牛血清蛋白纳米金团簇基于荧光共振能量转移法灵敏检测尿酸

doi:10.3964/j.issn.1000-0593(2022)02-0483-07

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摘要：基于纳米金团簇（AuNCs）良好的光学稳定性、生物相容性和简单无毒的制备方法，开发了一种具有高度选择性、高灵敏度且可视化的尿酸（UA）传感器。使用牛血清白蛋白（BSA）作为模板合成了BSA-AuNCs。在尿酸氧化酶的催化下，UA产生化学计量的过氧化氢（H₂O₂），导致AuNCs的荧光猝灭。此外，发现BSA-AuNCs在该体系模拟过氧化物酶发挥酶活性，它可以催化产物H₂O₂将底物3,3’,5,5’-四甲基联苯胺（3,3’,5,5’-tetramethylbenzidine, TMB）氧化成ox-TMB，此时BSA-AuNCs的发射光谱与ox-TMB的吸收光谱重叠，发生了一种荧光共振能量转移（FRET）。BSA-AuNCs作为供体将激发能量转移到受体ox-TMB，使ox-TMB产生荧光，同时BSA-AuNCs的荧光强度明显低于单独存在时的强度，从而大大提高了UA检测的灵敏度。在最佳条件下，发现UA浓度在2～100 μmol·L^-1猝灭程度线性关系良好，线性方程为（F₀-F）/F₀=0.005 85c_UA+0.103 64，线性相关系数为0.995 4，UA的检出限为0.26 μmol·L^-1，远低于正常人体UA水平的最低限（90 μmol·L^-1），同时研究了血液样本中UA的加标回收，回收率在97.3%～104.7%，表明了该方法在临床血样UA的检测中具有很大的应用潜力，为进一步的临床分析提供了良好的理论依据和方法学指导。

关键词：纳米金团簇；尿酸；过氧化氢；荧光共振能量转移

Abstract：Based on gold nanoclusters （AuNCs） with good optical stability, biocompatibility and simple and non-toxic preparation method, this paper developed a highly selective, highly sensitive and visualized uric acid （UA） sensor. We synthesized BSA-AuNCs using bovine serum albumin （BSA） as a template. Under the catalysis of urate oxidase, UA produces stoichiometric hydrogen peroxide （H₂O₂）, which causes the fluorescence of AuNCs to be quenched. In addition, we found that BSA-AuNCs mimic the peroxidase activity in this system, catalysing the substrate’s oxidation 3,3’,5,5’-tetramethylbenzidine （TMB） to ox-TMB by H₂O₂. At this time, the emission spectrum of BSA-AuNCs overlaps the absorption spectrum of ox-TMB, and a kind of fluorescence resonance energy transfer （FRET） occurs. BSA-AuNCs acts as a donor to transfer the excitation energy to the acceptor ox-TMB, which makes ox-TMB produce fluorescence. At the same time, the fluorescence intensity of BSA-AuNCs is significantly lower than when it exists alone, which significantly improves the sensitivity of UA detection. Under optimum conditions, it is found that the UA concentration has an excellent linear relationship between the quenching degree of 2～100 μmol·L^-1, the linear equation is （F₀-F）/F₀=0.005 85c_UA+0.103 64, the linear correlation coefficient is 0.995 4, and the detection limit is 0.26 μmol·L^-1, which is far below the minimum limit of normal human UA levels （90 μmol·L^-1）. Meanwhile, the recovery of UA in blood samples was studied, and the recovery rate was 97.3% to 104.7%, indicating that this method is effective in clinical blood samples. The detection has tremendous application potential, and provides an excellent theoretical basis and methodological guidance for further clinical analysis.

Key words：Gold nanoclusters; Uric acid; Hydrogen peroxide; Fluorescence resonance energy transfer

收稿日期: 2021-01-24 修订日期: 2021-03-20

中图分类号:

O657.3

基金资助: 国家重点研发计划项目（2016YFF0203802），北京市科技计划项目（Z201100009319001）资助

通讯作者: 罗云敬 E-mail: luoyj@bjut.edu.cn

作者简介: 丛剑涵，1993年生，北京工业大学环境与生命学部，环境与病毒肿瘤学北京市重点实验室硕士研究生
e-mail: 15732636263@163.com

引用本文:

丛剑涵，罗云敬，齐小花，邹明强，孔陈晨. 牛血清蛋白纳米金团簇基于荧光共振能量转移法灵敏检测尿酸[J]. 光谱学与光谱分析, 2022, 42(02): 483-489.
CONG Jian-han, LUO Yun-jing, QI Xiao-hua, ZOU Ming-qiang, KONG Chen-chen. Sensitive Detection of Uric Acid Based on BSA Gold Nanoclusters by Fluorescence Energy Resonance Transfer. SPECTROSCOPY AND SPECTRAL ANALYSIS, 2022, 42(02): 483-489.

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