, 科学技術振興事業団「さきがけ」（日本）
DNA二重らせんと低分子化合物からのコンジュゲート形成を利用して合目的な機能付与を行い、電子材料への応用と単一分子素子への展開を図る。.
研究業績
主要著書
主要原著論文
| 1. | T. Anada, H. Matsunaga, R. Karinaga, K. Koumoto, M. Mizu, K. Nakano, S. Shinkai, and K. Sakuraia, Proposal of new modification technique for linear double-stranded DNAs using the polysaccharide schizopyllan, Bioorganic & Medicinal Chemistry Letters, 10.1016/j.bmcl.2004.08.043, 14, 22, 5655-5659, Vol.14, pp.5655 - 5659, 2004.09, A natural polysaccharide schizophyllan (SPG) has been known to form a stable complex with poly(dA). We attached a poly(dA)(80) tail to the both ends of a linear double-stranded DNA, which had been prepared from a plasmid DNA vector. The poly(dA) tailed DNA verified to form complex with SPG by gel electrophoresis and atomic force microscopy (AFM). AFM images indicated that the complexes exhibit a dumbbell-like architecture, that is, quite similar to that of adenovirus genome. The complex demonstrated excellent exonuclease resistance, probably because of the protection effect by SPG complexation.. |
| 2. | K. Nakano, T. Sawada, Y. Mori, K. Morita, R. Ishimatsu, Covalent Hyperbranched Polymer Self-Assemblies of Three-Way Junction DNA for Single-Molecule Devices, Langmuir, https://dx.doi.org/10.1021/acs.langmuir.0c01621, 36, 34, 10166-10174, 2020.09, A hyperbranched polymer (HBP) made of three-way junction (TWJ) DNAs is reported. Three types of 26-mer DNAs with 5′-ends modified with psoralen (PSN) were synthesized. All had self-complementary sequences starting from the 5′-end to the sixth base (AAGCTT), allowing intermolecular hybridization. The base sequences of the remaining 20-mer sites were designed so that upon hybridization, three strands had a TWJ structure with a mass of 25,000 that could be further grown by forming HBPs. PSN photochemically reacts to form interstrand cross-links that increase the polymer stability. Aggregates [(380 ± 44) nm and (65 ± 6) nm] detected with dynamic light scattering for TWJ-DNA solutions were also imaged by electron microscopy and atomic force microscopy, providing evidence of hyperbranched polymerization. The TWJ unit also polymerized on solid substrates such as Au and glass and formed self-assembled monolayers (SAMs). The HBP SAMs were integrated into commercial Pt-interdigitated electrode arrays. The DNA devices had current–voltage curves typical of metal–insulator–metal Schottky diodes; the effective barrier heights and the ideality factors were 0.52 ± 0.002 eV and 21 ± 3.2, respectively. The series resistances were (26 ± 3.3) × 10^6 Ω, which may provide insights into DNA electron transport. The DNA HBP enables stable electrical connections with probe electrodes and will be an important single-molecule platform.. |
| 3. | K. Nakano, J. Horiuchi, S. Hirata, M. Yamanaka, H. Himeno, R. Ishimatsu, Folding and Assembly of Vanilloid Receptor Secondary-Structure Peptide with Hexahistidine Linker at Nickel−Nitrilotriacetic Acid Monolayer for Capsaicin Recognition, Langmuir, DOI: 10.1021/acs.langmuir.8b03202, 35, 6, 2047-2054, 2019.02, Herein, we report the self-assembly of a synthetic vanilloid receptor (VR) peptide that selectively binds capsaicin. We synthesized a 26-mer peptide—YSEILFFVQS-HHHHHH-LAMGWTNMLY (S3HS4)—comprising two chemoreceptor domains of transient receptor potential channel (TRPV1) linked by a hexahistidine sequence. High-speed atomic force microscopy (AFM) imaging in water revealed that the peptide structures alternated rapidly between wedge shape and linear forms. Circular dichroism spectroscopy showed that 65% of the amide units in the peptide chain adopted an α-helix structure, which was ascribed to the chemoreceptor domains. S3HS4 developed well-packed monolayers at the Ni-treated thiolated nitrilotriacetic acid self-assembled monolayers by chelation of the hexahistidine segment, as characterized by infrared spectroscopy and AFM, which exhibited statistically constant specific height. Therefore, S3HS4 was expected to fold spontaneously upon chelation, and the resulting helix−turn−helix conformers developed films while uniformly oriented: the tilt angle was 69° from the surface normal to the substrate. According to microgravimetric analysis using a quartz crystal microbalance (QCM), the adsorption was 84 ± 47 pmol cm−2 (n = 3), which was almost consistent with the saturation adsorption of an α-helix unit. We also used a QCM to investigate the host−guest reactions of S3HS4 and found that the S3HS4-attached QCM-chip-bound capsaicin with an apparent binding constant of (4.2 ± 3.6) × 104 M−1 (n = 4), whereas there was no evidence of binding to vanillin or acetophenone. Two controls—a blank chip without S3HS4 and a chip modified with a single helical peptide (LAMGWTNMLY-HHHHHH)—produced no capsaicin response. To the best of our knowledge, S3HS4 is the first example of a synthetic VR mimic peptide. We believe that the present surface-directed structure-based design can be used to exploit the α-helix bundle in hexahistidine-linked bishelical peptides.. |
| 4. | Koji Nakano, Takayuki Honda, Kanako Yamasaki, Yoshiki Tanaka, Keiichi Taniguchi, Ryoichi Ishimatsu, Toshihiko Imato, Carbon quantum dots as fluorescent component in peroxyoxalate chemiluminescence for hydrogen peroxide determination, Bulletin of the Chemical Society of Japan, 10.1246/bcsj.20180095, 91, 7, 1128-1130, 2018.01, [URL], Nitrogen-doped carbon quantum dots synthesized by onepot, microwave-assisted pyrolysis of citric acid in the presence of a small number of N-doping precursors, 1, 2-ethylenediamine, were found to be involved in the chemically initiated electron exchange luminescence enabling peroxyoxalate chemiluminescence assay of hydrogen peroxide in the concentration range of 101000 μM.. |
| 5. | J. Tanabe, K. Nakano, R. Ishimatsu, T. IMATO, H. Okabe, N. Matsuda, Totally Synthetic Microperoxidase-11, Royal Society Open Science, http://dx.doi.org/10.1098/rsos.172311, 5, 3, 172311-172320, 2018.05, A totally synthetic microperoxidase-11 (MP-11) is reported. Accordingly, the undecapeptide (VQKCAQCHTVE) was synthesized by solid-phase peptide synthesis followed by the thiol-ene click reaction with haemin for reconstitution. High-speed atomic force microscopy measurement conducted in water confirmed the protein reconstitution by visualizing the morphological differences as animated molecular images. The synthetic MP-11 showed a considerable magnitude of catalytic activity (27%) against the natural MP-11 in the oxidation of 3,3′,5,5′-tetramethylbenzidine by hydrogen peroxide, whereas it showed very low (2.7%) activity of a synthetic variant with a point mutation (VQKCAQCMTVE, H8M). Slab waveguide spectroscopic measurements revealed that the ferrous/ferric redox reaction occurred by the direct electron transfer with specific spectral changes. Indeed, if hydrogen peroxide existed in the solution phase, the peroxidase-modified electrode showed catalytic current–voltage behaviour regardless of whether it was prepared using natural MP-11 or the synthetic MP-11. If a substrate recycling reaction was assumed, computer simulation well reproduced the experimental curves to give a global set of electrocatalytic reaction parameters. In any of the experiments, the synthetic MP-11 and natural MP-11 gave almost identical results. Our approach will be a convenient means of preparing MP-11, as well as its mutants, that does not rely on nature.. |
| 6. | K. Nakano, J. Tanabe, R. Ishimatsu, T. IMATO, Monolithic Peptide-Nucleic Acid Hybrid Functioning as an Artificial Microperoxidase, Bioconjugate Chemistry, https://doi.org/10.1021/acs.bioconjchem.7b00216, 28, 5, 2031-2034, 2017.05, A new peptide nucleic acid (PNA) with an installed peroxidase function has been developed. Fmoc solid phase peptide synthesis prepared a PNA hy-brid (VQKCAQCHTVE-(C2H4O)2CH2-[PNA(T)]6-G) that renders the microperoxidase backbone, fol-lowed by reconstitution with hemin. The resulting holocompound catalyzed the oxidation of 3,3',5,5'-tetramthylbenzidine by H2O2 to 50% that of natural microperoxidase-11, whereas the apo-form and hemin gave no responses. The peroxidase domain was found to be active toward direct electrochemis-try and the PNA hybrid served for gene sensor; in the presence of the target DNA (5'-CATGTATAAAAAA-3'), an electrode-attached DNA probe (5'-TsTsTsTsTsTCTCATACATG-3') showed the ferric-to-ferrous quasi-reversible wave (–276 mV vs. Ag/AgCl) through sandwich hybridi-zation. Moreover, the hybridization product could accept H2O2 as an oxidant to enhance the reduction current, which occurred likely based on the iron(II)-center-recycling with specific rate constant of 0.19 s–1.. |
| 7. | Koji Nakano, Shingo Hirata, Jun Horiuchi, Ryoichi Ishimatsu, Toshihiko Imato, Takeshi Onodera, Kenshi Hayashi, Synthesis and Self-Assembly of His-tag Hybrid of Substrate-Binging Short Domain in Transient Receptor Potential Vanilloid Type 1 for Vanaillin Sensing Application, Transactions of the Materials Research Society of Japan, http://doi.org/10.14723/tmrsj.40.175, 40, 2, 175-178, 2015.07. |
| 8. | Koji Nakano, Takayuki Kimura, Yosuke Kitamura, Toshihiro Ihara, Ryoichi Ishimatsu, Toshihiko Imato, Potentiometric DNA Sensing Platform Using Redox-active DNA Probe Pair for Sandwich-type Dual Hybridization at Indicator Electrode Surface, Journal of Electroanalytical Chemistry, http://dx.doi.org/10.1016/j.jelechem.2014.03.029, 720-721, 71-75, 2014.04, A potentiometric sensing platform that enables real-time DNA hybridization detection is described. A model target DNA, t37s: 5'AAA AAA AAA AAA-(TC)2-Ts5-(TC)2-GGA GCT GGT GGC3', which consist of a dodecamer polydeoxyadenylic acid and the human K-ras oncogene, and a five-successive deoxythymidine phosphorothioate (Ts) was designed. With the gold–phosphorothioate binding, the DNA could bind the complementarily sequences to concentrate them at the gold electrode surfaces. Accordingly, a dodecamer polydeoxythymidylic acid having ferroin-moiety (T12FeP) and a ferrocene-modified complementary of K-ras (KrasFc) were synthesized. Electrochemical quartz-crystal-microbalance (QCM) using Au-sputtered quartz chips that served as the indicator electrode collected the electrode responses. When the electrode surface was treated with the T12FeP-hybridizad t37s, which were subsequently oxidized to the corresponding Fe(III) form, the emf developed in a buffer solution responded to KrasFc; with the electrode-attached t37s the redox-active DNAs could group into a pair to establish specific redox-titration equilibrium. A QCM confirmed the on-electrode titrimetry feasible by determining the initial concentration and the amount of hybridization along with the emf measurements. Although the present method is necessary for two-kinds of redox conjugation of the target DNA, it should be important as a novel framework of electrochemical gene sensing. Preliminary examples of real-time measurement were also demonstrated with the results of kinetics analysis data.. |
| 9. | 中野 幸二, 澤田 貴文, 森 美詞, 石松 亮一, 今任 稔彦, 光架橋性オリゴヌクレオチドからのDNAデンドリマー自己組織化膜形成, 電子情報通信学会技術研究報告, 112, 456, 29-32, 2013.03, We designed three types of 26-mer oligodeoxyribonucleotides that produce the three-way junction (TWJ) structure through the selective hydrogen bonding between the complimentary base-pairs. At the 5'-end was left a self-complimentary sticky sequence, which was further modified with psoralen (7H-furo[3,2-g]chromen-7-one)to exert covalent molecular aggregation through the interstrand photochemical crosslinking. Dynamic light scattering as well as high-resolution electron microscopy revealed that the TWJ unit developed very large aggregates in solution, a new kind of dendrimer made by the TWJ-DNA duplex, which were ca. 400-nm in diameter having three-dimensional network structure. We further examined the TWJ-DNA molecular aggregation at the solid substrates. Interestingly, we found that the TWJ-DNA duplexes self-assembled to developed uniform dendrimer monolayers. Various surface analysis methods including cyclic voltammetry and electrochemical impedance spectroscopy characterized the DNA dendrimer SAMs in detail.. |
| 10. | Hisao Yoshinaga, Koji Nakano, Nobuaki Soh, Ishimatsu Ryoichi, Toshihiko Imato, A Pivot-Hinge-Style DNA Immobilization Method with Adaptable Surface Concentration Based on Oligodeoxynucleotide-Phosphorothioate Chemisorption on Gold Surfaces, Anal. Sci., https://doi.org/10.2116/analsci.28.1059, 28, 11, 1059-1064, 2012.11. |
| 11. | Hisao Yoshinaga, Koji Nakano, Nobuaki Soh, Toshihiko Imato, AFM-Imaging Diagonosis Method for Single Nucleotide Polymorphism Using Molecular Beacon DNA as an Intramolecullar Ligation Template of Target DNA and a Viewable Indicator, Anal. Sci., https://doi.org/10.2116/analsci.28.939, 28, 10, 939-945, 2012.10. |
| 12. | 北岡 桃子, Masayuki Mitsumori, Kounosuke Hayashi, Yoshiyuki Hiraishi, Hisao Yoshinaga, Koji Nakano, Katsuyuki Miyawaki, Sumihare Noji, Masahiro Goto, Noriho Kamiya, Transglutaminase-Mediated in Situ Hybridization (TransISH) System: A New Methodology for Simplified mRNA Detection, Anal. Chem., https://doi.org/10.1021/ac2034198, 84, 14, 5885-5891, 2012.07. |
| 13. | Josui Shimada, Tatsuo Maruyama, 北岡 桃子, Hisao Yoshinaga, Koji Nakano, Noriho Kamiya, Masahiro Goto, Programmable protein–protein conjugation via DNA-based self-assembly, Chem. Commun., https://doi.org/10.1039/C2CC30618B, 48, 50, 6226-6228, 2012.06. |
| 14. | Koji Nakano, Hirokazu Yamanouchi, Hisao Yoshinaga, Nobuaki Soh, Toshihiko Imato, Label-free DNA Detection Platform Based on Atomic Force Microscopy Visualisation: Characterising the Molecular-Recognition-Triggered Conformational Changes of an Immobilised Receptor Oligonucleotide Probe, Chem. Commun., https://doi.org/10.1039/C002642E, 46, 13, 5683-5685, 2010.06, Using atomic microscopy imaging, probe DNA sequence self-assemblies developed on Si(100) substrates undergo a conformational transition from an extended stem-loop structure to a double helix; such assemblies readily report on DNA molecular recognition events and should be suitable as a label-free, DNA hybridisation assay platform.. |
| 15. | Koji Nakano, Yosuke Katsumi, Nobuaki Soh, Toshihiko Imato, An Atomic Force Microscopy Assay of Intercalation Binding, Unwinding and Elongation of DNA, Using a Water-Soluble Psoralen Derivative as a Covalent Binding Probe Molecule, Bull. Chem. Soc. Jpn, https://doi.org/10.1246/bcsj.20090166, 2010, 3, 273-275, 2010.03, A single-molecule strategy using atomic force microscopy has simply yet robustly probed an intercalation binding related specific structural relaxation of covalently closed circular pBR322 DNA as well as double-strand elongation in its linear form by taking advantage of a new psoralen derivative, N,N,N-trimethyl-1-(2,5,9-trimethyl-7-oxo-7Hfuro[3,2-g]chromen-3-yl)methanaminium chloride, that covalently binds to DNA through a photo-crosslinking reaction.. |
| 16. | 中野幸二，吉武忠輝，山下泰徳，宗 伸明，今任稔彦, アルカンチオール単分子膜修飾電極に固定化したチトクロームcの直接電子移動反応の研究, 九州大学中央分析センター報告, 27, 17-24, 2010.12. |
| 17. | Koji NAKANO, Hideshi MATSUNAGA, Masaharu MURATA, Nobuaki SOH, Toshihiko IMATO, Synthesis Of Circular Double-Stranded DNA Having Single-Stranded Recognition Sequence As Molecular-Physical Probe For Nucleic Acid Hybridization Detection Based On Atomic Force Microscopy Imaging, Anal. Sci., DOI: 10.2116/analsci.25.993, 25, 8, 993-998, 2009.08, A new class of DNA probes having a mechanically detectable tag is reported. The DNA probe, which consists of a single-stranded recognition sequence and a double-stranded circular DNA entity, was prepared by polymerase reaction. M13mp18 single strand and a 32mer oligodeoxynucleotide whose 5`-end is decorated with the recognition sequence were used in combination as template and primer, respectively. We have successfully demonstrated that the DNA probe is useful for bioanalytical purposes: by deliberately attaching target DNA molecules onto Au(111) substrates and by mechanically reading out the tag-entity using a high-resolution microscopy including atomic force microscopy, visualization/detection of the individual target/probe DNA conjugate was possible simply yet straightforwardly. The present DNA probe can be characterized as a 100%-nucleic acid product material. It is simply available by one-pod synthesis. A surface topology parameter, image roughness, has witnessed its importance as a quantitative analysis index with particular usability in the present visualization/detection method.. |
| 18. | K. Nakano, K. Nakamura, K. Iwamoto, N. Soh, T. Imato, Positive-feedback-mode scanning electrochemical microscopy imaging of redox-active DNA-poly(1,4-benzoquinone) conjugate deposited on carbon electrode for micrometer-sized hybridization biosensor applications, J. Electroanal. Chem., https://doi.org/10.1016/j.jelechem.2009.01.014, 628, 1-2, 113-118, Vol. 628, No. 1, pp. 113–118, 2009.02, Scanning electrochemical microscopy of DNA microdots deposited on carbon fiber microelectrodes (diameter, 33 μm) has been demonstrated. The microdots, which comprised quinone polymer matrices with capture probe (CP) DNA grafted onto them, can report hybridization events via changes in their electrochemical reactivity. Furthermore, the polymer matrices, even after conjugation with CP DNA, possess a certain degree of charge-transport capability and thus allow for positive-feedback-mode imaging. We have successfully obtained well-resolved micrometer-sized dot images (diameter, 60–100 μm) of the microelectrodes: they generate a considerable magnitude of current rise over 10 nA while they gave a current decrease, typically 1 nA, in responding the hybridization event at the CP DNA. The sensor response was found to fall a little larger than the background current (0.6–0.8 nA). However, the particular SECM measurement system represented good signal-to-noise ratio reliably allowing the detection of DNA hybridization feasible. Obtaining these results, we have concluded that the particular DNA-modified electrode with SECM detection should be useful for readout of DNA hybridization sensor coupled with a high-throughput-device such as DNA microarrays.. |
| 19. | K. Nakano, K. Ohkubo, H. Taira, M. Takagi, N. Soh, T. Imato, Synthesis of 1,4-hydroquinone-terminated alkanethiol and self-assembly on gold as characterized by interfacial electrochemistry, electrocatalysis application and ab initio calculation based on comparison with catechol-presenting analogue., J. Electroanal. Chem., https://doi.org/10.1016/j.jelechem.2008.06.016, 623, 1, 49-53, Vol. 623, No. 1, pp. 49-53, 2008.11, Synthesis and self-assembly of a mercaptoundecaneamide derivative having a terminus of 1,4-hydroquinone (QT) are described. Electrochemical measurements on the QT-modified Au electrode revealed that the alkanethiol compound undergoes self-assembly to exhibit specific electrochemical activity originates from the reversible quinone/hydroquinone redox reaction at the terminus. We have achieved to obtain the electrochemical active surface coverage (0.11 nmol cm−2), formal potential (+246 mV, pH 3, Ag/AgCl) that changes pH-dependently (58 mV per pH) and also a set of the electron transfer reaction parameters, all of which were consistent with those of the previously reported structural isomer, catechol-terminated mercaptoundecaneamide (CT). Contrastingly, we found that these alkanethiol monolayers give marked contrast in an elecrocatalysis application: the CT-monolayer electrodes showed electrocatalytic capability in oxidation of NADH solution species while the QT-monolayer electrodes did not at all. By comparing some results of theoretical approach, we have attributed the surface selectivity to the spatiality of particular molecular orbital in the catalysis molecule. This observation should be important as an example of spatiality–reactivity relationships in a molecular design of chemically modified electrode.. |
| 20. | K. Nakano, K. Ohkubo, H. Taira, M. Takagi, T. Imato, Electrocatalytic oxidation of dihydronicotineamide adenine dinucleotide on gold electrode modified with catechol-terminated alkanethiol self-assembly, Anal. Chim. Acta, https://doi.org/10.1016/j.aca.2008.02.009, 619, 1, 30-36, Vol. 619, No. 1, pp. 30–36, 2008.06, Synthesis of a mercaptoundecaneamide derivative having a terminus of catechol is described. FT-IR spectroscopic characterization showed that the new molecular entry simply undergoes molecular self-assembly on Au substrate surfaces promoting intra- and intermolecular hydrogen bonds to form well-packed monolayers. Cyclic voltammetric (CV) measurements on the monolayer-modified Au electrode revealed that the surface adlayer possesses specific electrochemical activity due to the reversible catechol/o-quinone redox reaction having characteristics of a surface process and also pH-dependence in its formal potential (59 mV per pH). Detailed analysis of CVs gave fundamental electrochemical parameters including the electroactive surface coverage (0.20–0.24 nmol cm−2), the transfer coefficients (0.24 in oxidation and 0.81 in reduction), and also the electron transfer rate constant (1.10–2.76 s−1). These data were almost consistent to those seen in literature. We have also found that the catechol monolayer modified electrode exhibits an electrocatalytic function in NADH oxidation. That is, the faradaic current appeared reinforcingly at around the same potential where catechol function is oxidized in the monolayer and increased with an increase in the NADH concentration from 1 to 5 mM, and then reached to a plateau indicating a catalyzed reaction pathway. Detailed analyses revealed that the present system could be characterized by its weak stability of the intermediate compound formed and prompt reaction rate compared with the previously reported chemically modified electrode (CME) systems. We think this type of achievement should be important for the basics of biosensors that rely on dehydrogenase enzymes.. |
| 21. | N. Soh, K. Makihara, T. Ariyoshi, D. Seto, T. Maki, H. Nakajima, K. Nakano, T. Imato, Phospholipid-linked Coumarin: A Fluorescent Probe for Sensing Hydroxyl Radicals in Lipid Membrane, Analytica Sciences, https://doi.org/10.2116/analsci.24.293, 24, 2, 293-296, Vol. 24, No. 2, pp. 293–296, 2008.02, A fluorescent probe, DPPEC (1,2-dipalmitoylglycerophosphorylethanolamine labeled with coumarin) was developed for detecting hydroxyl radical (·OH) in lipid membranes. The coumarin moiety contributes to the fluorescent detection of ·OH and the phospholipids moiety gives a driving force to localize the probe in lipid membranes. DPPEC in liposomal membranes rapidly reacted with ·OH and increased the fluorescence intensity, depending on the concentration of ·OH. The increase in the fluorescence intensity induced by ·OH was effectively suppressed by the addition of DMSO. The probe exhibited a higher fluorescence response to ·OH over other reactive oxygen species, such as hydrogen peroxide, nitric oxide, peroxynitrite, alkylperoxyl radical, and hypochlorite. DPPEC would be useful as a new type of fluorescent probe that can localize in lipid membranes and detect ·OH efficiently.. |
| 22. | N. Soh, K. Yoshida, H. Nakajima, K. Nakano, T. Imato, T. Fukaminato, M. Irie, A fluorescent photochromic compound for labeling biomolecules, Chemical Communications, DOI https://doi.org/10.1039/B713663C, 2007, 28, 5206-5208, Vol. 2007, No. 28, pp. 5206–5208, 2007.12, A fluorescent photochromic compound, composed of diarylethene, fluorescein and succinimidyl ester units, was developed for the controllable fluorescent labeling of biomolecules based on a small molecule.. |
| 23. | N. Soh, T. Ariyoshi, T. Fukaminato, H. Nakajima, K. Nakano, T. Imato, Swallow-tailed perylene derivative: a new tool for fluorescent imaging of lipid hydroperoxides, Organic Bimolecular Chemistry, https://doi.org/10.1039/B713223A, 5, 23, 3762-3768, Vol. 5, No. 23, pp. 3762–3768, 2007.12, A swallow-tailed perylene derivative including a triphenylphosphine moiety was synthesized and applied to the detection and the live-cell imaging of lipid hydroperoxides. The novel probe, named Spy-LHP, reacted rapidly and quantitatively with lipid hydroperoxides to form the corresponding oxide, Spy-LHPOx, which emits extremely strong fluorescence (Φ ∼ 1) in the visible range (λem = 535 nm, 574 nm). Spy-LHP was highly selective for lipid hydroperoxides, and the addition of other reactive oxygen species (ROS) including hydrogen peroxides, hydroxyl radical, superoxide anion, nitric oxide, peroxynitrite, and alkylperoxyl radical, caused no significant increase in the fluorescence intensity. The probe exhibited good localization to cellular membranes and was successfully applied to the confocal laser scanning microscopy (CLSM) imaging of lipid hydroperoxides in live J774A.1 cells, in which lipid peroxidation was proceeded by the stimulation of 2,2-azobis(2-amidinopropane)dihydrochloride (AAPH). These findings establish Spy-LHP as a promising new tool for investigating the physiology of lipid hydroperoxides. . |
| 24. | 田中真由美，阪本一平，中嶋 秀，宗 伸明，中野幸二，Duck-Hwa CHUNG，今任稔彦, メチルパラチオン計測のための表面プラズモン共鳴イムノセンサーの開発, 分析化学, https://doi.org/10.2116/bunsekikagaku.56.705, 56, 9, 705-712, Vol. 56, No. 23, pp. 705–712, 2007.09, 表面プラズモン共鳴現象（SPR）を利用したオンサイトで簡便に残留農薬を計測できるフロー型イムノセンサーの開発を目的とし，メチルパラチオン（MP）の間接競合イムノアッセイ法を確立した．牛血清アルブミン（BSA）にMPを化学結合したMP-BSAコンジュゲートをSPRセンサーチップ上に物理吸着によって固定化し，これをフローシステムに組み込み，MPに対する応答性を検討した．SPRセンサーチップは，50 ppm MP-BSAコンジュゲート溶液をセンサーチップへ導入し，続いて10 mg/mL BSA溶液を導入し，非特異吸着を防ぐために未修飾のチップ表面をブロッキングして作製した．MP-BSAコンジュゲートのセンサーチップへの固定化密度を，飽和吸着量の約20% として作製したセンサーチップを用いて，間接競合法により60 ppm抗MP抗体を含むMP溶液（1～5000 ppb）をSPRセンサーに導入し，SPRセンサーの共鳴角度変化を測定した．本法によるMPの検出限界は，MP濃度ゼロにおける角度変化の85% におけるMP濃度とすると，10 ppbであった．また，ペプシンを含むpH 2 HCl-glycine溶液をセンサーチップに導入することによって，センサーチップ上のMP-BSAコンジュゲートと結合した抗MP抗体を解離させることができ，1枚のセンサーチップを用いて少なくとも20回繰り返し測定が可能であることが分かった．. |
| 25. | RuiQi Zhang, Hizuru Nakajima, Nobuaki Soh, Koji Nakano, Takashi Masadome, Kazumi Nagata, Kazuhira Sakamoto, Toshihiko Imato, Sequential Injection Chemiluminescence Immunoassay for Nonionic Surfactants Using Magnetic Microbeads, Analytica Chimica Acta, DOI: 10.1016/j.aca.2007.02.052, 600, 1-2, 105-113, Vol. 600, No. 1, pp. 105–113, 2007.09, A rapid and sensitive immunoassay based on a sequential injection analysis (SIA) using magnetic microbeads for the determination of alkylphenol polyethoxylates (APnEOs) is described. An SIA system was constructed from a syringe pump, a switching valve, a flow-through type immunoreaction cell equipped with a photon counting unit and a neodymium magnet. Magnetic beads, to which an anti-APnEOs monoclonal antibody was immobilized, were used as a solid support in an immunoassay. The introduction, trapping and release of the magnetic beads in and from the immunoreaction cell were controlled by means of a neodymium magnet and adjusting the flow of a carrier solution. The immunoassay was based on an indirect competitive immunoreaction of an anti-APnEOs monoclonal antibody immobilized on the magnetic beads with a sample APnEOs and a horseradish peroxidase (HRP)-labeled APnEOs in the same sample solution, and was based on the subsequent chemiluminscence reaction of HRP on the magnetic microbeads with a luminol solution containing hydrogen peroxide and p-iodophenol. The anti-APnEOs antibody was immobilized on the magnetic microbeads by coupling the antibody with the magnetic beads after activation of a carboxylate moiety on the surface of the magnetic beads that had been coated with a polylactic acid film. The antibody immobilized magnetic beads were introduced in the immunoreaction cell and trapped in it by the neodymium magnet, which was equipped beneath the immunoreaction cell. An APnEOs sample solution containing the HRP-labeled APnEOs at a constant concentration, and a luminol solution containing hydrogen peroxide and p-iodophenol were sequentially introduced into the immunoreaction cell, according to an SIA programmed sequence. Chemiluminescence emission was monitored by means of a photon counting unit located at the upper side of the immunoreaction cell by collecting the emitted light with a lens. A typical sigmoidal calibration curve was obtained, when the logarithm of the concentration of APnEOs was plotted against the chemiluminescence intensity as the number of photons in 100 ms using standard APnEOs sample solutions at various concentrations (0-1000 ppb) under optimum conditions. The lower detection limit defined as IC(80) is ca 10 ppb. The time required for analysis is less than 15 min per a sample. The present method was successfully applied to the determination of APnEOs in river water.. |
| 26. | Koji Nakano, Tadateru Yoshitake, Yasunori Yamashita, Edmond F. Bowden, Cytochrome c Self-Assembly on Alkanethiol Monolayer Electrodes as Characterized by AFM, IR, QCM, and Direct Electrochemistry, Langmuir, https://doi.org/10.1021/la063697w, 23, 11, 6270-6275, 2007.05, With the advantage of carbodiimide coupling chemistry, horse heart cytochrome c (cyt c) has been covalently immobilized onto self-assembled monolayers (SAMs) from 11-mercaptoundecanoic acid (MUDA) developed on single-crystal or polycrystalline gold substrate surfaces. The cyt c immobilized substrates thus prepared have been characterized by atomic force microscopy (AFM); we have succeeded in obtaining surface topographical images down to single-protein resolution. AFM imaging has also shown densely packed, uniform protein monolayer formation that is highly suggestive of self-assembly of cyt c molecules on MUDA SAMs. Covalent attachment of cyt c has been further evidenced by reflection−absorption FT-IR as well as microgravimetric analysis using a quartz crystal microbalance (QCM). In the latter, the specific MUDA and cyt c surface concentrations were determined to be 0.86 ± 0.11 nmol cm-2 (n = 5) and 28 ± 12 pmol cm-2 (n = 5), both of which agree fairly well with their theoretical counterparts. The obtained QCM chips having the cyt c/MUDA/Au interfacial structure were found to be capable of the direct electrochemistry of the surface-attached cyt c molecules. Cyclic voltammetric measurements on the chips gave particular redox waves showing characteristics of surface process. The electroactive protein surface concentration was determined to be 7.2 ± 4.8 pmol cm-2 (n = 6); it was almost consistent with values found in literature, while it was limited to 26% in magnitude for the QCM data. This was deemed to have arisen from the orientation variation of the surface-confined cyt c molecules and is discussed briefly.. |
| 27. | Yan LI, Jujie REN, Hizuru NAKAJIMA, Nobuaki SOH, Koji NAKANO, Toshihiko IMATO, Surface Plasmon Resonance Immunosensor for IgE Analysis Using Two Types of Anti-IgE Antibodies with Different Active Recognition Sites, Analytical Sciences, https://doi.org/10.2116/analsci.23.31, 23, 1, 31-38, Vol. 23, No. 1, pp. 31 ? 38., 2007.01, A simple and novel method for the determination of an IgE antibody based on a surface plasmon resonance immunosensor for the diagnosis of an allergy is described. The method involves the use of an anti-IgE(D) antibody and an anti-IgE(H) antibody, which reacts with the Ce2 domain and the Ce3 domain of the IgE antibody. The anti-IgE(D) antibody was immobilized on the gold surface of a sensor chip by physical adsorption. An IgE antibody sample was incubated by adding it to an anti-IgE(H) antibody solution to form an anti-IgE(H) immunocomplex through a reaction of the Ce3 domain of the IgE antibody. The incubated solution was introduced onto the sensor chip and the immunocomplex of the IgE-anti-IgE(H) then reacted with the anti-IgE(D) antibody immobilized on the sensor chip through the Ce2 domain of the IgE antibody part of the IgE-anti-IgE(H) immunocomplex. The detection limit of the present method for the determination of the IgE antibody was about 10 ppb. The affinity constants for the anti-IgE(H) antibody immunocomplex with the IgE antibody in solution and that of the anti-IgE(H) antibody immunocomplex with the IgE antibody immobilized on the sensor chip by a biotin-streptavidin interaction were estimated to be 4.1 × 107 M-1 and 5.8 × 106 M-1, respectively. The affinity constant for the immunocomplex of the anti-IgE(H) antibody with the IgE antibody with the anti-IgE(D) immobilized on the sensor chip was estimated to be 4.9 × 107 M-1, 20-times larger than the affinity constant for the IgE antibody immunocomplex with the anti-IgE(D) antibody immobilized on the sensor chip, based on a direct immunoassay method of the IgE antibody under the same experimental conditions.. |
| 28. | Koji Nakano, Electrical Transport Through DNA-Ferrocene Conjugate as Characterized by Conductive AFM Measurement, Proceedings of The 2nd International Symposium on Functional Innovation of Molecular Informatics, pp. 27, 2006.11. |
| 29. | K. Nakano, G. Hirayama, M. Toguchi, K. Nakamura, K. Iwamoto, N. Soh, T. Imato, Poly(hydroquinone)-coated electrode for immobilizing of 5’-amine functioned capture probe DNA and electrochemical response to DNA hybridization, Science and Technology of Advanced Materials, https://doi.org/10.1016/j.stam.2006.06.007, 7, 7, 718-725, Vol. 7, No. 7, pp. 718 - 725., 2006.10, Enzymatically polymerized hydroquinone, PHQ, was applied for polymer-coated electrode whose surface was further modified with 5′-amine dodecamer DNAs (capture probe DNA, CP-DNA) by taking advantage of Michael reaction. The film-forming property of PHQ on graphite substrate surfaces was confirmed to be satisfactory by AFM imaging. Cyclic voltammetric (CV) measurements showed that the PHQ-modified graphite electrode gave well-defined redox waves showing characteristics for surface process. The pH dependence of the formal potential, E1/2, suggested that the electrode reaction occurred by two-proton and two-electron mechanism (−59 mV per a pH decade). CVs also gave the specific amount of PHQ adsorption of 0.52 or 0.83 nmol cm−2 (monomer unit) for the different electrode preparations. This was indicative of two- or three-monolayer adsorption of PHQ. For applications of gene detection, the immobilization reaction including the CP-DNA hybridization was studied by microgravimetric analysis using a quartz-crystal microbalance (QCM). Summaries for the two different runs were 1.01 and 0.69 nmol cm−2 (monomer unit) for PHQ adsorption on gold surfaces, 0.19 and 0.15 nmol cm−2 for CP-DNA attachment on the PHQ/Au, and 0.14 and 0.11 nmol cm−2 for hybridization with the complementary DNA on the CP-DNA/PHQ/Au, respectively. Importantly, monitoring of the series of the experiments were possible by measuring the voltammetric properties of the electrode; the distinct redox waves due to PHQ-electrode reaction are suppressed upon immobilizing of the CP-DNA, and its hybridization further suppressed the redox activity. Neither treating of the PHQ-electrode with native DNAs nor treating of CP-DNA/PHQ-electrode with non-target DNA gave no noticeable responses. A possible mechanism for the electrode response was discussed briefly based on electrochemical QCM measurements. We think these observations are important as the basis of DNA hybridization sensor that enables totally, label-free electrochemical detection of the target DNA.. |
| 30. | K. Nakano, H. Matsunaga, K. Sai, N. Soh, T. Imato, Photoactive, covalent attachment of DNA on gold with double-strand specificity using self-assembled monolayers containing psoralen, Anal. Chim. Acta, https://doi.org/10.1016/j.aca.2006.04.085, 578, 1, 93-99, Vol. 578, No. 1, pp.93 - 99, 2006.09, Taking advantages of psoralen photochemistry, we have developed a new method of immobilizing DNA on gold substrate surfaces. A psoralen derivative having an alkylamine function was synthesized, and was self-assembled on gold substrate surfaces in a combined use of a thiol-derivatized molecule, 3,3′-dithiobis(succinimidyl propionate) forming amide bonds on the surface. We found that by irradiating with long wavelength ultraviolet light (320–400 nm), DNA molecules added in the solution phase were covalently immobilized on the monolayer surface through the photoadduct formation of the psoralen molecules with the DNA nucleobases. The present method has its advantage that is applicable to native DNAs, no chemically modifying DNAs, in spite of its covalent immobilization principle. We have examined 12 mer synthetic oligonucleotide immobilizations and have found that the surface concentration thus attained was to be 20 pmol cm−2, which is consistent with saturated surface coverage. Interestingly, the immobilization occurred double-stranded-DNA-preferentially; no immobilization for single-stranded DNAs. Characterization of the immobilization chemistry has been achieved using atomic force microscopic imaging, infrared absorption, X-ray photoelectron spectroscopy, electrochemistry, and quartz-crystal microbalance and their results were described.. |
| 31. | Y. LI, M. KOBAYASHI, K. FURUI, N. SOH, K. NAKANO, T. IMATO, Surface plasmon resonance immunosensor for histamine based on an indirect competitive immunoreaction, Analytica Chimica Acta, https://doi.org/10.1016/j.aca.2006.01.078, 576, 1, 77-83, Vol. 576, No. 1, pp. 77 - 83, 2006.08, he use of a surface plasmon resonance immunosensor for the analysis of histamine (β-imidazole ethylamine) is described. The method is based on an indirect competitive reaction of an anti-histamine antibody in a sample solution with histamine immobilized on a sensor chip and with histamine in the sample solution. A sensor chip immobilized with histamine was prepared using a self-assembly monolayer of 11-mercaptoundecanoic acid (11-MUA) as an anchor membrane, followed by an amino-coupling reaction with histamine after activation of the 11-MUA layer on the sensor chip by treatment with 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and N-hydroxysuccinimide. The sensor chip can be reused, after regeneration with a 10 mM HCl solution, which dissociates the anti-histamine antibody complex from histamine on the sensor chip. The affinity constants for the immunocomplex of the anti-histamine antibody with histamine in the solution and for that of the anti-histamine antibody with histamine immobilized on the sensor chip were calculated to be 1.5 × 107 and 7.2 × 105 M−1, respectively, by assuming a Langmuir-type adsorption of the anti-histamine antibody to histamine immobilized on the sensor chip. The detection limit of the method was determined to be 3 ppb.. |
| 32. | Nobuaki Soh, Daisuke Seto, Koji Nakano, Toshihiko Imato, Novel fuorescent probe for detecting hydroperoxides with strong emission in the visible range, Molecular Biosystems, https://doi.org/10.1016/j.bmcl.2006.02.078, 16, 11, 81-84, Vol. 2, No. 2, pp. 81 - 84, 2006.06, A novel fluorescent probe, a swallow-tailed perylene derivative for detecting hydroperoxides (Spy-HP), containing perylene 3,4,9,10-tetracarboxyl bisimide as the main skeleton in the structure, was developed. Spy-HP reacted rapidly with hydroperoxides such as m-chloroperbenzoic acid (MCPBA) and cumene hydroperoxide to form its oxidized derivative, Spy-HPOx, and emitted an extremely strong fluorescence (Φ ∼ 1) in the visible range (λex = 524 nm and λem = 535 nm), as the result of canceling the photoinduced electron transfer (PET) effect. The reaction between Spy-HP and hydroperoxides proceeded quantitatively in strict stoichiometry, without being affected by autoxidation or photobleaching. Because of these prominent properties, Spy-HP is expected to be a novel and useful fluorescent probe to ‘spy’ on hydroperoxides in biosamples.. |
| 33. | K. Hirakawa, M. Katayama, N. Soh, K. Nakano, H. Ohura, S. Yamasaki, T. Imato, Electrochemical Sandwich Immunoassay for Vitellogenin by Sequential Injection Analysis Using Antibody Immobilized Magnetic Microbeads, Electroanalysis, https://doi.org/10.1002/elan.200603529, 18, 13-14, 1297-1305, Vol. 18, No. 13-14, pp. 1297 - 1305, 2006.06, A rapid and sensitive sandwich immunoassay based on a sequential injection analysis (SIA) for the determination of carp vitellogenin (Vg) is described. The SIA system was constructed from a syringe pump, a multiposition valve, a flow-through type immunoreaction cell equipped with a magnet and an amperometric detector. Magnetic microbeads immobilized with an anti-Vg monoclonal antibody (primary antibody) were used as a solid support. The primary antibody was immobilized on magnetic microbeads by coupling the primary antibody with a polylactic acid-layer, which was coated on the surface of the magnetic beads, after activated with N-hydroxysuccinimide. The introduction, trapping and flushing out of the magnetic microbeads in the immunoreaction cell were controlled by the magnet and the flow of the carrier solution. After the primary antibody-immobilized magnetic beads were introduced and trapped in the immunoreaction cell, a Vg sample solution, an alkaline phosphatase (AP)-labeled anti-Vg polyclonal antibody (secondary antibody) solution and a p-aminophenyl phosphate (PAPP) solution were sequentially introduced into the immunoreaction cell based on an SIA programmed sequence. Vg was determined by the electrochemical detection of p-aminophenol (PAP), an enzymatic product of PAPP via the action by AP labeled on the secondary antibody. A solution containing PAP, which was generated in the immunoreaction cell and transiently held in a holding coil, was transported to the amperometric detector and the oxidation current of PAP on a working electrode applied at +0.20 V was measured. A sigmoidal calibration curve was obtained in the concentration range from 1 ppb to 500 ppb in a plot of oxidation current against the logarithm of the Vg concentration. The lower detection limit of the immunoassay was about 2–3 ppb. The time required for an analysis was ca. 15 min/sample.. |
| 34. | N. SOH, D. SETO, K. NAKANO, T. IMATO, Methodology of reversible protein labeling for ratiometric fluorescent measurement, Molecular BioSystems, DOI https://doi.org/10.1039/B515777C, 2, 2, 128-131, Vol. 2, No. 2, pp. 128 - 131, 2006.02, The first fluorescent labeling technology, which can induce not only an increase in the fluorescence intensity but also a shift in the fluorescence spectrum, has been developed for “ratiometric” measurements for a protein by utilizing a newly designed “field-sensitive” fluorescent probe and its corresponding unique amino acid tag.. |
| 35. | K. HIRAKAWA, M. KATAYAMA, N. SOH, K. NAKANO, T. IMATO, Electrochemical Immunoassay for Vitellogenin Based on Sequential Injection Using Antigen-immobilized Magnetic Microbeads, Analytical Sciences, https://doi.org/10.2116/analsci.22.81, 22, 1, 81-86, Vol 22, No. 2, pp.81 - 86, 2006.01, A rapid and sensitive immunoassay for the determination of vitellogenin (Vg) is described. The method involves a sequential injection analysis (SIA) system equipped with an amperometric detector and a neodymium magnet. Magnetic beads, onto which an antigen (Vg) was immobilized, were used as a solid support in an immunoassay. The introduction, trapping and release of magnetic beads in an immunoreaction cell were controlled by means of the neodymium magnet and by adjusting the flow of the carrier solution. The immunoassay was based on an indirect competitive immunoreaction of an alkaline phosphatase (ALP) labeled anti-Vg monoclonal antibody between the fraction of Vg immobilized on the magnetic beads and Vg in the sample solution. The immobilization of Vg on the beads involved coupling an amino group moiety of Vg with the magnetic beads after activation of a carboxylate moiety on the surface of magnetic beads that had been coated with a polylactate film. The Vg-immobilized magnetic beads were introduced and trapped in the immunoreaction cell equipped with the neodymium magnet; a Vg sample solution containing an ALP labeled anti-Vg antibody at a constant concentration and a p-aminophenyl phosphate (PAPP) solution were sequentially introduced into the immunoreaction cell. The product of the enzyme reaction of PAPP with ALP on the antibody, p-aminophenol, was transported to an amperometric detector, the applied voltage of which was set at +0.2 V vs. an Ag/AgCl reference electrode. A sigmoid calibration curve was obtained when the logarithm of the concentration of Vg was plotted against the peak current of the amperometric detector using various concentrations of standard Vg sample solutions (0 - 500 ppb). The time required for the analysis is less than 15 min.. |
| 36. | Koji NAKANO, Hideshi MATSUNAGA, Keisuke SAI, Covalent attachment of DNA with double-strand specificity using self-assembled monolayers from psoralen, Proceedings of The 2005 International Chemical Congress of Pacific Basin Societies, pp. 366, 2005.12. |
| 37. | Hideshi MATSUNAGA, Koji NAKANO, Nobuaki SOH, Toshihiko IMATO, Preparation, Characterization and Atomic Force Microscopic Imaging of New DNA Ligand for Single Molecular Hybridization Assay, Proceedings of The 2005 International Chemical Congress of Pacific Basin Societies, pp. 968, 2005.12. |
| 38. | Koji NAKANO, Go HIRAYAMA, Mikiko TOGUCHI, Kaori NAKAMURA, Kaori IWAMOTO, Nobuaki SOH, Toshihiko IMATO, DNA Conjugate Electrode Having DNA/Oligo(1,4-hydroquinone)/PG Interfacial Structure for Biosensor Applications: Characterization by Electrochemical Measurements, QCM Analysis, and AFM Imaging, Program & Abstracts for the 8th Asian Conference on Analytical Sciences, pp. 138, 2005.10. |
| 39. | R-Q. Zhang, K. Hirakawa, D. Seto, N. Soh, K. Nakano, T. Masadome, K. Nagata, K. Sakamoto, T. Imato, Sequential injection chemiluminescence immunoassay for anionic surfactants using magnetic microbeads immobilized with an antibody., Talanta, 10.1016/j.talanta.2005.07.012, 68, 2, 231-238, VOL. 68, No. 2, pp. 231 - 238, 2005.10, A rapid and sensitive immunoassay for the determination of linear alkylbenzene sulfonates (LAS) is described. The method involves a sequential injection analysis (SIA) system equipped with a chemiluminescence detector and a neodymium magnet. Magnetic beads, to which an anti-LAS monoclonal antibody was immobilized, were used as a solid support in an immunoassay. The introduction, trapping and release of the magnetic beads in the flow cell were controlled by means of a neodymium magnet and adjusting the flow of the carrier solution. The immunoassay was based on an indirect competitive immunoreaction of an anti-LAS monoclonal antibody on the magnetic beads and the LAS sample and horseradish peroxidase (HRP)-labeled LAS, and was based on the subsequent chemiluminscence reaction of HRP with hydrogen peroxide and p-iodophenol, in a luminol solution. The anti-LAS antibody was immobilized on the beads by coupling the antibody with the magnetic beads after activation of a carboxylate moiety on the surface of magnetic beads that had been coated with a polylactic acid film. The antibody immobilized magnetic beads were introduced, and trapped in the flow cell equipped with the neodymium magnet, an LAS solution containing HRP-labeled LAS at constant concentration and the luminol solution were sequentially introduced into the flow cell based on an SIA programmed sequence. Chemiluminescence emission was monitored by means of a photon counting unit located at the upper side of the flow cell by collecting the emitted light with a lens. A typical sigmoid calibration curve was obtained, when the logarithm of the concentration of LAS was plotted against the chemiluminescence intensity using various concentrations of standard LAS samples (0–500 ppb) under optimum conditions. The time required for analysis is less than 15 min..  |
| 40. | T. TSURUTA, Y. KATSUMI, S. SHIRAKAWA, S. TAGUCHI, K. NAKANO, Synthesis of New Psoralen Derivatives for Chemical Modification and Functionalization of DNA Duplexes as Characterized by Gel Electrophoresis, IR, QCM, Electrochemistry, and AFM, Abstract Book of International Symposium on Nano-organization and Function, 2004.11. |
| 41. | H. MATSUNGAE, K. SAI, K. NAKANO, Covalent Attachment of Double-Stranded DNA using Self-Assembled Monolayers Containing Psoralen as Characterized by XPS, IR, QCM, Electrochemistry, and AFM, Abstract Book of International Symposium on Nano-organization and Function, pp. 48, 2004.11. |
| 42. | K. NAKANO, K. DOI, K. TAMURA, Y. KATSUMI, Monolayer Formation of Glucose Oxidase Covalently Attached on ω-Aminoalkanethiol/Au via Activation of Carbohydrate Residue of GOx as Characterized by AFM, IR, QCM, and Biosensor Applications, Abstract of The 7th Asian Conference on Analytical Sciences, pp. 308, 2004.10. |
| 43. | T. Anada, H. Matsunaga, R. Karinaga, K. Koumoto, M. Mizu, K. Nakano, S. Shinkai, and K. Sakuraia, Proposal of new modification technique for linear double-stranded DNAs using the polysaccharide schizopyllan, Bioorganic & Medicinal Chemistry Letters, 10.1016/j.bmcl.2004.08.043, 14, 22, 5655-5659, Vol.14, pp.5655 - 5659, 2004.09. |
| 44. | K. Nakano, S. Shirakawa, S. Taguchi, A psoralen Derivative Containing Ferrocene for Redox-labeling of DNA and Electrochemical Gene Sensor Applications, Chemcial Sensors, Vol. 20, Supplement B, pp. 778 - 779., 2004.07. |
| 45. | Koji Nakano, DNA-Ferrocene Conjugate for Nanoelectronic Material Applications. Atomic Force Microscope-Based, Direct Measurement of Electrical Transport through DNA Molecules, Second 21st Century COE “Towards Creating New Industries Based on Inter-Nanoscience” and 7th SANKEN International Symposium on “Hybridization of Chemistry, Biology, and Material Science”, pp. 38 - 39, 2004.01. |
| 46. | K. Nakano, K. Doi, K. Tamura, Y. Katsumi, Self-Assembling Monolayer Formation of Glucose Oxidase and Cytochrome c Covalently Attached on Alkanethiol Monolayers on Gold, Biomolecular Chemistry. Proceedings of the ISBC 2003, pp. 284 - 285, 2003.12. |
| 47. | K. NAKANO, K. DOI, K. TAMURA, Y. KATSUMI, Self-assembling Monolayer Formation of Glucose Oxidase Covalently Attached on 11-Aminoundecanethiol Monolayers on Gold, Chemical Communications, 10.1039/b303298a, 13, 1544-1545, Vol. 2003, No. 13, pp. 1544 - 1545, 2003.06, Glucose oxidase (GOx) has been attached covalently to form uniform enzyme monolayers on self-assembled monolayers (SAMs) from 11-aminoundecanethiol (AUDT) by taking advantage of chemical oxidation of GOx carbohydrate residues followed by coupling the resulting ‘aldehydic’ enzyme with the terminal amino group in the SAM as characterized by AFM imaging, IR, QCM, and electrochemical measurements. . |
主要総説, 論評, 解説, 書評, 報告書等
主要学会発表等
作品・ソフトウェア・データベース等
| 1. | 中野幸二, 「ぶんせき」誌2005年度表紙写真, 2005.01 日本分析化学会のメインの機関誌である「ぶんせき」について、2005年度の表紙写真として実験データが採択された。. |
学会活動
学協会役員等への就任
2016.04～2018.03, 日本分析化学会, 代議員.
2007.10～2008.09, 日本分析化学会, オンライン登録委員会.
2006.04～2007.03, 日本分析化学会, 九州支部副支部長.
2004.02～2007.01, 電気化学会, 九州支部幹事.
2003.04～2004.03, 日本分析化学会, 広報委員会年会・討論会オンライン登録委員.
2002.04～2003.03, 日本分析化学会, 九州支部副支部長.
1995.04, 日本分析化学会, 九州支部幹事.
1997.04～1999.03, 電気化学会, 九州支部庶務幹事.
学会大会・会議・シンポジウム等における役割
2019.05.18～2019.05.19, 第79回分析化学討論会, 座長（Chairmanship）.
2018.09.12～2018.09.14, 日本分析化学会第67年会, 座長（Chairmanship）.
2017.05.27～2017.05.28, 日本分析化学会第77回分析化学討論会, 座長（Chairmanship）.
2017.05.05～2017.05.08, International Congress on Analytical Sciences (ICAS2017), 座長（Chairmanship）.
2015.09.09～2015.09.11, 日本分析化学会第64年会, 座長（Chairmanship）.
2014.09.17～2014.09.19, 日本分析化学会第63年会, 座長（Chairmanship）.
2013.09.10～2012.09.12, 日本分析化学会第62年会, 座長（Chairmanship）.
2013.08.22～2013.08.24, The Twelfth Asian Conference on Analytical Sciences (ASIANALYSIS XII), 座長（Chairmanship）.
2011.09.14～2011.09.16, 日本分析化学会第60年会, 座長（Chairmanship）.
2008.09.10～2008.09.12, 日本分析化学会第57年会, 座長（Chairmanship）.
2008.05.15～2008.05.16, 第69回分析化学討論会, 座長（Chairmanship）.
2007.09, 日本分析化学会第56年会, 座長（Chairmanship）.
2007.05, 第68回分析化学討論会, 座長（Chairmanship）.
2007.03, 日本化学会第87春季年会, 座長（Chairmanship）.
2006.09, 日本分析化学会第55年会, 座長（Chairmanship）.
2005.09, 日本分析化学会第54年会, 座長（Chairmanship）.
2005.05, 第66回分析化学討論会, 座長（Chairmanship）.
2004.11, International Symposium on Nano-organization and Function, 座長（Chairmanship）.
2004.05, 第65回分析化学討論会, 座長（Chairmanship）.
2004.03, 日本化学会第84春季年会, 座長（Chairmanship）.
2003.09, 日本分析化学会第52年会, 座長（Chairmanship）.
2003.10, 第1回生体機能関連化学・バイオテクノロジー部会合同シンポジウム, 座長（Chairmanship）.
2002.03, 日本化学会第81春季年会, 座長（Chairmanship）.
2015.09.09～2015.09.11, 日本分析化学会第57年会, 実行委員.
2015.09.02～2015.09.04, 平成27年度工学研究教育講演会, 実行委員.
2014.11.30～2014.12.05, 19th International Conference on Flow Injection Analysis and Related Techniques, Organizing Committee.
2013.08.22～2013.08.23, The Twelfth Asian Conference on Analytical Sciences (ASIANALYSIS XII), Organizing Committee.
2012.05.19～2012.05.20, 第72回分析化学討論会, 実行委員.
2007.10, 日本分析化学会第57年会, 実行委員.
2005.04, 電気化学会第72回大会, 実行委員.
2004.11, International Symposium on Nano-organization and Function, 実行委員.
2003.05, 第65回分析化学討論会, 実行委員.
2001.11, 日本分析化学会第50年会, 実行委員.
1998.07, The 49th Annual Meeting of The International Society of Electrochemistry, Organizing Committe.
1998.10, 工業物理化学講習会, 庶務幹事.
1997.10, 工業物理化学講習会, 庶務幹事.
1997.05, 第58回分析化学討論会, 実行委員.
1995.07, 分析化学講習会, 庶務幹事.
学術論文等の審査
| 年度 | 外国語雑誌査読論文数 | 日本語雑誌査読論文数 | 国際会議録査読論文数 | 国内会議録査読論文数 | 合計 |
|---|---|---|---|---|---|
| 2023年度 | 2 | 0 | 0 | 0 | 2 |
| 2022年度 | 5 | 0 | 0 | 0 | 5 |
| 2021年度 | 5 | 1 | 0 | 0 | 6 |
| 2020年度 | 5 | 0 | 0 | 0 | 5 |
| 2019年度 | 2 | 0 | 0 | 0 | 2 |
| 2018年度 | 10 | 0 | 0 | 0 | 10 |
| 2017年度 | 10 | 0 | 0 | 0 | 10 |
| 2015年度 | 10 | 0 | 0 | 10 | |
| 2014年度 | 20 | 0 | 0 | 20 | |
| 2013年度 | 20 | 0 | 0 | 20 | |
| 2012年度 | 20 | 0 | 0 | 0 | 20 |
| 2011年度 | 10 | 0 | 0 | 0 | 10 |
| 2010年度 | 10 | 0 | 0 | 0 | 10 |
| 2009年度 | 15 | 0 | 0 | 0 | 15 |
| 2008年度 | 15 | 0 | 0 | 0 | 15 |
| 2007年度 | 15 | 5 | 0 | 0 | 20 |
| 2006年度 | 6 | 1 | 5 | 0 | 6 |
| 2005年度 | 10 | 5 | 0 | 0 | 15 |
| 2004年度 | 10 | 5 | 20 | 0 | 35 |
| 2003年度 | 8 |
その他の研究活動
海外渡航状況, 海外での教育研究歴
Sheraton Haikou Hotel, Haikou, China, 2017.05.
The Empress Hotel, Chiang Mai, Thailand, 2016.12.
Ramada Plaza Hotel, Jeju, Korea, 2015.12.
Hawaii University at Manoa, UnitedStatesofAmerica, 2012.03～2012.03.
Hawaii Convention Center, UnitedStatesofAmerica, 2010.12.
Kumoh National Institute of Technology, Korea, 2010.09.
University of Edinburgh, UnitedKingdom, 2010.09.
Ramada Plaza Jeju, Korea, 2007.11.
North Carolina State University, UnitedStatesofAmerica, 1995.10～1996.09.
受賞
フローインジェクション分析論文賞, 日本分析化学会・フローインジェクション分析研究懇談会, 2013.11.
フローインジェクション分析論文賞, 日本分析化学会・フローインジェクション分析研究懇談会, 2006.06.
奨励賞, 日本分析化学会, 1996.09.
若手研究論文「化学のフロンティアIX」採択, 日本化学会, 1993.02.
研究資金
科学研究費補助金の採択状況(文部科学省、日本学術振興会)
2022年度～2026年度, 基盤研究(S), 分担, 匂いの時空間揺らぎ情報に基づく 人探索.
2020年度～2022年度, 基盤研究(B), 代表, 塩基配列選択的なDNA二重らせん結合性フォトンアップコンバージョン蛍光プローブ.
2018年度～2020年度, 基盤研究(A), 分担, 匂いイメージセンサによる匂い痕跡画像の要素臭プロファイル分解.
2015年度～2017年度, 基盤研究(A), 分担, 匂いの質と空間の可視化センシング.
2013年度～2014年度, 挑戦的萌芽研究, 代表, 痛み情報の化学計測とin vivo蛍光イメージング.
2009年度～2011年度, 新学術領域研究, 代表, ＤＮＡチップエレクトロニクスの創製とナノバイオセンシング.
2009年度～2011年度, 基盤研究(C), 代表, ＤＮＡ自己組織化膜のｉｎ－ｓｉｔｕ電気回路化を利用するチップ型遺伝子センサー.
2003年度～2004年度, 基盤研究(C), 代表, ＤＮＡコンジュゲートを感応素子に用いるバイオエレクトロケミカルアレイ型センサ.
2000年度～2001年度, 基盤研究(C), 代表, 遺伝子ターゲッティングツールとしてのプラスミドDNA-フェロセン複合体の研究.
競争的資金(受託研究を含む)の採択状況
2012年度～2013年度, 科学技術振興機構 復興促進プログラム（A-STEP）探索タイプ, 代表, ＤＮＡ修復の電気化学検出を利用する遺伝子の放射線変異センシング.
2011年度～2013年度, 平成23年度戦略的情報通信研究開発推進制度, 分担, 匂いイメージセンサによる情報創出に関する研究開発.
2009年度～2009年度, 科学技術振興機構 シーズ発掘試験, 代表, 自立性自家応答性遺伝子センサー.
2008年度～2009年度, 九州大学先端融合COE・先端融合医療レドックスナビ研究拠点研究テーマ , 代表, 光誘起電子移動による蛍光スイッチング機能を組み込んだ分子認識性量子ドットセンサーとバイオイメージング応用.
2005年度, 4) 科学技術振興機構 研究成果活用プラザ福岡 「実用化のための可能性試験, 代表, １分子検出のための遺伝子プローブ−DNAリガンド−の研究.
2000年度～2003年度, 科学技術振興機構戦略的創造研究推進事業「さきがけ２１」, 代表, DNA二重らせんを電子機能・構造単位とする単一分子素子.
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九大関連コンテンツ
QIR 九州大学学術情報リポジトリ システム情報科学研究院
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