Epitope Mapping Using Phage Display

The epitope was mapped with purified 3B6 mAb using the Ph.D-12^TM Phage Display Peptide Library Kit (New England BioLabs), as previously described24 (link)25 (link). Three rounds of biopanning were performed. Briefly, each well of a 96-well plate was coated with 10 μg/mL of purified 3B6 mAb and incubated with blocking buffer. The phage library was then added to the plate and incubated for 1 hour. After five washes with TBS buffer, 1 M Tris-HCl was added to the plate to elute the bound phages24 (link)25 (link). The phages were then amplified and titred on LB/IPTG/Xgal plates for selection. The ratio of output to input was calculated as the titre of the amplified output phages to the titre of the input phages.