Signaling protein-imprinted polymer array having a fluorescent dye located within the imprinted cavity as a reporter molecule for the binding events

信号蛋白印迹聚合物阵列具有位于印迹空腔内的荧光染料作为结合事件的报告分子

• 批准号: 24651261
• 负责人: TAKEUCHI Toshifumi
• 金额: $ 2.58万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24651261/
• 关键词: 分子インプリンティング; 分子認識; タンパク質; 生体分子計測

A linker molecule comprised of a disulfide linkage to immobilize proteins for the preparation of signaling protein-imprinted polymers on substrates was designed and synthesized. The proteins coupled with the linker molecule maintained their secondary structure, and the molecular imprinting process properly proceeded. After the polymerization, the immobilized proteins were easily removed by the reduction of the disulfide moiety to leave the imprinted cavities capable of introducing thiol-reactive fluorescent dyes for signal transduction of binding events. Thickness of the protein imprinted polymer thin layer on the substrate was examined and, it is revealed that 10 to 20 nm thickness was suitable for the molecular recognition. Post-imprinting modification technique was developed to reduce the number of low affinity binding cavities, resulting in the improvement of S/N ratio.

设计并合成了一种由二硫键连接到信号蛋白的连接分子，用于制备底物上的信号蛋白印迹聚合物。与连接分子偶联的蛋白质保持其二级结构，并且分子印迹过程正确地进行。聚合后，固定化的蛋白质很容易被去除的二硫化物部分的还原，留下的印迹腔能够引入巯基反应性荧光染料的信号转导的结合事件。检测了基质上蛋白质印迹聚合物薄层的厚度，发现10至20 nm的厚度适合于分子识别。采用印迹后修饰技术，减少了低亲和力结合空穴的数量，提高了信噪比。

项目成果

Protein imprinting by living radical polymerization

通过活性自由基聚合进行蛋白质印迹

• 发表时间: 2012
• 作者: Kamon Y.;Ooya;T.;Takeuchi;T.
• 通讯作者: T.

Post- imprinting treatment of molecularly imprinted polymers for proteins to enhance specific binding signals

蛋白质分子印迹聚合物的印后处理以增强特异性结合信号

• 发表时间: 2012
• 作者: Sunayama,H.;Ooya;T.;Takeuchi;T.
• 通讯作者: T.

表面開始AGET ATRPによるタンパク質インプリンティング

表面引发的 AGET ATRP 蛋白质印迹

• 发表时间: 2013
• 作者: 大塚康平;松岡茂;廣瀬未果;佐藤文憲;溝端栄一;杉山成;井上豪;村田道雄;香門悠里,北山雄己哉,大谷亨,竹内俊文
• 通讯作者: 香門悠里,北山雄己哉,大谷亨,竹内俊文

高親和性タンパク質インプリント空間への選択的蛍光レポーター分子の導入

将选择性荧光报告分子引入高亲和力蛋白质印迹空间

• 发表时间: 2013
• 作者: 砂山博文,大谷亨,竹内俊文

Molecularly imprinted polymers prepared using protein-conjugated cleavable monomers followed by site-specific post-imprinting introduction of fluorescent reporter molecules

• DOI: 10.1039/c3cc40484f
• 发表时间: 2013-01-01
• 期刊: CHEMICAL COMMUNICATIONS
• 影响因子: 4.9
• 作者: Suga, Yusuke;Sunayama, Hirobumi;Takeuchi, Toshifumi
• 通讯作者: Takeuchi, Toshifumi