Protonation states were assessed using H++ 3.2 at pH 7.4. A cubic explicit water box described using the TIP3P model was used to solvated the complex system, which was extended by 12 Å from the solute. An atmosphere of 150 mM NaCl was also included in all simulations. The generated models were parametrized using CHARMM36 all-atom additive force fields for protein and glycans . 8,000 steps of minimization including 6,000 steps of steepest descent minimization and 2,000 steps of conjugate gradient minimization were performed to remove bad contacts during the energy minimization phase. Subsequently, the temperature was incrementally changed from 0 to 300 K for 125ps at 1 fs/step. Equilibration in NPT ensemble was run at 1.0 bar and 300 K for 500,000 steps at 2fs/step. Pmemd program implement in Amber18 software package was used to run the minimization, heating simulations with position constraints (1 kcal/mol/Å 2 ) on protein and dihedral Angle constraints on carbohydrates.