A ProteOn™ XPR36 Protein Interaction Array System (Bio-Rad) was
used to measure the affinity of the monoclonal antibody 14-25-9 to His-tagged
recombinant PCNA. For the assay, a HTG sensor chip (#1765031) and ProteOn
Manager Version 3.1.0.6 (Bio-Rad Laboratories) were employed. After activation
of the chip using EDC/S-NHS amine coupling procedure, the ligand immobilization
process was performed with His-tagged recombinant PCNA and His-tagged IL2 as
control, at a flow rate of 30μl/min in different flow cells. Different
analyte (14-25-9) concentrations (0–100nM) were injected at a flow rate
of 40μl/min, with regeneration of the surface using 50mM NaOH after each
analyte. Data was analyzed using the bivalent binding model (40 (link)).
used to measure the affinity of the monoclonal antibody 14-25-9 to His-tagged
recombinant PCNA. For the assay, a HTG sensor chip (#1765031) and ProteOn
Manager Version 3.1.0.6 (Bio-Rad Laboratories) were employed. After activation
of the chip using EDC/S-NHS amine coupling procedure, the ligand immobilization
process was performed with His-tagged recombinant PCNA and His-tagged IL2 as
control, at a flow rate of 30μl/min in different flow cells. Different
analyte (14-25-9) concentrations (0–100nM) were injected at a flow rate
of 40μl/min, with regeneration of the surface using 50mM NaOH after each
analyte. Data was analyzed using the bivalent binding model (40 (link)).
