An enzyme-immobilization method for integration of biofunctions on a microchip using a water-soluble amphiphilic phospholipid polymer having a reacting group
An enzyme-immobilization method for integration of biofunctions on a microchip using a water-soluble amphiphilic phospholipid polymer having a reacting group
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DOI：
10.1039/b310932a
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发表时间：
2004-01-01
期刊：
影响因子：
6.1
通讯作者：
Toyo'oka, T
中科院分区：
文献类型：
--
作者：
Sakai-Kato, K;Kato, M;Toyo'oka, T
在侧链中具有活性酯基的水溶性磷脂聚合物，聚[2-甲基丙烯酰氧基乙基磷酰胆碱（MPC）-co-甲基丙烯酸正丁酯（BMA）-co-对硝基苯氧基羰基聚乙二醇甲基丙烯酸酯（MEONP）（PMBN），用于将酶固定在塑料微芯片上。将具有BMA单元的MPC聚合物吸附到聚（甲基丙烯酸甲酯）（PMMA）微芯片上，并且MEONP单元中的活性酯基团与蛋白水解酶胰蛋白酶的氨基反应。胰蛋白酶被固定在样品池上，并催化荧光标记的ArgOEt水解为Arg。随后从底物中分离产物及其检测都集成在微芯片上，这意味着从酶活性到产物检测的所有程序都在不到三分钟的时间内完成。
A water-soluble phospholipid polymer having an active ester group in the side chain, poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-nitrophenyloxycarbonyl polyethyleneglycol methacrylate (MEONP) ( PMBN), was used for the immobilization of an enzyme on a plastic microchip. The MPC polymers with BMA units were adsorbed onto the poly( methyl methacrylate) ( PMMA) microchip, and the active ester group in the MEONP unit reacted with the amino groups of the proteolytic enzyme, trypsin. Trypsin was immobilized on the sample reservoir, and catalyzed the hydrolysis of the fluorescently labeled ArgOEt to Arg. The consequent separation of product from the substrate, and their detection, were integrated on the microchip and this meant that all procedures from the enzymatic activity to product detection were completed in less than three minutes.