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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Controllable and high‐performance immobilized enzyme reactor: DNA‐directed immobilization of multienzyme in polyamidoamine dendrimer‐functionalized capillaries
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Controllable and high‐performance immobilized enzyme reactor: DNA‐directed immobilization of multienzyme in polyamidoamine dendrimer‐functionalized capillaries

机译:可控和高性能固定的酶反应器:DNA定向固定在聚酰胺胺树枝状细胞官能化毛细血管中的偏酶的固定

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Abstract In recent years, CE‐integrated immobilized enzyme reactors (IMERs) for single‐enzyme immobilization have attracted considerable attention. However, there has been little research on multienzyme immobilization in CE. Here, we introduce a method for fabricating a CE‐integrated IMER, using DNA‐directed immobilization to fix glucose oxidase and horseradish peroxidase in the capillary, which had been functionalized with polyamidoamine dendrimer (PAMAM). Owing to the reversibility of DNA hybridization, the reactor is capable of dynamic immobilization. Moreover, by introducing the PAMAM, the loading capacity of the IMER is greatly enhanced, and the PAMAM can spontaneously form complexes with DNA and then contribute to the efficiency and stability of the reactor. After 25 days storage, the prepared IMER ultimately retained approximately 70% of its initial activity. We also used the IMER to detect glucose, and the favorable linearity was obtained over the concentration range of 0.78–12.5?mM, with an LOD of 0.39?mM, demonstrating that the CE‐integrated IMER can be applied to actual samples. We believe that this strategy can be extended to other multienzyme immobilization systems, and CE‐integrated IMERs are potentially useful in a wide range of biochemical research applications.
机译:摘要:近年来,单酶固定化CE-集成固定化酶反应器(IMERs)已经吸引了相当多的关注。然而,一直在CE多酶固定化的研究很少。这里,我们介绍了用于制造CE-集成IMER,在毛细管,已被官能化聚酰胺 - 胺树枝状聚合物(PAMAM),使用DNA指导的固定到固定的葡萄糖氧化酶和辣根过氧化物酶的方法。由于DNA杂交的可逆性,该反应器能够动态固定的。此外,通过引入PAMAM,所述IMER的装载能力大大增强,并且可以PAMAM自发形成与DNA的复合物,然后向反应器的效率和稳定性。经过25天贮存,将制备的IMER最终保留其初始活性的约70%。我们还使用了IMER检测葡萄糖和获得良好的线性在0.78-12.5?毫的浓度范围内,以0.39 -3毫米的LOD,表明CE-集成IMER可以应用于实际样品。我们认为,这种策略可以扩展到其他的多酶固定系统,以及CE-集成IMERs是在广泛的生化研究应用可能有用。

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