Kinetics of pore formation in stearoyl-oleoyl-phosphatidylcholine vesicles by pH sensitive cell penetrating peptide GALA.

In order to engineer endosomal escape of drug carrying liposomes into the cytoplasm of target cells, the kinetics of bilayer poration by cell penetrating peptides needs to be well understood. To this end, we have studied pH-dependent pore formation in stearoyl-oleoyl-phosphatidylcholine vesicles as a function of concentration of the peptide GALA. Using laser scanning confocal microscopy, we measured the rate of fluorophore transport from the suspending medium into giant unilamellar vesicles across bilayer pores induced by GALA under acidic pH conditions. We also measured the mean pore size of GALA-induced pores in large unilamellar vesicles by electron microscopy. We fitted a mathematical model of pore formation kinetics to the measured rate of fluorophore transport across the giant vesicle bilayer to estimate the rate of pore formation as a function of GALA concentration. We observed that the number of pores per vesicle and the pore density increased with increasing GALA concentration.