Magnetic separation-assisted cluster-amplified versatile fluorescent aptasensors for the sensitive detection of target biomolecules

Abstract

A sensitive and versatile platform for detecting diverse target biomolecules was developed by combining a magnetic separation module and a fluorescence amplification module in a plug-and-play manner. The magnetic separation module was constructed using magnetic beads (MBs), whose surfaces were modified with aptamer-blocked captor DNAs. The fluorescence amplification module was constructed by loading the fluorescent dye rhodamine 6G (Rh6G) into the pores of mesoporous silica nanoparticles (MSNs). The MSN surfaces were modified with prey DNAs, of which the MSN-near ends hybridized with complementary DNAs (sealing DNAs) to form duplexes to seal the pores, and the free ends were designed to be single-stranded that were complementary to the captor DNAs. Upon binding of targets to their aptamers, the captor DNAs were unblocked and thus were able to hybridize with the prey DNAs, to capture Rh6G-laden MSNs, forming MB-MSN clusters. The clusters were isolated by magnetic separation and heated to dissociate the DNA duplexes, to unseal the MSN pores and release the inner Rh6G; thus a target was converted into a cluster of Rh6G dyes. By simply changing the target aptamers and related DNA connectors, this strategy detected ATP, thrombin, and platelet-derived growth factor BB with detection limits of 2.1 nM, 4.1 pM, and 2.4 pM, respectively. A wide range of targets, high amplification efficiency and universal functional modules endow the aptasensors with good potential as versatile platforms for detecting target molecules in vitro and in medical research.