Extending the Operational Lifetime of Nucleic Acid-Based Electrochemical Sensors via Protection Against Competitive Displacement of Oligonucleotides
Abstract
Nucleic acid-based electrochemical sensors (NBEs) have emerged as a promising approach to continuous molecular monitoring in vivo. NBEs consist of electrically conducting gold surfaces coated with self-assembled monolayers of a mixture of electrode-passivating alkylthiols and functional alkylthiol-modified oligos. These oligos also display binding sites for the target analyte and redox reporters able to transfer electrons to the underlying gold electrode. Although sufficiently robust for continuous, multi-hour sensing of small molecules and proteins in biological fluids both in vitro and in vivo, NBEs decay over periods longer than 12 hours of continuous operation in these fluids. To address this issue, here we report a biofluid mimetic that can be leveraged to specifically study competitive displacement of oligonucleotides from NBEs, a critical sensor degradation pathway. Using this mimetic, we demonstrate three strategies that drastically mitigate competitive displacement and improve sensor stability in vitro. A combination of these strategies also improves sensor stability in vivo, demonstrated here via sensors emplaced in the brain cortex of live rats.
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