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Carboxymethyl-beta-glucan/chitosan nanoparticles for antigen delivery

Various polysaccharides such as chitosan and β-glucan possess immunological properties that are advantageous for vaccine adjuvants. Therefore, in the presented study, chitosan and carboxymethyl-β-glucan were combined as nanoparticles, coupled to the model antigen ovalbumin as vaccine adjuvant and investigated in vivo in a mouse model.


Cordeiro AS, Farsakoglu Y, Crecente-Campo J, de la Fuente M, González SF, Alonso MJ. Carboxymethyl-β-glucan/chitosan nanoparticles: new thermostable and efficient carriers for antigen delivery. Drug Deliv Transl Res. 2021 Aug;11(4):1689-1702. doi: 10.1007/s13346-021-00968-9. Epub 2021 Apr 1. PMID: 33797035; PMCID: PMC8015750.

In recent decades, vaccine development has focused mainly on the development of safe antigens and more efficient adjuvants. Adjuvants are needed to enhance the body's immune response to the vaccine. For example, the RNA or DNA vaccines used in the Covid-19 pandemic require delivery systems to present the genetic antigenic material to immune cells. Currently, vaccines primarily use aluminum (alum) or lipid-based adjuvants, or a combination of both.
An alternative may be particle-based adjuvants that deliver antigens via lipid-based micro- and nanoparticles. These have the advantages of providing protection against degradation of the antigens, controlled transmission, and enhanced recognition of the antigens via antigen-presenting cells. In addition, carriers can also be designed to include adjuvants in their structure to enhance the immune response.
Enhanced immune cell recognition can be achieved by taking advantage of natural polysaccharides such as β-glucan and chitosan. Due to the presence of similar structural patterns as to bacteria and viruses, these can enhance the recognition of the vaccine by immune cells. For this reason, the study presented here worked on the development of carboxylmethyl-β-glucan/chitosan nanoparticles and investigated their antigen delivery ability. For this purpose, the model antigen ovalbumin (OVA) was bound to the generated nanoparticles and injected into mice in vivo to study the interaction with immune cells. In addition, the immune response of the OVA-NP was analyzed in the mouse model. The chitosan used for this purpose was GMP chitosan HCl with a degree of deacetylation of 80-95% and a molecular weight of 47 kDa from Heppe Medical Chitosan GmbH.


  • The amount of chitosan affected particle size, polydispersion index (PdI) and zeta potential, only particles with an excess of chitosan (4:1, 2:1) had a PdI˂0.2, a small size and positive charges
  • Production yield of 85%→ both polysaccharides have been successfully integrated into nanoparticles
  • CS:CMβG 2:1 was successfully coupled with up to 6.7% OVA (were used for the further experiments), the OVA-NPs exhibited a size of 160-170 nm, a zeta potential of 30 mV, and a spherical structure
  • Evidence via Western blot that the antigen was not affected via binding with the NPs
  • Nanoparticles were stable for 2 months at 4 °C and showed no change in antigen activity after freeze-drying and subsequent storage at 40 °C and 75% humidity for 28 days
  • In vivo: uncharged and charged OVA-NP reached lymph nodes and activated dendritic cells (DCs)
  • No differences in T-cell activation, interaction with antigen-presenting cells, and antibody levels were observed when compared with free OVA, OVA-NP, and alum-OVA
  • Higher immune response in antibody-secreting cells for OVA-NP compared to free OVA

Summary: In the presented study, chitosan and carboxylmethyl-β-glucan as two polysaccharides with immunological properties were combined as nanoparticles and successfully coupled with OVA. These could successfully activate lymph nodes, dendritic cells and T cells in vivo. In addition, it was shown that the nanoparticles remained stable upon freeze-drying and subsequent storage for 28 days at 40 °C, which is advantageous for subsequent storage and distribution of the vaccine. Link to article:


chitosan, nanoparticles, vaccine

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