We have investigated the consequences of introducing a bisecting GlcNAc moiety into biantennary N-glycans. Computational analysis of glycan conformation with pro-longed simulation periods in vacuo and in a solvent box revealed two main effects: backfolding of thea1–6 arm and stacking of the bisectingGlcNAc and the neighboring Man/ GlcNAc residues of both syn-thesis produced the bisecting biantennary decasaccharide N-glycan and itsa2–3(6)-sialylated variants.