Site-Specific Immobilization of ZNRF3 Reveals the Importance of Target Structural Integrity on Macrocyclic Peptide Selections

= 124 nM), emerging from the shared pool. This work reports a technique to selectively disrupt protein domains during phage selections, while also demonstrating that the structural integrity of immobilized targets is a primary determinant of phage display success. Not only does the necessity to maintain structural integrity influence sequence composition and affinities of peptides toward native protein targets, but also the overall enrichment efficiency of the selection itself. While disruptive immobilization may still yield useful ligands, strategies that preserve native folds enhance phage enrichment and maximize the identification of biologically relevant binders.