Aptamer-functionalized magnetic nanoparticles conjugated organic framework for immobilization of acetylcholinesterase and its application in inhibitors screening.
Aptamer-functionalized magnetic nanoparticles conjugated organic framework for immobilization of acetylcholinesterase and its application in inhibitors screening.
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DOI：
10.1016/j.aca.2020.10.024
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发表时间：
2020-10
影响因子：
6.2
通讯作者：
Liping Zhao;Ge Yang;Linsen Li;Chao Zhu;Yao Ma;Feng Qu
中科院分区：
文献类型：
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作者：
Liping Zhao;Ge Yang;Linsen Li;Chao Zhu;Yao Ma;Feng Qu
基于适体功能化磁性纳米颗粒共轭有机框架（COF）的新型酶反应器的构建是本研究的重要内容之一。采用毛细管电泳法对乙酰胆碱酯酶适体进行了一轮筛选。首先合成了一种新型的富含羧基的磁性碳纳米管材料，然后用所选择的适体通过共价连接对其表面进行修饰。利用乙酰胆碱酯酶结合适体的高亲和力和特异性，固定化乙酰胆碱酯酶制备酶反应器Fe3O4@COF-Apt-AChE。对所构建的酶反应器进行了综合表征，并对影响其催化效率的关键因素进行了详细研究。利用适体对磁性COF材料进行表面修饰，使乙酰胆碱酯酶固定化，从而提高了固定化酶对底物的亲和力。具有良好的可重复使用性（8次以上）和较长的稳定性（42天后酶活性仍保持在90%）。最后，该酶反应器可用于AChE抑制剂的筛选，拓展了其应用能力。该方案不仅为新型核酸适配体功能化磁性COF材料作为酶反应器的制备开辟了新途径，而且还为核酸适配体与酶的整合提供了广阔的应用前景。
Construction of new enzyme reactor based on aptamer functionalized magnetic nanoparticles conjugated organic framework (COF) for acetylcholinesterase immobilization has been an enabling endeavor in this work. The aptamer against acetylcholinesterase was selected through a method based on capillary electrophoresis in one round. A new magnetic COF material rich of carboxyl groups was firstly synthesized, and its surface was then modified with the selected aptamer through covalently linking. Acetylcholinesterase was immobilized to fabricate the enzyme reactor Fe3O4@COF-Apt-AChE through the high affinity and specificity with its binding aptamer. The as constructed enzyme reactor was comprehensively characterized and the key factors that affected its catalysis efficiency were investigated in detail. Owing to the surface modification of the magnetic COF materials by aptamer for acetylcholinesterase immobilization, the immobilized enzyme exhibited improved substrates affinity. What’s more, good reusability (more than 8 times) and prolonged stability (enzyme activity still kept at 90% after 42 days) were also achieved. Finally, the enzyme reactor could be applied in AChE inhibitors screening, which expanded its application capability. The proposed protocol not only paves a new way for fabrication of novel aptamer functionalized magnetic COF materials as enzyme reactors, but also indicates a broadened application of the integration of aptamer and its enzyme.