Breaking the Sensitivity-Specificity Trade-Off: A Nanoconfined Dual-DNA Aptamer Synergy Strategy

With the rising prevalence of vascular diseases, ultrasensitive and highly specific related protein monitoring has become crucial. However, traditional single-aptamer biosensors struggle to achieve an optimal balance between sensitivity and specificity, limiting their effectiveness in complex biosystems. Herein, we propose a dual-DNA aptamer synergy strategy with nanoconfined effect to overcome these challenges. Nanoconfined signal amplification significantly enhances the sensitivity of the dual-aptamer nanoconfined biosensor (N_CB_S), achieving a limit of detection (LOD) as low as 0.21 fM, while the synergistic effect of dual-aptamer confinement reduces the binding affinity (<i>K</i>_d), further improving protein recognition specificity. Moreover, it successfully performed multivariate discriminative analysis with 97% accuracy in clinical samples, with errors only occurring between different samples of the same disease. This synergistic dual-aptamer N_CB_S effectively balances sensitivity and specificity in protein recognition, promising high-precision biomarkers monitoring and advancing personalized medicine.