基于金属有机框架的固定化酶微反应器与毛细管电色谱集成用于高效酶测定,Analytical Chemistry

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基于金属有机框架的固定化酶微反应器与毛细管电色谱集成用于高效酶测定
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-04-23 , DOI: 10.1021/acs.analchem.1c05586
Rui Liu ₁ , Gaoyi Yi ₂ , Baian Ji ₃ , Xiuqiong Liu ₂ , Yuanqi Gui ₃ , Zhining Xia _{1,

4} , Qifeng Fu ₃

Affiliation

酶测定对于研究酶介导的生化反应以及临床诊断和药物开发非常重要。与毛细管电泳 (CE) 相结合的固定化酶微反应器 (IMER) 技术已经常用于在线酶测定。然而，传统的 IMER-CE 酶分析方法存在载量低、稳定性差等缺陷。本文首先探索了在酶固定化和毛细管电色谱（CEC）分离领域具有巨大潜力的金属有机骨架（MOFs）作为具有良好酶固定性能的新型载体材料和具有优异分离能力的固定相，以构建一种新型载体材料。用于高效在线酶测定的集成 MOFs-IMER-CEC 微量分析系统。₂纳米晶体涂层在毛细管内表面上具有丰富的晶间介孔率，并被用作 CEC 集成在线酶分析的高效和稳健的 IMER。_{UiO-66-NH 2}改性毛细管的优异分离性能通过中性、酸性和碱性三种化合物的高效分离得到验证。_{UiO-66-NH 2}的米氏常数和酶抑制动力学–IMER经过系统评估，与传统共价键合方法制备的CE集成IMER相比，具有显着提高的酶负载能力、对底物的亲和力以及显着提高的稳定性和可重复性等明显优势。此外，该方法成功用于豆科蔬菜样品中有机磷农药的检测，具有很强的实用性。该研究不仅为基于微通道的高性能IMER提出了一种新的支撑材料和构建策略，而且可以广泛应用于生物分析和生物传感研究。

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Metal–Organic Frameworks-Based Immobilized Enzyme Microreactors Integrated with Capillary Electrochromatography for High-Efficiency Enzyme Assay

Enzyme assays are important for studying enzyme-mediated biochemical reactions and for clinical diagnosis and drug development. The technique of an immobilized enzyme microreactor (IMER) integrated with capillary electrophoresis (CE) has been frequently utilized in online enzyme assays. However, the traditional approaches for IMER-CE enzyme analysis have some defects such as low loading capacity and poor stability. Herein, metal–organic frameworks (MOFs), which have enormous potential in the fields of enzyme immobilization and capillary electrochromatographic (CEC) separation, were first explored as novel support materials with good enzyme immobilization performance and stationary phases with excellent separation abilities to construct an integrated MOFs-IMER-CEC microanalysis system for a high-efficiency online enzyme assay. As a proof-of-concept demonstration, acetylcholinesterase (AChE) was immobilized on a densely packed UiO-66-NH₂ nanocrystal coating on a capillary inner surface with abundant intercrystalline mesoporosity and was employed as a highly effective and robust IMER for CEC-integrated online enzyme analysis. The excellent separation performance of the UiO-66-NH₂-modified capillary was verified by high-efficiency separation of three types of neutral, acidic, and basic compounds. The Michaelis–Menten constant and enzyme inhibition kinetics of UiO-66-NH₂–IMER were systematically assessed, exhibiting distinct advantages such as remarkably increased enzyme loadability, superior affinity for substrates, and greatly improved stability and repeatability compared to CE-integrated IMERs prepared by the traditional covalent bonding method. Furthermore, the developed method was successfully utilized for detecting organophosphorus pesticides in leguminous vegetable samples, demonstrating its strong practicality. The study not only proposed a novel support material and construction strategy for a high-performance microchannel-based IMER but also can be widely used in bioanalysis and biosensing research.

更新日期：2022-04-23

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