酶活性触发的等离子体共振能量转移阻断用于酸性磷酸酶的高选择性检测。

Enzyme Activity Triggered Blocking of Plasmon Resonance Energy Transfer for Highly Selective Detection of Acid Phosphatase.

机构信息

Key Laboratory of Luminescent and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Chemistry and Chemical Engineering , Southwest University , Chongqing 400715 , China.

Non-linear Circuit and Intelligent Information Processing, College of Electronic and Information Engineering , Southwest University , Chongqing 400715 , China.

出版信息

Anal Chem. 2020 Jan 21;92(2):2130-2135. doi: 10.1021/acs.analchem.9b04685. Epub 2019 Dec 31.

DOI:10.1021/acs.analchem.9b04685

PMID:31850751

Abstract

Plasmon resonance energy transfer (PRET), as a new form of energy transfer first discovered in 2007, has been widely applied for the biomolecular recognition, detection of ions, cellular physiological status monitoring, and energy conversion. It occurs between noble metal nanoparticles (donor) and conjugated molecules or nanoparticles (acceptor). In this study, we used urchin-like gold nanoplasmonics (UGPs) and TMB as a new donor-acceptor pair to establish a novel PRET coupling system, avoiding trivial modification. PRET from UGPs to conjugated redox-active TMB leads to resonant quenching in the localized surface plasmon resonance (LSPR) spectra. However, when the acid phosphatase (ACP) was introduced, the hydrolyzate ascorbic acid (AA) converted from 2-phospho-l-ascorbic acid trisodium salt (AAP) could be capable of reducing TMB into TMB, thereby preventing the occurrence of PRET. The recovery of the scattering spectral intensity of UGPs was linearly related to the concentration of ACP in the range of 0.1 to 5.0 U/L, and the ACP with a detection limit of 0.076 U/L could be measured. In addition, this method also showed good selectivity attributed to the substrate specificity of enzyme.

摘要

等离子体激元共振能量转移（PRET）作为 2007 年首次发现的一种新的能量转移形式，已广泛应用于生物分子识别、离子检测、细胞生理状态监测和能量转换等领域。它发生在贵金属纳米粒子（供体）和共轭分子或纳米粒子（受体）之间。在本研究中，我们使用了海胆状金纳米等离子体（UGPs）和 TMB 作为新的供体-受体对，建立了一种新的 PRET 偶联系统，避免了繁琐的修饰。UGPs 到共轭氧化还原活性 TMB 的 PRET 导致局域表面等离子体共振（LSPR）光谱中的共振猝灭。然而，当引入酸性磷酸酶（ACP）时，2-磷酸-L-抗坏血酸钠盐（AAP）水解产生的抗坏血酸（AA）能够将 TMB 还原为 TMB，从而防止 PRET 的发生。UGPs 散射光谱强度的恢复与 0.1 至 5.0 U/L 范围内的 ACP 浓度呈线性关系，检测限为 0.076 U/L 的 ACP 可以被测量到。此外，该方法还表现出良好的选择性，这归因于酶的底物特异性。