Epitope Mapping of Monoclonal Antibodies

Purified MAbs 1A7 and 4E2 were selected for epitope mapping under three rounds of biopanning with the Ph.D-12 TM Phage Display Peptide Library Kit (New England BioLabs, Cambridge, MA, USA), as previously described [33 (link)]. Briefly, 96-well plates were coated with purified MAbs and incubated with blocking buffer. The phage library was then added to the plate and incubated for 1 h. After five washes with TBS buffer, 1 M Tris-HCl was added to the plate to elute the bound phages. 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 titer of the input phages.