To extend the capacity of GNP fluorescence quenching assay in probing the binding thermodynamics of high affinity MLGIs, we also measured the binding Kds between G5-or G13-psDiMan100% and labelled DC-SIGN under 3 different temperatures. Both of their quenching efficiencies (QEs) were found to decrease with the increasing temperature, indicating weakened interactions (larger Kds values). We then applied the Van't Hoff analysis to derive their binding thermodynamics by combining the two Gibbs free energy equations (eqs. 3 and 4). The changes of the standard binding enthalpy (ΔH 0 ) and entropy (ΔS 0 ) were obtained by taking a linear fit of the resulting ln(Kd) against the reciprocal of temperature, 1/T, via equation 5 (Fig. ). The results obtained from the fittings were summarized in Table .