基于羧基化单壁碳纳米角的荧光传感体系检测胰蛋白酶  

A fluorescence sensing system for the detection of trypsin based on carboxylic group-functionalized single-walled carbon nanohorns

作　　者：冯婷婷 闫姝竹 马洪志 杨丽霞 王晓华[2] FENG Tingting;YAN Shuzhu;MA Hongzhi;YANG Lixia;WANG Xiaohua(College of Traditional Chinese Medicine and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619,China;Pharmacy School of Guilin Medical University,Guilin 541004,China)

机构地区：[1]山西中医药大学中药与食品工程学院,晋中030619 [2]桂林医学院药学院,桂林541004

出　　处：《分析试验室》2024年第1期24-29,共6页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(82003925);山西中医药大学科技创新能力培育项目(2020PY-JC-10和2022PY-TH-24)资助。

摘　　要：构建了一种基于羧基化单壁碳纳米角(oxSWCNHs)的简单、灵敏的荧光传感体系用于胰蛋白酶的检测。实验中合成了水溶性的oxSWCNHs,并设计了一段羧基荧光素(FAM)标记肽段作为胰蛋白酶的底物,该底物能与胰蛋白酶进行特异性反应。当体系中没有胰蛋白酶存在时,FAM标记的肽段可以被oxSWCNHs强烈吸附,致使体系具有较低的荧光背景;当体系中有胰蛋白酶存在时,胰蛋白酶可以与其底物肽段发生特异性水解反应,导致FAM标记的肽段水解为单个氨基酸,致使FAM远离oxSWCNHs表面,FAM荧光不能被oxSWCNHs淬灭,体系具有较强的荧光强度,从而实现荧光传感体系对胰蛋白酶的检测。该传感体系线性范围为0~2.5μg/mL,可用于尿液中胰蛋白酶的测定。A simple fluorescent sensing system for the detection of trypsin by assembling a dye-labeled peptide with carboxylic group-functionalized single-walled carbon nanohorns(oxSWCNHs) was constructed.oxSWCNHs was prepared by nitric acid oxidation of single-walled carbon nanohorn,and the carboxyfluorescein(FAM) labeled peptide(FAM-RRRRRR) was chosen as a trypsin-sensitive core substrate,which can react specific with trypsin.Without the trypsin in the system,FAM-labeled peptide could be strongly adsorbed by oxSWCNHs,resulting in a low fluorescence background of the system.The system reacted with trypsin to cleave the peptide,resulting in the release of the dye moiety and a substantial increase in fluorescence intensity,and trypsin could be quantified accordingly.The linear range of the system was 0-2.5 μg/mL,and the method can be applied to detect trypsin in urine samples.

关 键 词：单壁碳纳米角 荧光传感体系 胰蛋白酶 

分 类 号：O657.39[理学—分析化学]