Chitosan, Chitosan/IgG-Loaded, and N-Trimethyl Chitosan Chloride Nanoparticles as Potential Adjuvant and Carrier-Delivery Systems

Abstract

This work proposes a feasible, reproducible, and low-cost modified method to manufacture chitosan, chitosan/IgG-protein-loaded, and trimethylated chitosan nanoparticles, using microfluidics combined with the microemulsion technique, which differs from the traditional batch process of chitosan-based nanoparticles. The synthesis process consists of generating microreactors of chitosan-based polymer in a poly-dimethylsiloxane ψ-shaped microfluidic device and then crosslinking with sodium tripolyphosphate outside the cell. Transmission electron microscopy demonstrates an improvement in size control and distribution of the solid-shape chitosan nanoparticles (~80 nm) compared to the batch synthesis. Regarding chitosan/IgG-protein-loaded nanoparticles, these presented a core-shell morphology having a diameter of close to 15 nm. Raman and X-ray photoelectron spectroscopies confirmed the ionic crosslinking between the amino groups of chitosan and the phosphate groups of sodium tripolyphosphate in the fabricated samples and the total encapsulation of IgG protein during the fabrication of chitosan/IgG-loaded nanoparticles. Then, an ionic crosslinking and nucleation-diffusion process of chitosan-sodium tripolyphosphate was carried out during the nanoparticle formation, with and without IgG protein loading. The use of N-trimethyl chloride chitosan nanoparticles in vitro on human-keratinocyte-derived cell line HaCaT did not show side effects independently of its concentration from 1 to 10 μg/mL. Therefore, the proposed materials could be used as potential carrier-delivery systems.

Surface Modification of Polystyrene with Boronic Acid for Immunoaffinity-Based Cell Enrichment

“…An appreciable enhancement (54%, Figure ) of nitrogen concentration in PS-APBA-Ab versus PS-APBA indicates the presence of nitrogen from IgG, as IgGs are proteins and are enriched with amino (NH 2 ) groups. In addition, the ratio of nitrogen to carbon concentration ([N]/[C]) is increased 74% (from 0.12 to 0.21%) in PS-APBA-Ab surface (Figure ) which supports the presence of protein on the surface after antibody immobilization. , Another indicator of Ab existence on the surface is sulfur which is expected due to the disulfide bridge of Ab structure. , The concentration of sulfur at 0.3% is slightly less than the approximate sulfur content in an antibody (i.e., 0.45%) and just above the detection limit of XPS (about 0.1%) . This suggests a high-density layer of Abs.…”

“…In the PS-APBA-Ab graph, another signal at 164 eV is seen that matches sulfur (Figure 6g). This peak shows disulfide bridge (S�S) which can indicate the existence of Ab on the surface, 64,65 while no similar signal is detected on PS-APBA and PS-Bare samples.…”

“…67,68 Another indicator of Ab existence on the surface is sulfur which is expected due to the disulfide bridge of Ab structure. 64,65 The concentration of sulfur at 0.3% is slightly less than the approximate sulfur content in an antibody (i.e., 0.45%) and just above the detection limit of XPS (about 0.1%). 69 This suggests a high-density layer of Abs.…”