School of Life Sciences, Pharmacy and Chemistry, Kingston University, Kingston upon Thames, London KT1 2EE, UK.
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Assiut University, Assiut 71526, Egypt.
Biosensors (Basel). 2024 Mar 20;14(3):151. doi: 10.3390/bios14030151.
Two fluorescent molecularly imprinted polymers (MIPs) were developed for pepsin enzyme utilising fluorescein and rhodamine b. The main difference between both dyes is the presence of two (diethylamino) groups in the structure of rhodamine b. Consequently, we wanted to investigate the effect of these functional groups on the selectivity and sensitivity of the resulting MIPs. Therefore, two silica-based MIPs for pepsin enzyme were developed using 3-aminopropyltriethoxysilane as a functional monomer and tetraethyl orthosilicate as a crosslinker to achieve a one-pot synthesis. Results of our study revealed that rhodamine b dyed MIPs (RMIPs) showed stronger binding, indicated by a higher binding capacity value of 256 mg g compared to 217 mg g for fluorescein dyed MIPs (FMIPs). Moreover, RMIPs showed superior sensitivity in the detection and quantitation of pepsin with a linear range from 0.28 to 42.85 µmol L and a limit of detection (LOD) as low as 0.11 µmol L. In contrast, FMIPs covered a narrower range from 0.71 to 35.71 µmol L, and the LOD value reached 0.34 µmol L, which is three times less sensitive than RMIPs. Finally, the developed FMIPs and RMIPs were applied to a separation-free quantification system for pepsin in saliva samples without interference from any cross-reactors.
两种荧光分子印迹聚合物（MIPs）被开发出来用于胃蛋白酶酶，使用荧光素和若丹明 B。两种染料的主要区别在于若丹明 B 的结构中存在两个（二乙氨基）基团。因此，我们希望研究这些官能团对所得 MIPs 的选择性和灵敏度的影响。因此，使用 3-氨丙基三乙氧基硅烷作为功能单体和正硅酸四乙酯作为交联剂，开发了两种基于二氧化硅的胃蛋白酶 MIPs，以实现一锅合成。我们的研究结果表明，与荧光素染色的 MIPs（FMIPs）相比，罗丹明 B 染色的 MIPs（RMIPs）具有更强的结合能力，其结合容量值为 256mg g，而 FMIPs 的结合容量值为 217mg g。此外，RMIPs 在胃蛋白酶的检测和定量方面表现出更高的灵敏度，线性范围从 0.28 到 42.85µmol L，检测限（LOD）低至 0.11µmol L。相比之下，FMIPs 的线性范围较窄，从 0.71 到 35.71µmol L，LOD 值达到 0.34µmol L，灵敏度比 RMIPs 低三倍。最后，所开发的 FMIPs 和 RMIPs 被应用于唾液样品中胃蛋白酶的无需任何交叉反应物干扰的无分离定量系统。
