Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, Ministry of Education, China Agricultural University, Beijing, 100193, China.
Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, Ministry of Education, China Agricultural University, Beijing, 100193, China; Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China.
Talanta. 2024 May 1;271:125664. doi: 10.1016/j.talanta.2024.125664. Epub 2024 Jan 12.
We presented a label-free fluorescent biosensor based on magnetic dual-aptamer allosteric regulation of β-lactoglobulin (β-LG) detection. The bovine serum albumin (BSA) acted as the bridge to connect amino-modified magnetic beads and aptamer, which synthesized pyramid-type probes (MBAP) with high capture and reduced nonspecific adsorption. Moreover, the original aptamer was tailored and then designed as a bivalent aptamer to fabricate allosteric signal probes (ASP). The ASP can both specifically capture β-LG and output the fluorescence signal. The detection mechanism is as follows. The combination of the dual-aptamer and β-LG triggered the allosteric change, resulting in the release of SYBR Green (SG I) from the allosteric signal probe and change signals. This method exhibits a broad linear detection range from 10 ng/mL to 1 mg/mL and the limit of detection reaches as low as 8.06 ng/mL. This study provides a highly generalizable strategy for protein biomolecular detection via replacing different target aptamers.
我们提出了一种基于磁性双适体对β-乳球蛋白（β-LG）检测的无标记荧光生物传感器。牛血清白蛋白（BSA）作为连接氨基修饰的磁性珠和适体的桥梁，合成了具有高捕获能力和减少非特异性吸附的金字塔型探针（MBAP）。此外，原始适体经过修饰后被设计成双价适体来制备变构信号探针（ASP）。ASP 既能特异性地捕获β-LG，又能输出荧光信号。检测机制如下。双适体与β-LG 的结合触发了变构变化，导致变构信号探针中的 SYBR Green（SG I）释放并改变信号。该方法在 10 ng/mL 至 1 mg/mL 的宽线性检测范围内表现出良好的线性关系，检测限低至 8.06 ng/mL。这项研究通过替换不同的靶适体，为蛋白质生物分子检测提供了一种高度通用的策略。
