Figure 2. Binding of TNP-specific fLC’s to immobilized TNP-OVA.

TNP-OVA was coated to the surface of 96-well plates. Unbound TNP-OVA was washed away followed by blocking with PBS/1%BSA. TNP-specific Ig-fLC was added and binding of Ig-fLC was detected using anti-mouse kappa light chain Ab conjugated with HRP. Results represent 6 individual experiments with duplicate binding curves. Binding curves were analyzed with non-linear fitting (Graphpad Prism 5.04): Kd: 76.9±3.8 nM; R²∶0.9905±0.0026; N = 12.