Time-Resolved Fluorescence Detection of Superoxide Anions Based on an Enzyme-Integrated Lanthanide Coordination Polymer Composite

Abstract

In this work, we proposed a new strategy of fabricating time-resolved fluorescent nanoprobes by using an enzyme-integrated lanthanide coordination polymer (CP) composite for the detection of superoxide anions (O₂^•-). This CP composite was constructed with terbium ions (Tb³⁺) as a metal node, adenosine triphosphate (ATP) as a bridge ligand, and carboxyphenylboronic acid (CPBA) as a sensitizer in which superoxide dismutase (SOD) was encapsulated by a self-adaptive inclusion process. The as-prepared SOD@ATP/Tb-CPBA displays both catalytic and fluorescence properties. Benefiting from the shielding effect of ATP/Tb CP, greatly enhanced catalytic activity and stability against harsh environments can be obtained in the loaded SOD. Meanwhile, the loaded SOD can remove the water molecules on the coordination sphere of Tb³⁺, leading to a significant increase in the fluorescence intensity and lifetime of SOD@ATP/Tb-CPBA. However, upon the addition of O₂^•-, the fluorescence of SOD@ATP/Tb-CPBA was quenched significantly. This is because SOD can convert O₂^•- into H₂O₂ to induce the deboronation of CPBA, resulting in an intramolecular charge transfer process. On this basis, by taking advantage of Tb³⁺ in long lifetime emission, a time-resolved fluorescence method was developed for the detection of O₂^•-, and satisfactory results have been achieved in both buffered aqueous solutions and serum samples. We believe that the presented study will open up a new avenue to develop enzyme-involved fluorescent nanoprobes.