Figure 1
From: Molecular basis for epitope recognition by non-neutralizing anti-gp41 antibody F240
Kinetic and thermodynamic characterization of mAb F240 binding to gp41583–618 peptide.
(A) Surface Plasmon Resonance (SPR) analysis of mAb F240 binding to the gp41583–618 peptide (red lines) and the gp41583–618 peptide pretreated in reducing (with 0.05 nM TCEP in PBS) conditions (green lines). Sensorgrams were obtained at room temperature for mAb F240 immobilized on a Protein A chip with 0–200 nM concentrations of gp41 peptide passed over the chip. Black and red/green curves correspond to the experimental data (for concentrations of peptide in a range of 3.1–200 nM) and best fit using the BIAevaluation software, respectively. Standard deviations of ka, kd and KD for two experiments are shown. (B) Isothermal titration calorimetry (ITC) curves for mAb F240 binding to gp41583–618 (left) and gp41583–618 C698AC605A mutant (right), respectively. The binding isotherms were fitted to a one-site binding model using Origin for ITC version 7.0383 (MicroCal). The fitting yields the stoichiometry (N), the binding constant (KD), and the enthalpy (ΔH) of the binding reaction.
