Moreover, the UV-vis spectra of Gx-DiMan overlaid well with those of their parent GNPs with no significant red-shift and broadening of the SPR peaks, confirming no aggregation had taken place. The average number of glycan ligands on each GNP was estimated as ∼2200 ± 170 and ∼6290 ± 440 for G13-DiMan and G27-DiMan, respectively, by measuring the ligand amount difference between that added and that remained unbound in the supernatant after GNP conjugation using the phenol sulphuric acid carbohydrate quantitation as described previously. The average inter-glycan distances (d) were estimated as ∼0.93 and ∼0.80 nm for G13-DiMan and G27-DiMan, respectively, by using the Dh values and glycan valencies of Gx-DiMan as described previously (see SI, Table ).