The encapsulation of CAT-Aco in DGL NPs was examined using an acrylamide gel shift assay, with naked CAT-Aco as the control. NPs composed of different weight ratios of PGP-PEG-DGL were mixed with appropriate amounts of 2× loading buffer and then electrophoresed on an 8.0% acrylamide gel containing 5 mM Tris and 50 mM glycine, at pH 8.3, under non-denaturing conditions (in the absence of SDS) to preserve the binding. The protein bands were stained with rabbit polyclonal anti-CAT antibody (Ab 6573, Abcam; 1:1000) and visualized with Immobilon Western Chemiluminescent HRP substrate (Millipore).
cl PGP-PEG-DGL/CAT-Aco NPs were freshly prepared with a specified ratio (2.5:1, DGL to CAT-Aco, w/w). The mean diameter and zeta potential of the NPs were determined with a Zeta Potential/Particle Sizer Nicomp 380 ZLS system (PPS Nicomp Particle Size System, USA). The morphology of the cl PGP-PEG-DGL/CAT-Aco NPs was examined under a high-resolution transmission electron microscope (TEM) (JEM-2010HT, Japan).
cl PGP-PEG-DGL/CAT-Aco NPs were freshly prepared with a specified ratio (2.5:1, DGL to CAT-Aco, w/w). The mean diameter and zeta potential of the NPs were determined with a Zeta Potential/Particle Sizer Nicomp 380 ZLS system (PPS Nicomp Particle Size System, USA). The morphology of the cl PGP-PEG-DGL/CAT-Aco NPs was examined under a high-resolution transmission electron microscope (TEM) (JEM-2010HT, Japan).