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基于牛血清白蛋白包覆金纳米簇过氧化物酶活性调控及金纳米双锥体蚀刻的胰蛋白酶检测多色生物传感器

Multicolor Biosensor for Trypsin Detection Based on the Regulation of the Peroxidase Activity of Bovine Serum Albumin-Coated Gold Nanoclusters and Etching of Gold Nanobipyramids.

作者信息

Luo Qin, Tian Mengjian, Luo Fang, Zhao Min, Lin Cuiying, Qiu Bin, Wang Jian, Lin Zhenyu

机构信息

Ministry of Education Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian350116, China.

Department of Gynecology and Obstetrics, The First Affiliated Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, Fujian361005, China.

出版信息

Anal Chem. 2023 Jan 31;95(4):2390-2397. doi: 10.1021/acs.analchem.2c04418. Epub 2023 Jan 13.

Abstract

The detection of trypsin is significantly important for both clinical diagnosis and disease treatment. In this study, an innovative multicolor sensor for trypsin detection has been established based on the regulation of the peroxidase activity of bovine serum albumin-coated gold nanoclusters (BSA-Au NCs) and efficient etching of gold nanobipyramids (Au NBPs). BSA-Au NCs have slight peroxidase enzyme activity and can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to generate TMB, while trypsin can hydrolyze BSA ligands on the surface of BSA-Au NCs, thus exposing more catalytic active sites of BSA-Au NCs and resulting in the enhancement of the peroxidase activity of BSA-Au NCs, hence more TMB is generated. Under acidic conditions, TMB can etch Au NBPs efficiently, consequently affecting the aspect ratio of Au NBPs accompanied by the ultraviolet-visible (UV-vis) spectra blue shifting of the system. Furthermore, this also results in color variations that can be distinguished and recognized by naked eyes without any expensive and sophisticated instruments. This multicolor sensor has an available linear relationship with the logarithm of the trypsin concentration in the range of 0.1-100 μg/mL, and the detection limit is 0.045 μg/mL. The designed sensor has been used to detect the concentration of trypsin in human serum samples from healthy individuals and pancreatitis patients with satisfactory results.

摘要

胰蛋白酶的检测对于临床诊断和疾病治疗都具有重要意义。在本研究中,基于牛血清白蛋白包覆的金纳米簇(BSA-Au NCs)过氧化物酶活性的调控以及金纳米双锥体(Au NBPs)的有效蚀刻,建立了一种用于胰蛋白酶检测的创新型多色传感器。BSA-Au NCs具有轻微的过氧化物酶活性,能够催化3,3',5,5'-四甲基联苯胺(TMB)氧化生成TMB,而胰蛋白酶可以水解BSA-Au NCs表面的BSA配体,从而暴露出更多BSA-Au NCs的催化活性位点,导致BSA-Au NCs的过氧化物酶活性增强,进而生成更多的TMB。在酸性条件下,TMB可以有效地蚀刻Au NBPs,从而影响Au NBPs的纵横比,同时系统的紫外可见(UV-vis)光谱发生蓝移。此外,这还会导致颜色变化,肉眼即可分辨和识别,无需任何昂贵且复杂的仪器。该多色传感器在0.1-100μg/mL范围内与胰蛋白酶浓度的对数具有良好的线性关系,检测限为0.045μg/mL。所设计的传感器已用于检测健康个体和胰腺炎患者血清样本中胰蛋白酶的浓度,结果令人满意。

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