Sensitive detection of alkaline phosphatase by switching on gold nanoclusters fluorescence quenched by pyridoxal phosphate

被引：123

作者：

Halawa, Mohamed Ibrahim ^{[1
,2
,3
]}

Gao, Wenyue ^{[1
,2
]}

Saqib, Muhammad ^{[1
,2
]}

Kitte, Shimeles Addisu ^{[1
,2
]}

Wu, Fengxia ^{[1
,2
]}

Xu, Guobao ^{[1
]}

机构：

[1] Chinese Acad Sci, State Key Lab Elect Chem, Changchun Inst Appl Chem, Changchun 130022, Jilin, Peoples R China

[2] Univ Chinese Acad Sci, Beijing 100049, Peoples R China

[3] Univ Mansoura, Dept Pharmaceut Analyt Chem, Fac Pharm, Mansoura 35516, Egypt

来源：

BIOSENSORS & BIOELECTRONICS | 2017年 / 95卷

基金：

中国国家自然科学基金;

关键词：

Bovine serum albumin protected gold; nanoclusters; Pyridoxal phosphate; Alkaline phosphatase; Pyridoxal; Quenching; STABILIZED AU NANOCLUSTERS; CARBON QUANTUM DOTS; BIOLOGICAL APPLICATIONS; CASCADE REACTION; ENERGY-TRANSFER; SERUM-ALBUMIN; AT-BSA; ASSAY; NANOPARTICLES; BINDING;

D O I：

10.1016/j.bios.2017.03.073

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

In this work, we designed highly sensitive and selective luminescent detection method for alkaline phosphatase using bovine serum albumin functionalized gold nanoclusters (BSA-AuNCs) as the nanosensor probe and pyridoxal phosphate as the substrate of alkaline phosphatase. We found that pyridoxal phosphate can quench the fluorescence of BSA-AuNCs and pyridoxal has little effect on the fluorescence of BSA-AuNCs. The proposed mechanism of fluorescence quenching by PLP was explored on the basis of data obtained from high-resolution transmission electron microscopy (HRTEM), dynamic light scattering (DLS), UV-vis spectrophotometry, fluorescence spectroscopy, fluorescence decay time measurements and circular dichroism (CD) spectroscopy. Alkaline phosphatase catalyzes the hydrolysis of pyridoxal phosphate to generate pyridoxal, restoring the fluorescence of BSA-AuNCs. Therefore, a recovery type approach has been developed for the sensitive detection of alkaline phosphatase in the range of 1.0-200.0 U/L (R-2 =0.995) with a detection limit of 0.05 U/L. The proposed sensor exhibit excellent selectivity among various enzymes, such as glucose oxidase, lysozyme, trypsin, papain, and pepsin. The present switch-on fluorescence sensing strategy for alkaline phosphatase was successfully applied in human serum plasma with good recoveries (100.60-104.46%), revealing that this nanosensor probe is a promising tool for ALP detection.

引用

页码：8 / 14

页数：7

共 48 条

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