The test-line and conjugate-line latex beads were sonicated and diluted in 1-day aged casein with 10% sucrose and 2% trehalose to a final concentration of 0.06% test-line beads and 0.015% control-line beads. The bead solution was sprayed in three discreet locations on the conjugate pad, 10, 15 and 20 mm from the upstream edge of the nitrocellulose. The conjugate was sprayed at 4 µL/cm using the Biodot Airjet (ZX1010, Biodot, Irvine, CA). Biotinylated MM08 was similarly diluted in 1-day aged casein with 10% sucrose and 2% trehalose to a final concentration of 75 µg/mL. It was striped using the Biodot Frontline 5mm from the upstream edge at 4 µL/cm.