Isolation and specificity of human P-selectin–binding phage.
(A) Selection on immobilized human PS-IgG using the pComb8 phage display peptide library, resulting in a 200-fold enrichment of P-selectin–binding phage in the fifth round of selection. (B) The enriched phage pools were specific for PS-IgG compared with human E-selectin–Ig (E-Sel-IgG) and L-selectin–IgG, and mouse PS-IgG (mP). Binding was reduced in the absence of calcium. Nonspecific phage pools (NS) showed little binding to human P-selectin. (C) Selected phage pools (Selection) specifically bound human P-selectin–transfected CHO cells compared with nonselected library phage (Library). Values are expressed as the ratio of acid-eluted phage (output) and input titer (generally 109-1010colony-forming units) and represent means of duplicate experiments ± variation.
