聚环氧乙烷修饰的芘用于“荧光开”检测胰蛋白酶的研究

本文采用聚环氧乙烷(PEO)对疏水染料芘进行亲水化修饰设计,以获得具有良好水溶性和荧光性能的两亲性探针分子聚环氧乙烷修饰的芘(PEO-Py).以牛血清白蛋白(BSA)作为胰蛋白酶的水解底物以及猝灭剂,BSA加入会通过动态猝灭使荧光强度降低.加入胰蛋白酶水解后,荧光强度部分恢复,建立了基于荧光强度升高的“荧光开”胰蛋白酶检测方法.对胰蛋白酶的检测限可达到9.8 μg·mL-1,检测线性范围为0～0.075 mg·mL-1.其他蛋白不会引起荧光强度的降低,检测体系具有良好的选择性.通过亲水化修饰提高了芘衍生物的水溶性,建立了制备简单、成本低、水溶性好的检测探针,拓展了芘在生理环境中的检测应用.

Fluorescence "Turn on" Detection of Trypsin Using PEGylated Pyrene

The development of rapid,easily prepared and low-cost fluorescent probes for the determination of trypsin activity is still highly desired.PEGylated pyrene (PEO-Py) exhibited characteristic fluorescence properties and improved solubility in aqueous solution.BSA was designed specially to act as the substrate of trypsin and quencher.The fluorescence of PEO-Py was dynamic quenched upon the addition of BSA.Subsequent addition of trypsin into the system led to the cleavage of BSA into peptide fragments,and the fluorescence of PEO-Py was recovered to some extent.This phenomenon then allowal for the "turn on" detection of trypsin with a limit of detection of 9.8 μg·mL-1,and the linear range of trypsin was from 0 to 0.075 mg· mL-1.Other proteins didn't show obvious interference with the determination.Because of the simplicity,low cost and good solubility in aqueous buffer,PEO-py were believed to expand their applications in the analysis of biological samples.