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The regulation effect of surface charges on the transport of electrons in nanomaterials and ions in nanofluidic devices has been widely used to develop highly sensitive and label-free electrical biosensors. The intrinsic limitation to the clinical application of surface charge-effect nano-electrical biosensors is that they usually do not function in physiological conditions normally with high ionic concentrations (∼160 mM), in which the surface charges are screened within a short distance (&lt;1 nm at 160 mM). In this work, we developed a general strategy that enables surface charge-effect electrical biomolecular detection in physiological conditions with an integrated mechanism for enhancement of the limit of detection (LOD) by in situ preconcentration of target molecules during incubation and creation of a transient low ionic concentration environment during the signal read-out step using reconfigurable ion concentration polarization (ICP). We demonstrated the effectiveness of this strategy in a simple nanofluidic biosensor named a nanofluidic crystal (NFC), which can be prepared within hours and without expensive equipment. Our results indicate that the ion depletion effect of ICP could lower the ionic concentration by at least 200 fold and provide a stable ionic environment for over 15 s, enabling electrical detection of proteins and DNAs in serum and urine with LODs of 1-10 nM. We further reconfigured the device to preconcentrate target biomolecules before detection using the enrichment effect of ICP, obtaining LODs of 10-100 pM for proteins and DNAs in physiological conditions. By overcoming the inherent constraint on buffer conditions and the issues regarding fabrication, we believe that this work represents significant progress towards the practical application of surface charge-effect nano-electrical biosensors in point-of-care diagnostics and clinical medicine.

lab on a chip

Enabling electrical biomolecular detection in high ionic concentrations and enhancement of the detection limit thereof by coupling a nanofluidic crystal with reconfigurable ion concentration polarization