旭硝子財団助成研究成果報告2018

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Design and synthesis of novel lipophilic fluorescent substrate and development of capillary-type single-step immuno-diagnostics based on the clarification of the enzyme reaction mechanism at the oil-water interface (Project 2016)38久本　秀明33Hideaki HISAMOTO平岡　秀一34Shuichi HIRAOKA新規疎水性蛍光基質分子群の設計・合成と油水界面酵素反応機構解明に基づくキャピラリー型1ステップ免疫診断の展開(2016採択)自己集合性超分子マクロサイクルの形成メカニズムの解明(2016採択)Investigation of Formation Mechanism of Self-assembled Supramolecular Macrocycles(Project 2016)旭硝子財団　助成研究成果報告（2018）本研究では油水界面で酵素反応を検出できる疎水性色素の開発，およびその反応性評価，１ステップバイオアッセイデバイスへの応用を目的とした．その結果，ペルオキシダーゼを検出可能な疎水性酵素基質分子の開発に成功した．また，酵素反応を検出できる疎水性色素を含む可塑化PVC膜の作製過程で超高感度かつ高速な応答が期待できるイオン液体型色素を見出した．最終的に１ステップバイオアッセイデバイスをデザインし，これまで困難だった可塑化PVC膜を均一に固定化する方法開発にも成功した．本研究の成果は，今後イムノアッセイ，バイオセンシング等が可能な，多様な１ステップバイオアッセイデバイス開発への展開が期待できる．In this study, we developed a hydrophobic dye capable of detecting an enzymatic reaction at the or-ganic-aqueous interface, aimed at its reactivity evaluation and its application to a single step bioassay device. As a result, we successfully developed a hydrophobic enzyme substrate molecule capable of detecting horse raddish peroxidase. We have also found an ionic liquid-based dye which can be expec-ted to have ultrahigh sensitivity and high speed response in the process of preparing a plasticized PVC membrane containing a hydrophobic dye capable of detecting an enzymatic reaction. Finally, we also succeeded in developing a method of uniformly immobilizing plasticized PVC membrane on a single step bioassay device. The results of this research can be applied to develop various single-step bioassay devices capable of immunoassay, biosensing, etc. in the future.自己集合は構成要素が自発的に集まり，秩序構造を形成する現象である．自己集合は自然界で多々観測されるが，その形成機構は殆ど明らかにされていなかった．これは一過的に生成する中間種の観測が極めて難しいことが主な原因である．筆者らはこの問題を解決するために，中間体に比べずっと観測が容易な原料及び生成物を全て定量することにより，全中間体の平均組成を求め，この時間発展から自己集合の形成機構を明らかにする新手法(QASAP: quantitative analysis of self-assembly process)を開発した．本研究では，QASAPを自己集合性環状金属錯体に適用し，形成機構を明らかにすることに成功した．Self-assembly is a phenomenon that components spontaneously assemble into a well-ordered struc-ture and many examples are seen in Nature. However, self-assembly processes have not been fully un-derstood yet. This is mainly due to difficulty in the observation of intermediates transiently produced during the self-assembly. In order to solve this problem, we have recently developed a method for the investigation of molecular self-assembly (QASAP: quantitative analysis of self-assembly process), where the quantification of all the substrates and the products, which is much easier than that of the interme-diates, enables us to obtain the average composition of all the intermediates, whose time development leads to the self-assembly process. In this research, we applied QASAP to macrocyclic coordination self-assemblies and succeeded in revealing their self-assembly processes.