Epitope mapping with SPR showing sensorgrams of time (seconds) versus RU. (A) Sequences of peptides analyzed for epitope mapping of mAb-6B11 and mAb-1286. Amino acid sequences of full-length AP-FXIII and all overlapping peptides used for the epitope mapping are depicted. AP8 and AP9 differ in residue 34 containing either a Val or Leu, respectively. (B) Epitope mapping of mAb-6B11. Peptides AP1-AP9 were tested for binding to mAb-6B11 at a concentration of 10μM. All peptides are listed downward corresponding to their colored curves. (C-D) Affinity analysis of mAb-6B11 to AP1-12 and AP3-12. The affinity of mAb-6B11 to AP1-12 was measured at the following concentrations: 5 (green), 10 (cyan), 15 (dark blue), 20 (pink), and 30nM (brown) and to AP3-12 at the concentrations: 5 (green), 7.5 (aquamarine), 10 (pink), 15 (cyan), and 20nM (gray). (E) Localization of the binding site of mAb-6B11. mAb-6B11 was tested for binding to AP-FXIII-Val at a concentration of 16nM (purple) and AP13-37 at 150nM (pink). (F) Localization of the binding site of mAb-1286. The following peptides were tested for binding: AP-FXIII-Val at a concentration of 17.5nM (cyan), and AP13-37 (brown), and AP1-12 at 30nM (pink).