Rapid measurement of transaminase activities using an amperometric l-glutamate-sensing electrode based on a glutamate oxidase–polyion complex-bilayer membrane
Rapid measurement of transaminase activities using an amperometric l-glutamate-sensing electrode based on a glutamate oxidase–polyion complex-bilayer membrane
复制标题
使用基于谷氨酸氧化酶-聚离子复合双层膜的安培 L-谷氨酸传感电极快速测量转氨酶活性
DOI：
10.1016/s0925-4005(98)00251-2
复制
发表时间：
1998
影响因子：
8.4
通讯作者：
S. Iijima
中科院分区：
文献类型：
--
作者：
F. Mizutani;Yukari Sato;T. Sawaguchi;S. Yabuki;S. Iijima
将谷氨酸氧化酶固定在多离子络合修饰电极上，制备了L谷氨酸电流型传感电极。首先，将3-巯基丙酸浸泡在含有改性剂的乙醇溶液中，在金电极表面生成单层3-巯基丙酸。然后，将聚L赖氨酸和聚(4-苯乙烯磺酸)的水溶液依次放置在电极表面并晾干。最后，通过添加戊二醛溶液使酶交联，在poly-l-lysine/poly(4-styrenesulfonate)-complex层上形成了一层谷氧合酶。基于≈-100分子截留的基于溶质大小的渗透选择性的双层内层多离子复合膜的使用，为L谷氨酸传感电极带来了高性能特性；它具有高灵敏度(检测下限，20 nm)、快速响应(100%响应时间，3 S)、低干扰水平(L-抗坏血酸与相同浓度的L-谷氨酸的响应比，8×10−~2)和高稳定性(可使用一个多月以上)。该双层电极可用于血清中谷草转氨酶和谷丙转氨酶的快速测定：每种转氨酶(0.2-1000U L−1)均可在10 S范围内检测到。
An amperometric l-glutamate-sensing electrode was prepared by immobilizing glutamate oxidase (GlOx) on a polyion complex layer-modified electrode. First, a monolayer of 3-mercaptopropionic acid was made on the surface of a gold electrode by immersing it in an ethanol solution containing the modifier. Next, aqueous solutions of poly-l-lysine and poly(4-styrenesulfonate) were successively placed on the electrode surface and allowed to dry. Finally, a GlOx layer was formed on the poly-l-lysine/poly(4-styrenesulfonate)-complex layer by crosslinking the enzyme by the addition of a glutaraldehyde solution. The use of thin bilayer system with the inner, polyion complex membrane, which showed permselectivity based on the solute size with the molecular cut-off of ≈100, brought high performance characteristics to the l-glutamate-sensing electrode; it showed high sensitivity (detection limit, 20 nM), rapid response (100% response time, 3 s), low interferential level (the ratio of response for l-ascorbic acid to that for the same concentration of l-glutamic acid, 8×10−2), and high stability (usable for more than a month). The bilayer-based electrode was useful for the rapid measurement of glutamate–oxaloacetate transaminase (GOT) and glutamate–pyruvate transaminase (GPT) in serum sample: each transaminase (0.2–1000 U l−1) could be determined within 10 s.