Poly(ethylene glycol) hydrogel-encapsulated fluorophore-enzyme conjugates for direct detection of organophosphorus neurotoxins
Poly(ethylene glycol) hydrogel-encapsulated fluorophore-enzyme conjugates for direct detection of organophosphorus neurotoxins
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DOI：
10.1021/ac990901u
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发表时间：
1999-11-01
影响因子：
7.4
通讯作者：
Wild, JR
中科院分区：
文献类型：
--
作者：
Russell, RJ;Pishko, MV;Wild, JR
介绍了一种用包封的半荧光素(SNAFL)-有机磷水解酶偶联物制备聚乙二醇水凝胶材料的简单方法。直接测定酶催化的神经毒素水解是由自参考，ph敏感染料SNAFL-1提供的，其发射光谱在λ = 550响应ph变化。使用荧光光谱法和对氧磷作为模型有机磷，低至8 × 10(-7) M的对氧磷浓度可以很容易地检测到。根据信噪比确定检测限为16 nM。该材料在环境条件下储存时，具有较高的抗酶变性，浸出和光漂白的稳定性。
A simple approach is described for preparing poly(ethylene glycol) hydrogel materials with encapsulated seminapthofluorescein (SNAFL)-organophosphorus hydrolase enzyme conjugates. Direct determination of enzyme-catalyzed neurotoxin hydrolysis is provided by the self-referencing, pH-sensitive dye SNAFL-1, whose emission spectrum changes at lambda = 550 in response to pH. Using spectrofluorimetry and paraoxon as a model organophosphate, paraoxon concentrations as low as 8 x 10(-7) M could be readily detected. On the basis of the signal-to-noise ratio, a detection limit of 16 nM was determined. The materials demonstrated high stability against enzyme-denaturing, leaching, and photobleaching when stored under ambient conditions.