EGFR Binding Assay with MinE07UA-Biotin, MinE07UA'-Biotin, MinE07UB-Biotin, MinE07UC-Biotin and MinE07UC'-Biotin. The protein A plates were washed 3 times with 150 µL of wash buffer per well. EGFR-Fc protein was prepared in wash buffer at 1 µg/mL. 100 µL EGFR-Fc coating buffer per well was added and incubated for 30 minutes at 450 rpm at room temperature. The liquid was flicked out of the well plate. The plate was washed 3 times with wash buffer and then 50 µL 1x binding buffer to each well was added, and then incubate at 450 rpm for 10 minutes. MinE07UA-Biotin, MinE07UA'-Biotin, MinE07UB-Biotin, MinE07UC-Biotin and MinE07UC'-Biotin aptamers were prepared at concentrations 500 nM, 158 nM, 50 nM, 15.8 nM, 5 nM, 1.58 nM, 1 nM and 0.5 nM in binding buffer. The diluted aptamer was then denatured in the Eppendorf master cycler PCR machine (85°C for 5 minutes, then cooled to 25°C at 0.1°C per second and held at 25°C). The binding buffer was removed from the plate and washed 3 times with wash buffer. 100 µL of aptamer was added to the relevant wells. This was then incubated for 60 minutes at room temperature at 450 rpm. The liquid was flicked out of the well plate, and it was washed 3 times with wash buffer. Streptavidin-AP at a 1:1000 dilution in binding buffer was prepared, 100 µL was added to each well and it was incubated for 30 minutes at room temperature shaking at 450 rpm. The plate was then washed with wash buffer and then 95 µL of Tropix CDP-Star ready to use substrate was added and incubated for 10 minutes with shaking at 450rpm. The plate was then read again using the chemiluminescence protocol was then read on the Victor X4 plate reader. This was repeated until 3 biological repeats were achieved. Data analysis was done on Prism GraphPad Version 9.0.0 using the one site -specific binding model to calculated Bmax and K d .