Development of high-performance blotting device based on microscale electrophoresis

基于微量电泳的高性能印迹装置的研制

• 批准号: 24750068
• 负责人: SUEYOSHI Kenji
• 金额: $ 2.91万
• 依托单位: Osaka Prefecture University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 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/grant/KAKENHI-PROJECT-24750068/
• 关键词: キャピラリー電気泳動; マイクロチップ電気泳動; ブロッティング分析; タンパク質分析; ゲル電気泳動; アフィニティ電気泳動; アルギン酸ヒドロゲル

A novel protein analysis using alginate hydrogels was developed. The alginate hydrogels were easily prepared and immobilized in capillaries and microchannels. It was confirmed that the prepared hydrogel was used as molecular sieving matrix. When the DNAs were introduced into the capillary filled with the alginate hydrogel, some DNAs were separated during the migration in the formed hydrogel, the others were trapped. The sieving effect was controllable by varying concentration of alginate and calcium ions. It was also confirmed that the alginate hydrogels was applicable to affinity capillary electrophoresis. In the experiments, the alginate hydrogel containing avidin specifically captured biotinylated molecules, resulting in the selective separation and preconcentration of the target molecules. Consequently, it is clarified that the alginate hydrogel is versatile material to realize blotting device based on microscale electrophoresis.

开发了一种新的利用海藻酸钠水凝胶进行蛋白质分析的方法。海藻酸钠水凝胶易于制备并固定在毛细管和微通道中。证实了所制备的水凝胶用作分子筛基质。当DNA被引入到填充有海藻酸钠水凝胶的毛细管中时，一些DNA在形成的水凝胶中迁移过程中被分离，另一些则被捕获。通过改变海藻酸钠和钙离子的浓度可以控制筛分效果。同时也证实了海藻酸钠水凝胶适用于亲和毛细管电泳。在实验中，含有抗生物素蛋白的藻酸盐水凝胶特异性地捕获生物素化的分子，导致目标分子的选择性分离和预浓缩。因此，它是澄清，海藻酸钠水凝胶是通用的材料，以实现基于微尺度电泳印迹装置。

项目成果

Effect of a Low-conductivity Zone on Field-amplified Sample Stacking in Microchip Micellar Electrokinetic Chromatography

微芯片胶束电动色谱中低电导率区对场放大样品堆积的影响

• DOI: 10.2116/analsci.29.133
• 发表时间: 2013
• 期刊: Analytical Sciences
• 影响因子: 1.6
• 作者: 末吉健志;北川文彦;大塚浩二
• 通讯作者: 大塚浩二

Development of a capillary partially filled with an affinity ligand-encapsulated hydrogel for electrophoresis. 2

开发部分填充有亲和配体封装水凝胶的毛细管，用于电泳。

• 发表时间: 2012
• 作者: Yudai Fukushima;Kenji Sueyoshi;Fumihiko Kitagawa;Koji Otsuka
• 通讯作者: Koji Otsuka

アルギン酸ヒドロゲルを分離媒体として用いる電気泳動分析

海藻酸盐水凝胶作为分离介质的电泳分析

• 发表时间: 2012
• 作者: 井川真宏;末吉健志;久保拓也;北川文彦;大塚浩二
• 通讯作者: 大塚浩二

Zone electrophoresis of proteins in poly(dimethylsiloxane) (PDMS) microchip coated with physically adsorbed amphiphilic phospholipid polymer

• DOI: 10.1007/s10404-012-1102-8
• 发表时间: 2013-06
• 期刊: Microfluidics and Nanofluidics
• 影响因子: 2.8
• 作者: Kyosuke Nii;K. Sueyoshi;K. Otsuka;M. Takai

Design of Single-Step Immunoassay Principle Based on the Combination of Enzyme-Labeled Antibody Release Coating and Hydrogel Copolymerized with Fluorescent Enzyme Substrate in a Microfluidic Capillary Device

微流控毛细管装置中基于酶标记抗体释放涂层和荧光酶底物共聚水凝胶组合的单步免疫测定原理设计

• 发表时间: 2013
• 期刊: Lab on a Chip
• 影响因子: 6.1
• 作者: Hideki Wakayama;Terence Henares;Kaede Jigawa;Shun-ichi Funano;Kenji Sueyoshi;Tatsuro Endo;Hideaki Hisamoto
• 通讯作者: Hideaki Hisamoto