Figure 2.

Dual modal click-functionalization of liposomes enables conjugation of varied endothelial adhesion molecule targeting species. (a,b) Gel exclusion chromatography elution data of (a) DBCO liposomes incubated with ¹²⁵I-azide functionalized mAb (green line) or scFv (red line) and (b) azide liposomes, as labeled in (a). The early elution peak (5–7 mL) in both graphs signals the radioactive ligand bound to the liposome, and the later peaks (max peak 14 mL for mAb, 17.5 mL for scFv) signals the unbound ligand elution signal. The y-axis # ligand bound calculated from a known addition of ligands, with scFv azide/DBCO species added 2.0× greater than mAb DBCO/azide. Area under the curve analyses of elution graphs calculate 85–95% conjugation efficiency. (c,d) Kinetics data of azide/DBCO modified IgG conjugation reaction to DBCO/azide functionalized liposomes. ¹²⁵I labeled IgG-azide/DBCO traced as in a,b. (e) Comparison of bound ligand over time by click species orientation. (f) Extent of bound ligands, particle size, and PDI of kinetics data samples shown in c,d.