*2.4. Dynamic Light Scattering (DLS) and Electrophoretic Light Scattering (ELS)*

DLS measurements were performed utilizing a Nano ZS (Malvern Panalytical, Malvern, United Kingdom) with a laser wavelength of λ = 633 nm with non-invasive back-scatter (NIBS) technology [22]. The particle size is reported as the hydrodynamic diameter (dH). The particle size distribution (PDI) was measured using pure water as a dispersant with a refractive index RI of 1330 and a viscosity of 0.8872 cP at 25 ◦C. Samples obtained from the automated HT-nanoprecipitation were measured at 25 ◦C in a micro cuvette (Brand GmbH, Wertheim, Germany) without any filtering step with the following settings: measurements of each sample were repeated three times for 10 sec at 25 ◦C. The samples obtained from batch nanoprecipitation were measured at a dilution of 1:10 up to 1:100 utilizing the following settings: five repeated measurements, each with five runs of 30 s. The zeta-potential of the lyophilized NPs was investigated by ELS using the same instrument at 25 ◦C with three repeated measurements.

The apparent degradation behavior of the NPs was analyzed by DLS by monitoring changes in the mean count rate at fixed measurement settings: measuring position at 4.65, attenuator factor 7 at 37 ◦C [21]. Before investigating, NPs were mixed with the enzyme solution (a lipase from Candida rugosa) in a 1:4 mass ratio of polymer to enzyme and incubated at 37 ◦C for pre-determined timepoints.
