*2.3. Physicochemical Characterization of Nanoparticles*

All prepared NPs were examined in transmission electron microscope (TEM) (see Section 2.6). Optical extinction spectra were recorded on a Clariostar plate fluorimeter (BMG, Labtech Ortenberg, Germany) in the range of 400–800 nm according to manufacturer's instructions.

Hydrodynamic characteristics of the NPs were evaluated by method of photon correlation spectroscopy on a Malvern Zetasizer Nano-ZS instrument (Malvern Instruments, Malvern, UK). The measurements were performed at least 5 times for each sample.

All prepared NPs were subjected to agarose gel electrophoresis. Samples containing 5 μL (0.5 pmol) of each kind of NPs and 1 μL glycerol/water (1:1, v/v) were loaded into the wells of 0.8% agarose (Lonza Rockland, ME, USA) in Tris-Glycine buffer (250 mM glycine, 25 mM Tris, pH 8.3). The electrophoresis was carried out for 30 min at 5 V cm−1. Images were scanned using Epson Perfection 4990 Photo scanner (Seiko Epson Corporation, Suwa, Japan).
