The nitrocellulose, 25 mm CN95 (Sartorius), was striped with a test line at 8 mm from the edge of (upstream from the flow direction) and 13 mm from the upstream edge of nitrocellulose. The test line was striped at 1 µL/cm with 1 mg/mL polystreptavidin (Cat #10 120 050, Biotez, Berlin, DE) and the control line was striped at 1 µL/cm 0.5 mg/mL goat anti-Chicken IgY (Cat #703-005-155, Jackson ImmunoResearch). Both lines were striping using a Biodot Frontline dispenser (XYZ3060, Biodot, Irvine, CA).