Fluorescent Fingerprint Identification of Protein Structural Changes and Disease-Specific Amyloid Beta Aggregates Based on a Single-Nanozyme Sensor Array

The misfolding of amyloid β (Aβ) peptides into an aggregation state is a central hallmark of the onset of Alzheimer's disease (AD). However, conventional methods are mainly focused on detecting a specific Aβ peptide, which makes it difficult to recognize multiple analytes with different topological features and unfolded states at the same time. Here, we propose a simple and universal sensing strategy to construct a fluorescence sensor array by using a single-nanozyme probe combined with three fluorescent substrates as three recognition units to probe the protein structural changes and identify between multiple Aβ assemblies. In this sensor system, the fingerprint-like patterns are produced from the nonspecific interactions between topological proteins and the sensing units. As a result, this sensor array can accurately identify 13 kinds of proteins and their mixtures at different ratios. Moreover, the sensor array can discriminate against proteins with unfolded states and diverse conformational forms. Most importantly, the sensor array successfully distinguishes between multiple Aβ species, even in artificial cerebrospinal fluid samples and human serum samples. This work provides an attractive and reliable strategy for predicting pathologically relevant proteins and clinical diagnosis of AD.