A New Approach to Antivenom Preparation Using Chitosan Nanoparticles Containing Echis Carinatus Venom as A Novel Antigen Delivery System

Document Type: Research article


1 Department of Nanotechnology, Faculty of Science, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.

2 Department of Human Vaccines and Serum, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Karaj, Iran.

3 Department of Research and Development, Hakim Pharmaceutical Company, Tehran, Iran

4 Department of Nanotechnology, Tehran University of Medical Science, Tehran, Iran


In recent years, use of biodegradable polymers based nanoparticles has received high
interest in the development of vaccines delivery vehicles. The aim of study was to prepare
chitosan nanoparticles (CS NPs) for loading Echis carinatus (EC) venom and evaluate their
potential as an adjuvant and antigen delivery system on a pilot scale. CS NPs were prepared
using ionic gelation method, and their characteristics were optimized. Venom-loaded CS NPs
prepared under optimum conditions and traditional venom-loaded adjuvants were used to
hyperimmunization of horse. Under optimum conditions, particle size, polydispersity index
(PDI), and zeta potential of CS NPs were 127.9 ± 15 nm, 0.29, and +19.8 ± 1.92 mV, while
those of venom-loaded CS NPs were 182.4 ± 20 nm, 0.35, +26.8 ± 1.98 mv, respectively. All
CS NPs had integrated surface and good morphology. Optimum loading concentration of EC
venom was 500 μg/mL. The loading capacity (LC) and loading efficiency (LE) were 87% and
94%, respectively, and release profile of venom-loaded CS NPs showed suitable correlation
with Higuchi kinetics. Stability test showed good stability of the venom encapsulated in CS
NPs. Furthermore, antivenom plasma obtained using the new antigen delivery system had
significantly higher potency (P < 0.05) for neutralizing the venom than that obtained using
conventional system. These results suggested that venom-loaded CS NPs could be a suitable
alternative to conventional adjuvant for development antivenom.


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