Real-time monitoring of protein–liposome interaction kinetics using absorption, polarized intrinsic emission, and scattering (APIES): insights into protein corona formation

, (decreased by ∼20% at 25 mM compared to <5% at 100-150 mM after 30 minutes) hypsochromic spectral shifts (5.9 ± 1.2 nm at 25, 3.0 ± 0.8 nm at 150 mM after 3 hours). DLS confirmed that HSA-DMPC size decreased significantly within 200 seconds after mixing, and the gradual size increase afterwards. NTA confirmed the effect of ionic strength on HSA-liposome complex size and PSD. In conclusion, APIES is a powerful, yet simple tool for monitoring protein corona formation in real time (~1 s resolution), providing more detailed mechanistic insights beyond PSD changes.