We demonstrate continuous-flow biomolecule concentration and detection in a microfabricated slanted sieving structure, which we term a herringbone nanofilter array (HNA). The HNA structure consists of periodically-patterned deep and shallow nanoslits meeting at right angles. In addition to concentration, we can discriminate different sized analytes by mixing a fluorescent probe with the sample and measuring the extent of the concentrating effect. Using this principle, we interrogate biomolecular interactions, including protein–DNA binding, protein–protein interaction and antibody–antigen binding. The final example demonstrates a novel method to perform a homogeneous immunoassay for detecting a disease marker, human C-reactive protein (CRP), using fluorescent-labeled antibodies at clinically relevant concentrations. The signal amplification potential and continuous flow operation provide a significant advantage over other microfluidic batch separation techniques for the easy integration of this device into a common point-of-care diagnostic platform.
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