Hierarchical manganese dioxide nanoflowers enable accurate ratiometric fluorescence enzyme-linked immunosorbent assay

We first developed a green, mild and rapid method for the preparation of hierarchical manganese dioxide nanoflowers (MnO2 NFs) as nanozymes with intrinsic oxidase-like activity using citric acid for the reduction of potassium permanganate. The open structure of MnO2 NFs can lead to a larger specific surface area for improving the antibody loading amount and oxidase mimicking activity. In addition, carboxyl groups from the residual citric acid on the surface of MnO2 NFs afford a good affinity with the antibody through an amidation coupling reaction, which does not need complex surface modification anymore. Then, a reliable ratiometric fluorescence nanozyme-linked immunosorbent assay towards C-reactive protein (CRP) was designed successfully based on the MnO2 NFs. The high sensitivity and clinical feasibility of the presented methodology were demonstrated, which hold great promise in biomedical fields. Moreover, the mild and simple process for the synthesis of MnO2 NFs will make good contributions to many other applications.