A shear-enhanced CNT-assembly nanosensor platform for ultra-sensitive and selective protein detection

Detection and quantification of low-concentration proteins in heterogeneous media are generally plagued by two distinct obstacles: lack of sensitivity due to high dissociation equilibrium constant KD and non-specificity due to an abundance of non-targets with similar KD. Herein, we report a nanoscale protein-sensing platform with a non-equilibrium on-off switch that employs dielectrophoretic and hydrodynamic shear forces to overcome these thermodynamic limitations with irreversible kinetics. The detection sensitivity is achieved with complete association of the antibody-antigen-antibody (Ab-Ag-Ab) complex by precisely and rapidly assembling carbon nanotubes (CNT) across two parallel electrodes via sequential DC electrophoresis and AC dielectrophoresis (DEP), and with single-CNT electron tunneling conductance. The high selectivity is achieved with a critical hydrodynamic shear rate between the activated dissociation shear rates of target and non-target linkers of the aligned CNTs. We are able to reach detection limits of 100 attomolar (aM) and 10 femtomolar (fM) in pure samples for two ELISA assays with low and high dissociation constant: biotin/streptavidin (10 fM) and HER2/HER2 antibody (0.44 ± 0.07 nM), respectively. For both models, irreversible capture and shearing allow us to tune the dynamic range up to 5 decades by increasing the CNT numbers. We also demonstrate in spiked serum sample high selectivity towards target HER2 proteins against non-target HER2 isoform of a similar KD. The detection limit for HER2 in serum is lower than 100 fM.