56萩原 正規Masaki HAGIHARA高井まどかMadoka TAKAI核酸型タンパク質捕捉素子の開発とタンパク質検出デバイスへの展開 (2014年採択)Development of a practical protein-chip using designed DNA-arrays (Project 2014)生体親和性蛍光ラベル化高分子ナノ粒子を用いた細胞内温度計測プローブ創製 (2014年採択)Development of cell analysis thermoprobe by use of fluorescence label biocompatible nano-particles (Project 2014)23in recent years, metal-dielectric nano-units gathered much attention in response to reports that various optical properties can be enhanced by coupling of dielectrics and metals. Therefore, the applicant challenged to pre-pare metal nanostructures that has uniform shape and periodically arranged by a bottom-up method suitable for mass production.本研究提案は，臨床現場で使用できるレベルの簡便性，迅速性，高い検査精度，再現性，低コスト化等を視野にいれた，優れたタンパク質発現解析ツール（タンパク質検出デバイス）創製を目標とする．今回，既存のプロテインチップ創製法とは全く異なるグアニン塩基に富む「核酸構造体ライブラリー」をタンパク質認識素子として利用できることを示した．This research proposal aims to create a superior protein analysis tool (protein detection device) with a view-point of convenience, quickness, high inspection accuracy, reproducibility, low cost etc. at a level that can be used in a clinical purpose. Here, we showed that "nucleic acid structure library", that is rich in guanine bases, can serve as a noble protein recognition element.細胞内部の化学反応による熱産生のイメージングは細胞の状態や異常を検知する重要な指標であり，簡便で高精度な細胞内温度イメージング手法が求められている．本研究では，生体親和性を示す 2-meth-acryloyloxyethyl phosphorylcholine(MPC)，疎水部安定のためのn-butylmethacrylate(BMA)，温度応答性蛍光分子のmethacryloxyethyl thiocarbonyl rhodamine B(Rho)および温度不感応な蛍光分子(Cou)から，ブロックコポリマーpoly(MPC)-block-(BMA-co-Rho)-Couを合成した．次にpoly(MPC)-block-(BMA-co-Rho)-Couから約25nmのナノ粒子を作製した．このナノ粒子の二蛍光の強度比から温度変化を計測でき，ナノ粒子は細胞内に容易に取込まれ，細胞内温度計測の可能性が示唆された．Imaging of thermogenesis by chemical reaction inside the cell is an imporatant indicator for detecting the state and abnormality of the cell. For the purpose of the temperature imaging, simple and highly accurate in-tracellular temperature imaging method is required. In this study, we prepared the block copoplymer: poly(MPC)-block-(BMA-co-Rho)-Cou, consisting of biocompatible unit of 2-methacryloyloxyethyl phospho-rylcholine(MPC), hydrophobic unit of n-butylmethacrylate(BMA), temperature responsiveness fluorescent molecules unit of methacryloxyethyl thiocarbonyl rhodamine B(Rho), temperature insensitive fluorescent molecules unit (Cou). Nanoparticles formed by poly(MPC)-block-(BMA-co-Rho)-Cou showed the tempera-ture dependence measuring by the ratio of fluorescent intensity of Rho and Cou,and the diameter of the nano-particle is around 25 nm. Nanoparticles were easily uptaken into the cells, it is suggested that the method of nanoparticle has an ability of the intracellular temperature imaging.
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