Binding kinetics of liposome conjugated E-selectin and P-selectin glycoprotein ligand-1 measured with atomic force microscopy

[Display omitted] •Unbinding forces of PSGL-1/E-selectin and PSGL-1/E-selectin liposome were measured with AFM.•Binding kinetics of the two binding pairs were derived from measured unbinding forces.•Compared to E-selectin, E-selectin liposome has much higher association rate constant to PSGL-1.•E-selectin on liposome has much higher binding affinity to PSGL-1 than E-selectin. Various ligand-functionalized liposomes have been developed for targeted therapies. Typically, the binding properties of the ligands and targeted proteins are measured with surface plasmon resonance (SPR), where the proteins are immobilized on a rigid surface. However, the difference of protein-ligand binding kinetics between liposome-conjugated protein and rigid surface-conjugated protein is not fully understood. In this work, the binding kinetics of P-selectin glycoprotein ligand-1 (PSGL-1) and E-selectin conjugated on liposome and on rigid surfaces are investigated with Atomic Force Microscopy (AFM). The results suggest that protein orientation and diffusion on liposomal membrane can alter the binding kinetics of the protein-ligand interaction. Specifically, the association and dissociation rate constant of AFM probe-conjugated E-selectin and glass-conjugated PSGL-1 are measured as 9.32 × 104 M−1s−1 and 1.54 s−1, respectively. While for the liposome-conjugated E-selectin and glass-conjugated PSGL-1, the kinetic constants are measured as 5.00 × 107 M−1s-1 and 2.76 s−1, respectively. Thus, there is an order’s magnitude increase of binding affinity (from kd = 16.51 μM to kd = 0.06 μM) when protein is attached to liposome compared to attached to a rigid surface. The results might provide better understanding and pave the way for the future design of the ligand-targeted liposomes.