Effect of the nonspecific binding in differential impedance biosensing

The detection limits of impedance biosensors are dictated by the presence of background nonspecific binding, yet almost all the detection limits reported in the literature are determined using a clean buffer solution without confronting this real challenge. In this work, the authors employed the simplest "differential" impedance biosensor, composed of poly-l-lysine-polyethylene glycol-biotin-coated gold electrodes for the detection of streptavidin in the presence of 0.1% fetal calf serum, and studied the effect of the nonspecific binding on the performance of the differential impedance biosensing. The lowest streptavidin concentration detected by the system (5 μg/ml) was 1 order of magnitude higher (worse) than that from a previously demonstrated impedance biosensor where avidin was detected in the absence of background proteins. Interestingly, the origin of the differential signal was not due to the electrochemical properties of streptavidin itself but was that of the serum, where the coverage of the electrode by streptavidin indirectly modulated the electrical signal by suppressing the accessibility of the serum to the electrode.