Table 1.

Epitope mapping and identification of the relative affinity for each Psl mAb

EpitopemAbKdPhage libraryCell attachment max. inhibitionOPK EC50
nMµg/mlµg/ml
Class 1Cam-003144.00Naive1.00.0220
Cam-0042,100.00Naive>30.00.2771
Cam-0058,400.00Naive>30.0>30.0
Class 2WapR-0010.84Patient30.00.3100
WapR-00312.20Patient30.00.2778
WapR-00212.60Patient30.00.3960
Class 3WapR-01675.00Patient10.00.2417
  • Epitope mapping was performed by competition ELISA and confirmed using an Octet flow system with Psl derived from the supernatant of an overnight culture of PAO1. The results indicate that Psl antibodies bind to at least three epitopes, which we refer to as class I, II, and III. Class I and II antibodies do not compete for binding; however, the class III antibody, WapR-016, partially inhibits binding of the class I and II antibodies. Antibody affinity was determined by Octet binding assays using Psl derived from the supernatant of overnight PAO1 cultures. Antibody KD was determined by averaging the binding kinetics of seven concentrations for each antibody. Also shown is a summary of cell attachment and OPK data experiments for each antibody. ELISA and antibody affinity was determined using three independent studies.