As shown in Figure (Table for details), ΔG of wild type NTD is higher than that of R246I mutant in 8 systems except for 4-18 (PDB ID: 7L2E), DH1052 (PDB ID: 7LAB), S2X333 (PDB ID: 7LXY), suggesting that R246I mutant has the potential to impact the effectiveness of these 8 antibodies. As was expected, the energy contribution changes of R246 and I246 to the overall binding energy show the same tendency as ΔG (Figure ). Moreover, we performed 500ns MD simulation for 7L2F complex, whose trends of ΔΔG in different length of simulation time are consistent with the result of 30-60ns (Table ). This demonstrated that 60ns simulation is enough to obtain the variation tendency. Recent studies show that R246I can weaken the affinity of some antibodies, to varying degrees, including 5-24 (PDB ID:7L2F), 4A8 (PDB ID: 7C2L), 2-17 (PDB ID: 7LQW), FC05 (PDB ID: 7CWS) , and our predictions of percentage change in the relative binding free energy for above four systems are -53.76%, -40.58%, -13.92%, -19.13% respectively (Figure , S4). Therefore, our predicted trends of the relative binding free energy percentage are in good agreement with existing experimental findings, proving that our method is reliable to a certain extent.