Label-Free, High-Throughput, Sensitive, and Logical Analysis Using Biomimetic Array Based on Stable Luminescent Copper Nanoclusters and Entropy-Driven Nanomachine

The development of an array for high-throughputand logical analysisof biomarkers is significant for disease diagnosis. DNA-templatedcopper nanoclusters (CuNCs) have a strong potential to serve as alabel-free photoluminescence source in array platforms, but theirluminescent stability and sensitivity need to be improved. Herein,we report a facile, sensitive, and robust biomimetic array assay byintegrating with stable luminescent CuNCs and entropy-driven nanomachine(EDN). In this strategy, the luminescent stability of CuNCs was improvedby adding fructose in CuNCs synthesis to offer a reliable label-freesignal. Meanwhile, the DNA template for CuNCs synthesis was introducedinto EDN with excellent signal amplification ability, in which thereaction triggered by target miRNA would cause the blunt/protrudingconformation change of 3 & PRIME;-terminus accompanied by the productionor loss of luminescence. In addition, a biomimetic array fabricatedby photonic crystals (PCs) physically enhanced the emitted luminescentsignal of CuNCs and achieved high-throughput signal readout by a microplatereader. The proposed assay can isothermally detect as low as 4.5pM of miR-21. Moreover, the logical EDN was constructed to achievelogical analysis of multiple miRNAs by AND or ORlogic gate operation. Therefore, the proposed assay has the advantagesof label-free property, high sensitivity, flexible design, and high-throughputanalysis, which provides ideas for developing a new generation offacile and smart platforms in the fields of biological analysis andclinical application.

Automated summary

This study presents a facile, sensitive, and robust biomimetic array assay by integrating luminescent copper nanoclusters (CuNCs) and entropy-driven nanomachine (EDN). By adding fructose in CuNCs synthesis, the luminescent stability of CuNCs was improved. The DNA template for CuNCs synthesis was introduced into EDN to achieve signal amplification and logical analysis of multiple miRNAs. The proposed assay showed label-free property, high sensitivity, flexible design, and high-throughput analysis, providing ideas for developing new platforms in biological analysis and clinical application.