Abstract: Day 1: 1st round bio-panning 1.1Add 2 µM mouse IgG 1 (as mouse Fc blocker) to theHX02 human Fab phage library (Humanyx Pte Ltd) and incubate at RT, 30min. 1.2Wash 60 µl of DynaBeads M-280 Streptavidin (Invitrogen #11205D) with 1 ml PBS twice. 1.3Add 100 nM of biotinylated SARS-CoV-2 RBD-mFc protein in casein to the beads and incubate at room temperature for 1 hour. 1.4Wash beads 4 times with 1 ml PBS. 1.5Add the pre-mix of phage library and mouse IgG 1 to the washed beads of biotinylated SARS-CoV-2 RBD-mFc. 1.6Mix for 1 hour at room temperature. 1.7Wash beads 7 times with 1.5 ml 0.1% PBST. Bio-panning Round 1protocols.io| https://dx.doi.org/10.17504/protocols.io.bitykepw 2 1.8Add 250 µl of 0.1M TEA (Triethylamine) to beads at room temperature for 15 min to elute bound phage. 1.9Collect eluted phage using magnet and neutralize pH with 125 µl of 1M Tris pH 8. 1.10Infect neutralized phage to TG1 cells at 37°C for 45 min and spread phage infected cells onto 150 mm AG agar plates (2xYT agar, 100µg/ml Ampicillin, 2% Glucose). 1.11Incubate agar plates at 37°C overnight. 2Day 2: Harvest and package phages 2.1Harvest the phage infected TG1 cells by scraping up the cells on the agar plates and let it grow to OD 600nm ~0.5 at 37°C, shaking at 250rpm. 2.2Infect the bacteria culture with M13K07 helper phage (NEB #N0315S) at 37°C for 45 min. 2.3Culture the M13K07 helper phage infected cells at 30°C for 7 hrs with shaking at 250 rpm in AKG media (2x YT media, 100 µg/ml Ampicillin, 25 µg/ml Kanamycin).protocols.