Zetasizer zen3600

The ABMs-P were washed by PBS and distilled water until no colors were observed, and then they were suspended in distilled water, using an ultrasonic bath. The BMs and the previous ABMs-P supernatant (15–20 μL) were dropped on cropper grids and examined by TEM. The zeta potential was measured by a Zetasizer (ZEN3600; Malvern Instruments Ltd, Malvern, UK). All assays were performed in triplicate. DTS version 5.00 for Windows (Malvern Instruments Ltd) was used for statistical analysis.

Zetasizer (ZEN 3600; Malvern Instruments, Malvern, UK) was used as PAT for the analysis of particle size, PDI, and zeta potential.
The prepared nanosuspension was diluted 10 times with Milli-Q water to produce the working concentration. The diluted nanosuspension (1 mL) was taken in a zeta cell, and the cell was fixed into the Zetasizer to record the zeta potential. Similarly, the diluted nanosuspension (2 mL) was placed in the transparent glass cell to record the particle size and PDI. Analysis of samples was carried out at 90º scattering angle. This experiment was performed at 25°C.27

Particle size, size distribution, and zeta potential of the nanocrystals were determined by dynamic light scattering technique using Malvern Zetasizer (ZEN 3600; Malvern Instruments, Malvern, UK) and these were also as a part of process analytical tool. The analysis was carried out in triplicates at 25°C±1°C.38 ,39 (link) The results are presented as mean ± SD in Table 4.

BMs/DP/siRNA nanocomplexes were synthesized as shown in Figure 1. The nanocomplexes with different DP/siRNA weight ratios (ratio was set as 0, 0.13, 0.26, 0.52, 1.03, 1.29, 2.06, and 2.58) were prepared by adding appropriate amount of DP into siRNA-dispersed diethyl pyrocarbonate (DEPC) water, following that a fixed amount of BMs was added (BMs/siRNA weight ratio was 1:2). The mixture was vortexed for 2 min and incubated for 25 min at room temperature and collected. The formation of composites was determined by the gel electrophoresis assay. Briefly, the composites were added to 0.8% (w/v) agarose gel containing 1% (v/v) gel stain in tris acetate EDTA (TAE) buffer, the gel was run at 70 V for 20 min and imaged under UV transilluminator and a digital imaging system. The particle size and zeta potential of BMs/DP/siRNA were characterized by Zetasizer (ZEN3600; Malvern Instruments, Malvern, UK) in DEPC water.
To investigate whether the concentration of BMs affected the eventual formulation, BMs were treated with the DP/siRNA (DP/siRNA =2.06) complexes in DEPC water and incubated for 25 min (BMs/siRNA weight ratio was set as 1:5, 1:2, 1:1, 2:1, and 5:1). The products were characterized by agarose gel retardation assay and Zetasizer (ZEN3600).