Characterizing ATRXepi Binding to Immobilized PMab-1

Rat anti-MAP mAb PMab-1 (IgG_2a, κ) was described previously.^{(15 (link))} PMab-1 was immobilized on CM5 sensor chips using amino coupling chemistry according to the method provided by the manufacturer. The MAP-tagged ATRXepi proteins were diluted in PBST (phosphate-buffered saline, pH 7.4, containing 0.005% Tween 20) and injected at a flow rate of 30 μL/min. The ATRXepi was diluted in PBST (12.5, 25, 50, 100, and 200 nM) and passed over the biosensor chip. The binding was monitored for 60 seconds, followed by dissociation in running buffer for 120 seconds using the Single-cycle kinetics method. Binding curves were analyzed using BIAcore X100 Evaluation Software (GE Healthcare, Piscataway, NJ) with curve fitting using a 1:1 binding model.