仿生酶固定化纳米反应器通过级联催化反应实现胆固醇的精确检测  

Bio-inspired enzyme-immobilized nanoreactors with cascade catalytic reaction for accurate detection of cholesterol

作　　者：熊京 商灵灵 赵德偲 王丹 Jing Xiong;Lingling Shang;Decai Zhao;Dan Wang(State Key Laboratory of Biochemical Engineering,Key Laboratory of Biopharmaceutical Preparation and Delivery,Institute of Process Engineering,Chinese Academy of Sciences,1 North 2nd Street,Zhongguancun,Beijing 100190,China;University of Chinese Academy of Sciences,19A Yuquan Road,Beijing 100049,China)

机构地区：[1]State Key Laboratory of Biochemical Engineering,Key Laboratory of Biopharmaceutical Preparation and Delivery,Institute of Process Engineering,Chinese Academy of Sciences,1 North 2nd Street,Zhongguancun,Beijing 100190,China [2]University of Chinese Academy of Sciences,19A Yuquan Road,Beijing 100049,China

出　　处：《Science China Materials》2024年第8期2540-2549,共10页中国科学（材料科学）（英文版）

基　　金：supported by National Natural Science Foundation of China(52202354,52272097,52072369);Open Funding Project of the State Key Laboratory of Biochemical Engineering&Key Laboratory of Biopharmaceutical Preparation and Delivery(2023KF-04)。

摘　　要：基于酶的生化检测方法具有准确性高、检测速度快等优势,但生物酶的不稳定性限制了这一方法的应用.因此,开发一种活性高、稳定性强、反应效率高的酶包封体系是提高胆固醇生化检测的关键.为此,本研究发展了由二棕榈酸磷脂酰胆碱包覆中空多壳层结构的仿生人造细胞,HoMS的区室化结构和磷脂膜的选择透过性实现了酶的稳定包封和信号的高效传输.本研究通过信号的级联放大实现了胆固醇的快速准确检测.即利用负载胆固醇氧化酶的HoMS@DPPC引发胆固醇氧化反应,产生的中间产物H;2;O;2;作为信号分子,在负载辣根过氧化物酶的HoMS@DPPC催化作用下,促进荧光红染料生成试卤灵,再进行比色检测胆固醇的含量.由级联催化介导的胆固醇信号放大使得该系统可以应用于胆固醇检测,并且该方法表现出高的选择性和准确性,检测范围为0.05~0.3mM,检测限可达4.9μM.该方法可用于动物血清中胆固醇的检测,回收率为90.8%~104.9%,检测值与商业检测方法的结果高度吻合.此外,该系统可与智能设备集成,构建智能比色平台,实现图像分析与胆固醇浓度检测的一体化,为准确、快速的生化检测开辟了新途径.Although enzyme-based biochemical detection offers high accuracy and speed,their applications are limited by the instability of biological enzymes.Developing an enzyme-encapsulated system with high activity,strong stability,and high reaction efficiency is a crucial strategy for cholesterol detection.Herein,we developed a bionic artificial cell with hollow multishelled structures(HoMS)covered by phospholipid membranes.The compartmentalized structure and selective permeability of this artificial cell enable stable enzyme encapsulation and efficient signal transmission,achieving rapid and accurate cholesterol detection through signal cascade amplification.Cholesterol oxidase-loaded HoMS@DPPC was used to initiate cholesterol oxidation reaction,and the resulting intermediate product H2O2 was regarded as signal molecules to enter horseradish peroxidase-loaded HoMS@DPPC,promoting Amplex Red to produce resorufin.The amplification of cholesterol signal mediated by cascade catalysis makes the present system applicable to the detection of cholesterol,and the method shows high selectivity and accuracy,with a detection range of 0.05–0.3 mM and a detection limit of 4.9μM.It can be used for the detection of cholesterol in animal serum with a recovery of 90.8%to 104.9%,highly consistent with the results of commercial detection methods.Furthermore,the system can be integrated with smart devices to build an intelligent colorimetric platform,enabling the integration of image analysis and cholesterol concentration detection,opening up a new path for accurate and rapid biochemical detection.

关 键 词：图像分析 设备集成 生化检测 胆固醇氧化酶 选择透过性 信号放大 动物血清 酶固定化 

分 类 号：Q814[生物学—生物工程] O657.3[理学—分析化学] O643.3[理学—化学]