Binding of anti-GPIb mAbs to peptides displayed on phage proteins after SDS-denaturation and Western blotting.

mAb 27A10 (1, 2), mAb 12G1 (3, 4), mAb 12E4 (5, 6), mAb 6B4/clone D3 from the C7 library (7, 8), mAb 6B4/clone E1 from L15 library (9,10). Phages (2 × 1010) were run on SDS-PAGE (10% gel) either under nonreducing (lanes 1, 3, 5, 7, 9) or reducing (lanes 2, 4, 6, 8, 10) conditions, transferred to nitrocellulose membranes, and next probed with respective bmAbs. Detection occurred after incubation with streptavidin-HRP using enhanced chemiluminescence. mAbs 27A10, 12E4, and 12G1 recognized their phages under nonreducing, but not under reducing, conditions except for mAb 12E4. All phages selected on mAb 6B4 could be detected both under nonreducing and reducing conditions. Molecular weight (MW) marker is indicated.